bluuebunny/crossref_metadata_2025 huggingface dataset:
Estimating the degree of heat treatment given to milk
Authors: WILBEY R ANDREW
Published: August 2007

A wide range of tests based on enzyme activity and chemical analyses have been developed. Enzyme based tests are appropriate for pasteurization and mild heat treatments, while the severe heat treatments associated with ultra high temperature and in‐container sterilization processes may be assessed by chemical analyses.
Measurement of mastitis in first lactation heifers and third to fourth lactation cows
Authors: BURKE J , CAMPBELL A C
Published: August 2007

The influence of lactation stage on susceptibility to mastitis infection in Friesian dairy cows was investigated. The cattle studied included third to fourth lactation cows and both summer and autumn calving groups of heifers. No increase in vulnerability to invasion by mastitis pathogens in recently calved cattle was demonstrated. Analysis of raw milk samples using ATP bioluminescence produced variation in results, suggesting that further work to develop the technique is required.
The development of a model butter reworker
Authors: GALLAT STEPHANIE , WILBEY R ANDREW , BELL ALAN E
Published: August 2007

The throughput of commercial butter reworkers is too great for experimental work. A laboratory scale reworker was designed and constructed to enable quantitative investigation of the reworking process.
Starter cultures employed in the manufacture of biofermented milks
Authors: Marshall V M , Tamime A Y
Published: August 2007

The market for fermented milks is expanding as more consumers become interested in the possible health aspects of the product. This paper reviews the types of bacteria that are currently being used in the manufacture of biofermented milks. The requirements of a starter culture in terms of ability to ferment milk to a stable product and to provide the necessary sensory properties for consumer demands are outlined. The paper also discusses and highlights some of the problems encountered in developing milks for the health market using probiotic organisms such as Lactobacillus acidophilus and species of Bifidobacterium.
What iceberg? Crisis management in action
Authors: Johnston Mike
Published: August 2007

No firm is exempt from the potentially devastating effects of a crisis. Examples of how two crises have been dealt with are provided, and lessons drawn from each. Handling a crisis is considered in three phases: pre‐crisis, crisis and post‐crisis. In the pre‐crisis phase emphasis is on prevention and preparation. Firms should establish a crisis management team and prepare a plan for dealing with crises. Some practical suggestions are provided for the crisis phases where both communication and company response are important. In the post‐crisis phase firms need to take stock and go on the offensive to regain lost ground.
Chemical and microbial stability of fluid milk in response to packaging and dispensing
Authors: Erickson Marilyn C
Published: August 2007

Chemical and microbiological stability of milk packaged in one litre pouches, a two litre paperboard box (with plastic liner) or a one gallon plastic jug were compared. The cardboard box gave the greatest protection against light induced lipid oxidation while significantly higher microbial populations were found in the larger containers (gallon jug and paperboard box) than in the smaller containers (one litre pouches) after one week of storage. Microbial populations were also greater in milk whose package had been opened and dispensed from previously than from milk found in an unopened package. These microbial data would suggest that one gallon of milk would remain fresher for the consumer when packaged in multi‐unit packages than if packaged in one single container.
Preparation and properties of ice cream type frozen yogurt
Authors: INOUE K , SHIOTA K , ITO T
Published: August 2007

Changes in sensory ratings and chemical properties of ice cream type frozen yogurt (fat content >8.0% with varying pH values) were investigated during storage for 6 months at –35°C. No appreciable change was observed in the structure, acidity and pH values of the products during storage. In addition, there was no increase in thiobarbituric acid values of the products during storage. Viable lactic acid bacteria decreased in number with increasing storage period. Different sensory ratings were obtained for assessors who were familiar with yogurt and for assessors who were not. The product having a pH value of 5.5 was the most preferred ice cream type frozen yogurt in the products examined.
Characteristics of fried paneer prepared from mixtures of buffalo milk, skimmed milk, soy milk and buttermilk
Authors: SHARMA H K , SINGHAL R S , KULKARNI P R
Published: August 2007

