Using Goat’s Milk,Barley Flour,Honey, and Probiotic to Manufacture of Functional Dairy Product

Stirred yogurt manufactured using probiotic culture which usually called Rayeb milk in the Middle East region is one of the most important functional fermented milk products. To increase the health and functionality properties to this product, some ingredients like fruits, cereal, and whey protein are used in production. This study was carried out to prepare functional Rayeb milk from goat’s milk, barley flour (15%) and honey (4%) mixtures using ABT culture. Also, vanilla and cocoa powder were used as flavorings. Adding barley flour and honey to goat’s milk increased curd tension and water-holding capacity and decreased coagulation time and susceptibility to syneresis. The values of carbohydrate, total solids, dietary fiber, ash, total protein, water soluble nitrogen, total volatile fatty acids, unsaturated fatty acids, oleic, linoleic, α-linolenic acids, and antioxidant activity were higher in Rayeb milk supplemented with barley flour and honey than control. The viabilities of Lactobacillus acidophilus and Bifidobacterium lactis Bb12 (Chr. Hansen’s Lab A/S) increased in fortified Rayeb milk. The recommended level of 10⁷ cfu g^?1 of bifidobacteria as a probiotic was exceeded for these samples. Addition of vanilla (0.1%) or cocoa powder (0.5%) improved the sensory properties of fortified Rayeb milk.     

Sorption of Acid Blue 9 Using Hydrilla verticillata Biomass—Optimization,Equilibrium, and Kinetics Studies

In the present study, the parameters, temperature, adsorbent dose, contact time, adsorbent size, and agitation speed were optimized for Acid Blue 9 removal from aqueous medium by Hydrilla verticillata biomass using response surface methodology (RSM). The optimum conditions for maximum removal of Acid Blue 9 from an aqueous solution of 100 mg/L were as follows: temperature 30.63°C, adsorbent dose 2.88 g/L, contact time 180 min, adsorbent size 120 mesh (0.124 mm), and agitation speed 237.39 rpm. At these optimized conditions, batch adsorption experiments were conducted to study the effect of pH and initial dye concentration for the removal of Acid Blue 9 dye. The optimum initial pH and initial dye concentration values for Acid Blue 9 removal were found to be 3.0 and 100 mg/L, respectively. Kinetic and equilibrium studies were carried out for the experimental results. From the kinetic studies it was found that pseudo-second-order kinetics suits the system well. From the equilibrium studies, the Freundlich isotherm fits the data well.     

OPEN HEART OPERATIONS—Results in 100 Consecutive Cases,Using Extracorporeal Circulation

In one hundred consecutive cases in which operations were done on the heart with the surgical field exposed to view and circulation was shunted to extracorporeal apparatus, there were six deaths. None were attributable to the by-pass procedure. The lesions were of various kinds, congenital and acquired. In one case a fibromyxosarcoma that was obstructing the right ventricular outflow tract and invading the main pulmonary artery was successfully excised.     

AORTIC STENOSIS—Surgical Treatment Under Direct Vision,Using the Heart-Lung Machine

Cardiac operation with the operative field under direct vision and the patient''s oxygenation maintained by a heart-lung machine affords an excellent means of treating aortic stenosis, for it allows unlimited time in which to repair aortic stenosis no matter how involved the cusps are with calcium. The valves can be sculptured and the commissures cut at their place of fusion. Concomitant aortic insufficiency can also be taken care of at the same time, as can associated mitral valve stenosis and insufficiency.This method was used in ten patients. Two died. Eight were considerably improved.     

Reconstituted Micelle Formation Using Reduced, Carboxymethylated Bovine κ-Casein and Human β-Casein

In milk, κ-casein, a mixture of disulfide-bonded polymers, stabilizes and regulates the size of the unique colloidal complex of protein, Ca²⁺ and inorganic phosphate (P_i) termed the casein (CN) micelle. However, reduced, carboxymethylated bovine κ-CN (RCM-κ) forms fibrils at 37°C and its micelle-forming ability is in question. Here, the doubly- and quadruply-phosphorylated human β-CN forms and 1:1 (wt:wt) mixtures were combined with RCM-κ at different β/κ weight ratios. Turbidity (OD_{400 nm}) and a lack of precipitation up to 37°C were used as an index of micelle formation. Studies were with 0, 5 and 10 mM Ca²⁺ and 4 and 8 mM P_i. The RCM-κ does form concentration-dependent micelles. Also, β-CN phosphorylation level influences micelle formation. Complexes were low-temperature reversible and RCM-κ fibrils were seen. There appears to be equilibrium between fibrillar and soluble forms since the solution still stabilized after fibril removal. The RCM-κ stabilized better than native bovine κ-CN.     