Paneer was prepared by substituting 3, 5 and 10% of buffalo milk with either soy milk, buttermilk or skimmed milk prepared by reconstitution of non‐fat dried milk powder. In all cases, oil uptake on frying with groundnut oil was not appreciably different from that of the control up to the 5% level of substitution but, at 10%, the oil uptake was higher than the control. Paneers with buttermilk and soy milk had softer textures than the control.
A review of the US Grade A raw milk supply, its quality, marketing and pricing
Authors: CHAMBERS JAMES V
Published: August 2007

As with any business, profitability is essential for the successful operation of a dairy farm. Profitability depends on the production performance of the dairy herd and a favourable price for the milk produced. While the Grade A regulations set the minimum quality standards and provide for monitoring and enforcement, the federal order system insures the producer a fair market price. Through the integration of performance standards and marketing, the United States has benefited from an abundant supply of safe and wholesome milk. However, the marketing and pricing of the US raw milk supply is beginning to model itself after the UK Milk Board concept because of mergers of cooperatives1 3 into ever larger units, increase in size of production units at the farm level and regulation of the surplus Grade A raw milk supply.
An overview of the educational needs for the international dairy industry
Authors: HENDERSON M W , TAMIME A Y
Published: August 2007

Educational provision for potential recruits to the dairy industry has changed dramatically over recent years. This study considers the current situation in various countries including the United Kingdom, and highlights the evolution of the traditional dairy based courses to curricula which now embrace science and technology of all main food areas, together with an increasingly important element of quality management. Effective delivery of current courses via the use of contemporary teaching methodologies is explored. Emerging concerns that the specific training needs of the dairy industry are poorly addressed by the nature of current courses are discussed.
Whey crystalline lactose as a main constituent in growth and counting media for lactococci and Streptococcus thermophilus
Authors: TEMIZ A , YILMAZ R
Published: August 2007

In the present study, a dried whey crystalline lactose (WCL) preparation was obtained from deproteinized whey by crystallization in an ethanol–water system. This was used as a major component in Streptococcus WCL (SWCL) broth and in Modified M17 (MM17) agar media. SWCL broth and MM17 agar were used as media for propagating and counting several strains of lactococci and Streptococcus thermophilus, respectively. The study was carried out to compare the alternative utilization of SWCL broth to RSM (reconstituted skim milk) and MM17 agar to original M17 agar. The streptococcal cultures tested in this study remained viable through several transfers in SWCL broth medium. However, high acidity development was not observed in SWCL broth cultures. The presence of 4 g/l yeast extract in SWCL broth had more effect on growth than on acid production. Growth in SWCL broth or RSM was dependent upon the type of streptococcal culture used. MM17 agar gave comparable results to M17 agar in terms of counts of the lactococci and Strep thermophilus propagated in SWCL broth or in RSM.
Concentration of acidic whey and its functionality in French type bread
Authors: YOUSIF A K , ABOU‐EISHEH M A , HUMAID M A , AL‐TABBA'A M J
Published: August 2007

Two techniques, ie, the climbing film, natural circulation evaporator and the double jacketed open pan evaporator, were used to concentrate the acidic whey. The unconcentrated (raw) whey, as well as the concentrated whey, were characterized physicochemically and incorporated into a French type (Hamam) bread dough formula as water replacers to provide 1% whey solids. The stability and the mechanical tolerance index of the French type bread dough were improved by the addition of whey solids, as were both the external and internal characteristics of the bread. The addition of whey solids also had a retardation effect on bread staling, and extended its storability by two days.
Halloumi cheese: the product and its characteristics
Authors: PAPADEMAS PHOTIS , ROBINSON RICHARD K
Published: August 2007

Halloumi, the traditional cheese of Cyprus, is extremely popular in the Middle East and the Mediterranean regions of Europe and in recent years exports from Cyprus have risen. The basic cheesemaking process places halloumi in the family of white brined cheeses, but some of its characteristics are quite unique. This paper reviews the studies that have been carried out on the chemical composition, manufacturing procedure(s), sensory quality and storage of halloumi, and considers some proposed developments, such as the use of reconstituted milk powders or homogenized milk.
Quantitative techniques in the measurement of milk adulteration in Peshawar, Pakistan**
Authors: KHAN MUNIR , RAJAH KANES K. , HAINES MICHAEL
Published: August 2007

This paper discusses the compositional parameters (fat, solids‐not‐fat and total solids) of buffalo milk sold in Peshawar, Pakistan. A total of 51 samples in phase 1 and 31 samples in phase 2 were analysed. In separate experiments, milk samples from summer and winter milkings were obtained from selected producers, and representative samples were also retrieved from further down their marketing chain for analysis. The results show that, in compositional terms, none of the market samples reached the level accepted as standard for pure buffalo milk. Therefore, it is concluded that all samples analysed had been diluted by the addition of water. The results also show that all participants in the milk marketing chain dilute milk to some extent.
Growth of thermophilic spore forming bacilli in milk during the manufacture of low heat powders
Authors: MURPHY PATRICK M , LYNCH DAVID , KELLY PHILIP M
Published: August 2007

The survival and growth of Bacillus stearothermophilus and Bacillus licheniformis, naturally present (30–300 colony forming units/ml) in late season skim milk, was monitored in a three effect evaporator during low heat skim milk powder manufacture. Substantial growth was shown to occur in the preheating stages prior to direct steam heating. A typical heat treatment (77°C, 15 s) used in the manufacture of low heat powder did not inactivate the bacteria, which continued to grow in the heater. The importance of preheaters in influencing thermophile growth in the evaporator is demonstrated by the finding that growth in the preheater stages was accompanied by growth in subsequent evaporator effects which significantly exceeded that observed when the final two preheaters were bypassed. A mid‐run mini‐clean procedure incorporating 0.2% hydrogen peroxide for decontaminating the evaporator was tested and may prove useful in extending evaporator run times
The thermostability of spray dried imitation coffee whiteners
Authors: KELLY P M , OLDFIELD D J , O'KENNEDY B T
Published: August 2007

Imitation creamer formulations were spray dried and agglomerated on a pilot scale tall‐form drier in order to evaluate the stability of the resulting powders when added to hot aqueous coffee solutions. The study explored the effects of different protein ingredients (sodium caseinate; milk protein concentrate; whey protein concentrate; milk proteinate; soluble wheat protein) in combination with non‐protein emulsifiers and disodium hydrogen orthophosphate. Adaptation of coffee stability test methodology was necessary to take account of the presence of significantly more ‘floaters’ in the case of imitation coffee whiteners which did not sediment during centrifugation. A new non‐dairy protein, soluble wheat protein, proved to have exceptional stabilizing ability compared to all other protein ingredients evaluated. Sodium caseinate performed the best out of the dairy proteins, while formulations incorporating milk protein concentrate tended to be the least stable. When working with whey protein concentrate as the principal ingredient source, an emulsifier system based on mono/diglycerides was inadequate, and it was necessary to use a combination of polysorbate and sodium stearoyl lactylate in its place.
Maribo cheese manufactured with concentrated milk: characteristics, maturation and yield
Authors: BRITO C , ASTETE M A , PINTO M , MOLINA L. H
Published: August 2007

Research was carried out to study the feasibility of making maribo cheese using milk fortified by the addition of skim milk powder. A control (T‐C) with 82 g l‐1 solids‐non‐fat (SNF) and 32 g l‐1 milk fat was included, along with three treatments with 11.7 (T‐1), 14.6 (T‐2) and 16.6 g l‐1 SNF (T‐3) and standardization of the milk fat. Some chemical characteristics of the cheese milks and of the endproducts were studied and, in addition, cheese yield and the progress of maturation were monitored. It was observed that, as maturation proceeded in all treatments, there was a steady increase in the ripening index (soluble nitrogen/total nitrogen %), which indicates a progressive advance of proteolysis. Nevertheless, there were significant differences (p <.05) between the ripening indices of the control and the rest of the treatments. Furthermore, as the extent of maturation increased, αsl‐casein was degraded more than β‐casein. The yield of cheese increased proportionally as the concentration of non‐fat‐solids in the milk increased.
Water charges: the Mogden formula explained?
Authors: WALKER STEPHEN
Published: August 2007