Simple Detection of Xylosidase Activity in Single Colonies,Using 1-Naphthyl-β-d-xylopyranoside

Since the xylosidase of Bacillus pumilus hydrolyzed 1-naphthyl-β-d-xylopyranoside (naphthyl-X) to produce xylose and 1-naphthol and a chromogenic azo compound is produced by coupling 1-naphthol and Fast Blue Salt B, a simple method for detection of xylosidase activity in single colonies was studied. Escherichia coli JM109 carrying the xylosidase gene of B. pumilus was cultivated at 37°C for 18 h on an LB plate containing 0.5 mg/ml naphthyl-X, and then the plate was overlaid with 3 ml of a top layer containing 24 mg of agar and 6 mg of Fast Blue Salt B. After incubation of the plate at 37°C for 1 h, each colony became reddish-brown. Even a small colony with the xylosidase on the plate was easily distinguished from colonies without the enzyme.     

Enhancing Proper Sitting Position Using a New sEMG Protocol, The “Minimax” Procedure, With Boolean Logic

The modification of posture in children using sEMG is a new area that offers much potential to aid disabled children regain function. This article describes a new protocol, the minimax procedure, to aid children regain motor function in sitting. The protocol uses selective reinforcement to encourage recruiting/relaxing target sites (gluteus medius/maximus) in a predetermined way so as to encourage good form in sitting. Children often are unaware how to make changes in posture, and when they do succeed the progress is so slow that sustaining motivation is a challenge. This paper describes a procedure that helps the child recognize when they have achieved the correct posture, immediately rewards them for doing so, and generates data using Boolean operations to numerically substantiate the clinically observed changes in posture.     

Dissecting Virus Entry: Replication-Independent Analysis of Virus Binding,Internalization, and Penetration Using Minimal Complementation of β-Galactosidase

Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this key process in virus infection. Here we describe novel, highly adaptable viral entry assays making use of minimal complementation of the E. coli β-galactosidase in mammalian cells. Enzyme activity is reconstituted when a small intravirion peptide (α-peptide) is complementing the inactive mutant form ΔM15 of β-galactosidase. The method allows to dissect and to independently detect binding, internalization, and fusion of viruses during host cell entry. Here we use it to confirm and extend current knowledge on the entry process of two enveloped viruses: vesicular stomatitis virus (VSV) and murine hepatitis coronavirus (MHV).     

Using codon optimization, chaperone co-expression, and rational mutagenesis for production and NMR assignments of human eIF2α

Producing a well behaved sample at high concentration is one of the main hurdles when starting a new project on an interesting protein. Especially when one attempts to overexpress a eukaryotic protein in bacteria, some difficulties are encountered, such as low expression level, low solubility, or even lack of folded structure. Overexpression in prokaryotic systems is highly desirable for cost-effective production of different isotope-labeled samples needed for NMR studies. Here we describe generally applicable methods for obtaining highly concentrated protein samples efficiently. This approach was developed as we tried to produce a NMR-suitable sample of the 35 kDa human translation initiation factor eIF2 alpha, a protein that expresses poorly in E. coli and has very low solubility. First, an E. coli codon-optimized gene was synthesized on a thermal cycler, which increased the expression level by a factor of two. Second, we used co-expression of bacterial chaperone proteins, which largely increased the fraction of correctly folded protein found in the soluble phase. Third, we used rational mutagenesis guided by both the sequence alignment among homologues and the homology of one domain to a known fold for improving solubility and stability of the target protein by tenfold. Combining all these methods made it possible to produce from a one-liter preparation a 0.5 mM sample of human eIF2 alpha that showed well-resolved NMR spectra and enabled nearly complete assignment of the protein. These methods may be generally useful for studies of other eukaryotic proteins that are otherwise difficult to express and exhibit poor solubility.     