A series of short articles on topics of interest to the dairy industry. The subjects covered will include chemistry, consumer affairs, marketing, microbiology, nutrition and processing, and contributions of 1000–1500 words (including tables and figures) are invited from any member of the Society.
Comparison of a laboratory butter reworker with a Westfalia industrial reworker
Authors: HUERTA‐GONZALEZ LUIS , WILEEY RANDREW
Published: August 2007

Bulk butter was subjected to reworking by an industrial and a small‐scale reworker. Samples were examined by visco‐elastic rheometry in oscillatory mode. The shift in storage modulus on reworking was estimated and compared to the total applied shear (TAS) in the reworking operations. The relationship between TAS and rheological changes was similar for the two reworking systems, providing both a confirmation of the applicability of the laboratory reworker for examining butter reworking and the potential for modification of butters in the reworking process.
An approach to the detection of synthetic milk in dairy milk: 1. Detection of urea
Authors: PARADKAR MANISH M , SINGHAL REKHA S , KULKARNI PUSHPA R
Published: August 2007

A qualitative test for the detection of externally added urea (as synthetic milk) was developed using soybean urease. The quantitative detection of synthetic milk in dairy milk could be achieved by this method based on the ‘a’ value of ‘L, a, b’ scale of the HunterLab colorimeter and also by the diacetyl complexation method. Both methods are sensitive enough to detect amounts as low as 2% synthetic milk added to dairy milk, corresponding to 20 mg of added urea per 100 ml.

This work reports on methods to detect and estimate unionic detergents, which are important ingredients of synthetic milk formulations, based on the extraction of a detergent—Methylcne Blue complex in chloroform which can be subsequently quantified by measuring the optical density at 653 nm. A base value of 0.52 for optical density was found to represent pure cow's milk Any result above this value suggested the presence of an anionic detergeni in milk.
Variation in some heavy metals during the production of white cheese
Authors: DEMIRÖZÜ‐ERDINÇ B , SALDAMLI I
Published: August 2007

The lead, cadmium, arsenic and mercury contents in samples of milk, curd and white cheese taken from a production line were investigated. The lead, cadmium and arsenic levels were found to be 12.07±0.06 ng/ml, 1.82±0.01 ng/ml and 0.64±0.03 ng/ml in milk, 219.85±0.46 ng/g, 19.58±0.34 ng/g and 8.80±0.41 ng/g in cheese curd, 250.63±0.53 ng/g, 21.19±0.16 ng/g and 10.85±0.34 ng/g in fresh white cheese preripened in brine salted to 16%, and 260.25±0.51 ng/g, 22.87±0.17 ng/g and 11.35±0.29 ng/g in ripened white cheese, respectively. No detectable levels of mercury were found in any of the samples. Variation in the heavy metal contents of samples taken during the production stages was significant (p <.05). The heavy metal concentrations of the samples were compared with those obtained by other investigators in different countries.
Comparison of the characteristics of halloumi cheese made from ovine milk, caprine milk or mixtures of these milks
Authors: KAMINARIDES S , ROGOTI E , MALLATOU H
Published: August 2007