Using Multiconformation Continuum Electrostatics to Compare Chloride Binding Motifs in α-Amylase, Human Serum Albumin, and Omp32

Ions are a ubiquitous component of the cellular environment, transferring into cells through membrane-embedded proteins. Ions bind to proteins to regulate their charge and function. Here, using multiconformation continuum electrostatics (MCCE), we show that the changes of chloride binding to α-amylase, human serum albumin (HSA) and Omp32 with pH, and of α-amylase with mutation agree well with experimental data. The three proteins represent three different types of binding. In α-amylase, chloride is bound in a specific buried site. Chloride binding is strongly coupled to the protonation state of a nearby lysine. MCCE calculates an 11-fold change in chloride affinity between the wild-type α-amylase and the K300R mutant, in good agreement with the measured 10-fold change. Without considering the coupled protonation reaction, the calculated affinity change would be more than 10⁶-fold. In HSA, chlorides are distributed on the protein surface. Although HSA has a negative net charge, it binds more anions than cations. There are no highly occupied binding sites in HSA. Rather, there are many partially occupied sites near clusters of basic residues. The relative affinity of bound ions of different charges is shown to depend on the distribution of charged residues on the surface rather than the overall net charge of the protein. The calculated strong pH dependence of the number of chlorides bound and the anion selectivity agree with those of previous experiments. In Omp32, chlorides are stabilized in an anion-selective transmembrane channel in a pH-independent manner. The positive electrostatic potential in Omp32 results in about two chlorides and no cations bound in the transmembrane region of this anion-selective channel. The studies here show that with the ability to sample multiple binding sites and coupled protein protonation states, MCCE provides a powerful tool to analyze and predict ion binding. The calculations overestimate the affinity of surface chloride in HSA and Omp32 relative to the buried ion in amylase. Differences between ion-solvent interactions for buried and surface ions will be discussed.     

Reusable Solid-Phase Supports for Oligonucleotide Synthesis Using Hydroquinone-O,O'-diacetic Acid (“Q-Linker”)

Abstract

Reusable solid-phase supports for large scale oligonucleotide synthesis have been prepared by converting amino derivatized supports into hydroxyl supports. Rapid nucleo side attachment, via a Q-linker arm, was automatically performed on the DNA synthesizer using HBTU and DMAP as the coupling reagents. All steps were suitable for automation and ～ 1.5 h was required to prepare the supports for reuse. Up to twelve consecutive syntheses of a 20-mer phosphorothioate were performed on a synthesis column.     

Using functional–structural plant models to study, understand and integrate plant development and ecophysiology

Functional–structural plant models (FSPMs) explore and integrate relationships between a plant’s structure and processes that underlie its growth and development. In recent years, the range of topics being addressed by scientists interested in functional–structural plant modelling has expanded greatly. FSPM techniques are now being used to dynamically simulate growth and development occurring at the microscopic scale involving cell division in plant meristems to the macroscopic scales of whole plants and plant communities. The plant types studied also cover a broad spectrum from algae to trees. FSPM is highly interdisciplinary and involves scientists with backgrounds in plant physiology, plant anatomy, plant morphology, mathematics, computer science, cellular biology, ecology and agronomy. This special issue of Annals of Botany features selected papers that provide examples of comprehensive functional–structural models, models of key processes such as partitioning of resources, software for modelling plants and plant environments, data acquisition and processing techniques and applications of functional–structural plant models for agronomic purposes.     

Spatiotemporal Analysis of Infant Measles Using Population Attributable Risk in Shandong Province, 1999–2008

Background

Reduction of measles incidence and mortality has been encouraging in China. However, it remains an important public health concern among infants. This study aimed to examine the space–time distribution pattern of infant measles occurrence for the period of 1999–2008 in Shandong, China.

Methods and Findings

Measles cases among infants aged younger than 1 year were obtained from the national infectious diseases reporting information system. A spatiotemporal analysis using population attributable risk percent (PAR%) was used to distinguish between multiple geographic clusters of potential interest. The analysis detected 29 statistically significant space–time clusters with the most likely cluster in Zaozhuang City from 2006 to 2008. Of the 28 secondary clusters, 22 were found in 2008. The map of PAR%, relative risk (RR) and space–time cluster analysis indicated that the clusters were generally unchanged, and were found south-west and north-west of Shandong. The Lanshan District in Linyi had the highest PAR%, while highest RR was in the Yicheng District in Zaozhuang.

Conclusion

There were significant space-time clusters of infant measles in Shandong over the study period. PAR% is an effective way to analyze multiple clusters from their application like RR. Interrupting measles circulation and maintaining routine coverage over 95% may be the only effective strategy to achieve measles elimination.     