The aim of this work was to study the rheological, physicochemical and organoleptic characteristics of four types of halloumi cheese made from pure ovine milk, pure caprine milk or mixtures containing 15 or 30% caprine milk. All the milks were standardized for casein to fat ratio. Pure ovine milk gave the highest yield of halloumi cheese and caprine the lowest. No significant differences in acidity, pH, moisture, total protein and ash contents of the four types of cheese were found. In contrast, significant differences in soluble proteins, non‐protein nitrogen, acid degree value and calcium contents were noted. Rheological examinations showed the hardness and the force at the point of fracture of caprine halloumi cheese to be lower than those of the other types of cheese. The same cheese presented the highest compression at the point of fracture and cohesiveness, while there were no differences in gumminess or chewiness. Members of the assessment panel showed a preference for halloumi cheese made from 30% caprine milk and 70% ovine milk.
Manufacture of direct acidified cheese from ultrafiltration and reverse osmosis retentates
Authors: HYDAMAKA A W , WILBEY R A , LEWIS M J
Published: August 2007

Whole milk and retentates from ultrafiltration at 4:1 volume concentration ratio and reverse osmosis at 2.5:1 were used in the manufacture of direct acidified cheese. Yield based on component recovery was higher in cheese from milk retentates than whole milk. On a dry mass basis, an increase in cheese yield of 37.9% for reverse osmosis, and 14.7% for ultrafiltration was achieved compared with cheese from whole milk. Compositional variation in the resulting cheese affected both textural and sensory parameters. Cheese from ultrafiltration scored highest in sensory evaluation, although all cheeses were graded fair to good.
Reliability of findings around healthy implants in association with oral hygiene measures: a clinical, microbiological, and immunological follow‐up in edentulous patients
Authors: Lachmann Stefan , Kimmerle‐Müller Evi , Axmann Detlef , Gomez‐Roman German , Weber Heiner , Haas Robert
Published: August 2007

Objectives: To assess the performance of clinical, microbiological, and immunological diagnosis of peri‐implant health and the influence of professional hygiene measures on them.

Material and methods: Twenty‐one edentulous patients with oral implants supporting a lower overdenture were followed up over 3 months beginning 1 week before their annual recall visit. Hygiene scores, probing depth, bleeding on probing (BOP), implant stability, gingival crevicular fluid (GCF) volume, sulcular interleukin‐1β (IL‐1β) and prostaglandin E2 (PGE2) concentrations, and relative concentrations of five bacterial species (polymerase chain reaction) were investigated. Measurement variation was assessed as a function of (a) intra‐ and (b) inter‐examiner reliability, (c) inter‐implant variation in each patient, (d) time, and (e) effect of hygiene measures by accuracy, repeatability, reproducibility, and visualization with the Bland and Altman Plot.

Results: Measurement means and accuracy (in parentheses) were as follows: GCF volume 1.5 μl (1.5), Interleukin‐1β 8 ng/ml (26), PGE2 63 ng/ml (185), bacteria sum score 0.2 (0.7), plaque score 1 (1), BOP score 0 (1), Periotest value −4 (3), resonance frequency analysis ISQ 66 (11), and pocket probing depth 2.3 mm (0.7). No finding exhibited any statistically significant measurement variation as explained by accuracy, repeatability, or reproducibility. Bland and Altman Plots revealed insufficient agreement for replicated BOP assessments. A short post‐treatment reduction in plaque and BOP scores was visually apparent. Still, professional oral hygiene measures exerted no sustained influence on the clinical and biochemical appearance of the peri‐implant tissues.

Conclusion: All findings except BOP showed statistically acceptable repeatability and moderate vulnerability to influences present ‘chairside’ in clinical practice.
Body dysmorphic disorder in patients with borderline personality disorder: prevalence, clinical characteristics, and role of childhood trauma
Authors: Semiz Umit , Basoglu Cengiz , Cetin Mesut , Ebrinc Servet , Uzun Ozcan , Ergun Berk
Published: August 2007

Objective:

The prevalence of body dysmorphic disorder (BDD) in patients with borderline personality disorder (BPD) is unidentified. We hypothesised that BDD would be more common than realised in patients with BPD and comorbidity with BDD would result in a more severe clinical profile. Also, childhood trauma may play a predictive role in this association.

Methods:

The study included 70 BPD in-patients and 70 matched healthy controls. The subjects were evaluated with a comprehensive assessment battery using Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I), Structured Clinical Interview for DSM-III-R Personality Disorders (SCID-II) diagnostic tool for BDD, Global Assessment of Functioning, Traumatic Experiences Checklist and Hamilton Depression Rating Scale.