Using DNA barcodes for assessing diversity in the?family Hybotidae (Diptera,Empidoidea)

Empidoidea is one of the largest extant lineages of flies, but phylogenetic relationships among species of this group are poorly investigated and global diversity remains scarcely assessed. In this context, one of the most enigmatic empidoid families is Hybotidae. Within the framework of a pilot study, we barcoded 339 specimens of Old World hybotids belonging to 164 species and 22 genera (plus two Empis as outgroups) and attempted to evaluate whether patterns of intra- and interspecific divergences match the current taxonomy. We used a large sampling of diverse Hybotidae. The material came from the Palaearctic (Belgium, France, Portugal and Russian Caucasus), the Afrotropic (Democratic Republic of the Congo) and the Oriental realms (Singapore and Thailand). Thereby, we optimized lab protocols for barcoding hybotids. Although DNA barcodes generally well distinguished recognized taxa, the study also revealed a number of unexpected phenomena: e.g., undescribed taxa found within morphologically very similar or identical specimens, especially when geographic distance was large; some morphologically distinct species showed no genetic divergence; or different pattern of intraspecific divergence between populations or closely related species. Using COI sequences and simple Neighbour-Joining tree reconstructions, the monophyly of many species- and genus-level taxa was well supported, but more inclusive taxonomical levels did not receive significant bootstrap support. We conclude that in hybotids DNA barcoding might be well used to identify species, when two main constraints are considered. First, incomplete barcoding libraries hinder efficient (correct) identification. Therefore, extra efforts are needed to increase the representation of hybotids in these databases. Second, the spatial scale of sampling has to be taken into account, and especially for widespread species or species complexes with unclear taxonomy, an integrative approach has to be used to clarify species boundaries and identities.     

Exposure-Based, ‘Like-for-Like’ Assessment of Road Safety by Travel Mode Using Routine Health Data

Background

Official reports on modal risk have not chosen appropriate numerators and denominators to enable like-for-like comparisons. We report age- and sex-specific deaths and injury rates from equivalent incidents in England by travel mode, distance travelled and time spent travelling.

Methods

Hospital admissions and deaths in England 2007–2009 were obtained for relevant ICD-10 external codes for pedestrians, cyclists, and car/van drivers, by age-group and sex. Distance travelled by age-group, sex and mode in England (National Travel Survey 2007–2009 data) was converted to time spent travelling using mean trip speeds. Fatality rates were compared with age-specific Netherlands data.

Results

All-age fatalities per million hours’ use (f/mhu) varied over the same factor-of-three range for both sexes (0.15–0.45 f/mhu by mode for men, 0.09–0.31 f/mhu for women). Risks were similar for men aged 21–49 y for all three modes and for female pedestrians and drivers aged 21–69 y. Most at risk were: males 17–20 y (1.3 f/mhu (95% CI 1.2–1.4)) for driving; males 70+ (2.2 f/mhu(1.6–3.0)) for cycling; and females 70+ (0.95 f/mhu (0.86–1.1)) for pedestrians. In general, fatality rates were substantially higher among males than females. Risks per hour for male drivers <30 y were similar or higher than for male cyclists; for males aged 17–20 y, the risk was higher for drivers (33/Bn km (30–36), 1.3 f/mhu (1.2–1.4)) than cyclists (20/Bn km (10–37), 0.24 f/mhu (0.12–0.45)) whether using distance or time. Similar age patterns occurred for cyclists and drivers in the Netherlands. Age-sex patterns for injuries resulting in hospital admission were similar for cyclists and pedestrians but lower for drivers.

Conclusions

When all relevant ICD-10 codes are used, fatalities by time spent travelling vary within similar ranges for walking, cycling and driving. Risks for drivers were highest in youth and fell with age, while for pedestrians and cyclists, risks increased with age. For the young, especially males, cycling is safer than driving.     

Antisense Knockdown of PKC-α Using LNA-Oligos

Abstract

Full-length and 4 nucleotides truncated Locked Nucleic Acid (LNA) modifications of ISIS 3521 were compared for antisense properties in a cellular assay. ISIS 3521 is a 20-mer phosphorothioate designed to hybridise to human protein kinase C-α (PKC-α) mRNA and is currently submitted to clinical trials against cancer. We report that LNA can potentate this antisense oligo and retain the antisense potential with shorter oligos.     

Using γ-emitting artificial radionuclides,released by nuclear plants,as markers of restricted movements by chub,Leuciscus cephalus,in a large river,the Lower Rhône

Synopsis On the basis that -emitting artificial radionuclides, present in nuclear plant radioactive wastes, can be considered as natural collective markers of fish living downstream from a radioactive discharge, we studied the restricted movements of chub in a reservoir on the Lower Rhône river where the Marcoule nuclear plant is located. A qualitative determination, based on the detection of specific radioelements in our samples originating from radioactive waste, and a quantitative determination of the cesium-137 concentration in the samples, were used to identify fish radioactively marked by the effluent. Individual measurements of -radioactivity in 49 adult chub captured at two stations, each 2 km long and 6 km apart, divided the fish into two distinct sub-units: one living downstream of the discharge pipe, in which 73% were marked, and the other upstream, in which 79% were not marked. Similar results were found in two neighbouring stations when we analysed previous radioecological measurements of -radioactivity in groups of chub. After combining all the data concerning chub, detailed information was obtained on the spatial stability of the chub population in the reservoir. Two spatially different stocks were found and each stock can be divided in two components: a sedentary component that remains in a restricted zone (its home range), and a mobile component that undertakes movements between the two zones.     

Using Dental Enamel Wrinkling to Define Sauropod Tooth Morphotypes from the Ca?adón Asfalto Formation,Patagonia, Argentina

The early Middle Jurassic is regarded as the period when sauropods diversified and became major components of the terrestrial ecosystems. Not many sites yield sauropod material of this time; however, both cranial and postcranial material of eusauropods have been found in the Cañadón Asfalto Formation (latest Early Jurassic–early Middle Jurassic) in Central Patagonia (Argentina), which may help to shed light on the early evolution of eusauropods. These eusauropod remains include teeth associated with cranial and mandibular material as well as isolated teeth found at different localities. In this study, an assemblage of sauropod teeth from the Cañadón Asfalto Formation found in four different localities in the area of Cerro Condor (Chubut, Argentina) is used as a mean of assessing sauropod species diversity at these sites. By using dental enamel wrinkling, primarily based on the shape and orientation of grooves and crests of this wrinkling, we define and describe three different morphotypes. With the exception of one taxon, for which no cranial material is currently known, these morphotypes match the local eusauropod diversity as assessed based on postcranial material. Morphotype I is tentatively assigned to Patagosaurus, whereas morphotypes II and III correspond to new taxa, which are also distinguished by associated postcranial material. This study thus shows that enamel wrinkling can be used as a tool in assessing sauropod diversity.     

Prediction of Radiation Esophagitis in Non–Small Cell Lung Cancer Using Clinical Factors,Dosimetric Parameters,and Pretreatment Cytokine Levels

Radiation esophagitis (RE) is a common adverse event associated with radiotherapy for non–small cell lung cancer (NSCLC). While plasma cytokine levels have been correlated with other forms of radiation-induced toxicity, their association with RE has been less well studied. We analyzed data from 126 patients treated on 4 prospective clinical trials. Logistic regression models based on combinations of dosimetric factors [maximum dose to 2 cubic cm (D2cc) and generalized equivalent uniform dose (gEUD)], clinical variables, and pretreatment plasma levels of 30 cytokines were developed. Cross-validated estimates of area under the receiver operating characteristic curve (AUC) and log likelihood were used to assess prediction accuracy. Dose-only models predicted grade 3 RE with AUC values of 0.750 (D2cc) and 0.727 (gEUD). Combining clinical factors with D2cc increased the AUC to 0.779. Incorporating pretreatment cytokine measurements, modeled as direct associations with RE and as potential interactions with the dose-esophagitis association, produced AUC values of 0.758 and 0.773, respectively. D2cc and gEUD correlated with grade 3 RE with odds ratios (ORs) of 1.094/Gy and 1.096/Gy, respectively. Female gender was associated with a higher risk of RE, with ORs of 1.09 and 1.112 in the D2cc and gEUD models, respectively. Older age was associated with decreased risk of RE, with ORs of 0.992/year and 0.991/year in the D2cc and gEUD models, respectively. Combining clinical with dosimetric factors but not pretreatment cytokine levels yielded improved prediction of grade 3 RE compared to prediction by dose alone. Such multifactorial modeling may prove useful in directing radiation treatment planning.