Results:

The prevalence of BDD was 54.3% in the borderline sample. The BPD patients with BDD had significantly lower overall functioning and higher levels of BPD pathology, childhood traumatic experiences, suicide attempts, substance abuse and self-mutilation than those without BDD. Traumatic experiences were significant predictor of comorbid BDD diagnosis in BPD patients.

Conclusions:

Our results suggest a relatively high prevalence of BDD among patients with BPD, and co-occurrence of BDD and BPD remains to be clarified. The additional diagnosis of BDD in patients with BPD may cause a more severe global illness, and these two disorders may share, at least partly, a common psychopathologic mechanism.
Combination of N‐acetylcysteine and metformin improves histological steatosis and fibrosis in patients with non‐alcoholic steatohepatitis
Authors: De Oliveira Claudia Pinto Marques Souza , Stefano José Tadeu , De Siqueira Erika Rabelo Forte , Silva Leonardo Soares , De Campos Mazo Daniel Ferraz , Lima Vicência Mara Rodrigues , Furuya Carlos Kioshi , Mello Evandro Sobroza , Souza Fabrício G. , Rabello Fabíola , Santos Telma Eugênio , Nogueira Monize Aydar , Caldwell Stephen H. , Alves Venâncio Avancini Ferreira , Carrilho Flair José
Published: August 2007

Aim:  There is no proven medical therapy for the treatment of non‐alcoholic steatohepatitis (NASH). Oxidative stress and insulin resistance are the mechanisms that seem to be mostly involved in its pathogenesis. The aim of our study was to evaluate the efficacy of N‐acetylcysteine (NAC) in combination with metformin (MTF) in improving the aminotransferases and histological parameters (steatosis, inflammation, hepatocellular ballooning, and fibrosis) after 12 months of treatment.

Methods:  Twenty consecutive patients (mean age 53 ± 2 years [36–68] and body mass index [BMI] 29 [25–35]) with biopsy‐proven NASH were enrolled in the study. NAC (1.2 g/day) and MTF (850–1000 mg/day) were given orally for 12 months. All patients underwent evaluation of serum aminotransferases, fasting lipid profile and serum glucose, anthropometric parameters, and nutritional status at 0 and 12 months. A low calorie diet was prescribed for all patients.

Results:  Serum alanine aminotransferase, high‐density lipoprotein, insulin, and glucose concentrations and thehomeostasis model assessment–insulin resistance (HOMA‐IR) index were reduced significantly at the end of study (P < 0.05). The BMI declined, but without statistical significance. Aspartate aminotransferase, gamma‐glutamyl transferase, alkaline phosphatase, cholesterol, and triglycerides levels were not altered with the treatment. Liver steatosis and fibrosis decreased (P < 0.05), but no improvement was noted in lobular inflammation or hepatocellular ballooning. The NASH activity score was significantly improved after treatment.

Conclusion:  Based on the biochemical and histological evidence in this pilot study, NAC in combination with MTF appears to ameliorate several aspects of NASH, including fibrosis. Further studies of this form of combination therapy are warranted to assess its potential efficacy.
Improved Surface Temperature Prediction for the Coming Decade from a Global Climate Model
Authors: Smith Doug M. , Cusack Stephen , Colman Andrew W. , Folland Chris K. , Harris Glen R. , Murphy James M.
Published: August 2007

Previous climate model projections of climate change accounted for external forcing from natural and anthropogenic sources but did not attempt to predict internally generated natural variability. We present a new modeling system that predicts both internal variability and externally forced changes and hence forecasts surface temperature with substantially improved skill throughout a decade, both globally and in many regions. Our system predicts that internal variability will partially offset the anthropogenic global warming signal for the next few years. However, climate will continue to warm, with at least half of the years after 2009 predicted to exceed the warmest year currently on record.
Divergence of Transcription Factor Binding Sites Across Related Yeast Species
Authors: Borneman Anthony R. , Gianoulis Tara A. , Zhang Zhengdong D. , Yu Haiyuan , Rozowsky Joel , Seringhaus Michael R. , Wang Lu Yong , Gerstein Mark , Snyder Michael
Published: August 2007

Characterization of interspecies differences in gene regulation is crucial for understanding the molecular basis of both phenotypic diversity and evolution. By means of chromatin immunoprecipitation and DNA microarray analysis, the divergence in the binding sites of the pseudohyphal regulators Ste12 and Tec1 was determined in the yeasts
Saccharomyces cerevisiae
,
S. mikatae
, and
S. bayanus
under pseudohyphal conditions. We have shown that most of these sites have diverged across these species, far exceeding the interspecies variation in orthologous genes. A group of Ste12 targets was shown to be bound only in
S. mikatae
and
S. bayanus
under pseudohyphal conditions. Many of these genes are targets of Ste12 during mating in
S. cerevisiae
, indicating that specialization between the two pathways has occurred in this species. Transcription factor binding sites have therefore diverged substantially faster than ortholog content. Thus, gene regulation resulting from transcription factor binding is likely to be a major cause of divergence between related species.
Augmented Wnt Signaling in a Mammalian Model of Accelerated Aging
Authors: Liu Hongjun , Fergusson Maria M , Castilho Rogerio M. , Liu Jie , Cao Liu , Chen Jichun , Malide Daniela , Rovira Ilsa I. , Schimel Daniel , Kuo Calvin J. , Gutkind J. Silvio , Hwang Paul M. , Finkel Toren
Published: August 2007

The contribution of stem and progenitor cell dysfunction and depletion in normal aging remains incompletely understood. We explored this concept in the Klotho mouse model of accelerated aging. Analysis of various tissues and organs from young Klotho mice revealed a decrease in stem cell number and an increase in progenitor cell senescence. Because klotho is a secreted protein, we postulated that klotho might interact with other soluble mediators of stem cells. We found that klotho bound to various Wnt family members. In a cell culture model, the Wnt-klotho interaction resulted in the suppression of Wnt biological activity. Tissues and organs from klotho-deficient animals showed evidence of increased Wnt signaling, and ectopic expression of klotho antagonized the activity of endogenous and exogenous Wnt. Both in vitro and in vivo, continuous Wnt exposure triggered accelerated cellular senescence. Thus, klotho appears to be a secreted Wnt antagonist and Wnt proteins have an unexpected role in mammalian aging.
Harnessing Attosecond Science in the Quest for Coherent X-rays
Authors: Kapteyn Henry , Cohen Oren , Christov Ivan , Murnane Margaret
Published: August 2007

Modern laser technology has revolutionized the sensitivity and precision of spectroscopy by providing coherent light in a spectrum spanning the infrared, visible, and ultraviolet wavelength regimes. However, the generation of shorter-wavelength coherent pulses in the x-ray region has proven much more challenging. The recent emergence of high harmonic generation techniques opens the door to this possibility. Here we review the new science that is enabled by an ability to manipulate and control electrons on attosecond time scales, ranging from new tabletop sources of coherent x-rays to an ability to follow complex electron dynamics in molecules and materials. We also explore the implications of these advances for the future of molecular structural characterization schemes that currently rely so heavily on scattering from incoherent x-ray sources.
Increased Wnt Signaling During Aging Alters Muscle Stem Cell Fate and Increases Fibrosis
Authors: Brack Andrew S. , Conboy Michael J. , Roy Sudeep , Lee Mark , Kuo Calvin J. , Keller Charles , Rando Thomas A.
Published: August 2007

The regenerative potential of skeletal muscle declines with age, and this impairment is associated with an increase in tissue fibrosis. We show that muscle stem cells (satellite cells) from aged mice tend to convert from a myogenic to a fibrogenic lineage as they begin to proliferate and that this conversion is mediated by factors in the systemic environment of the old animals. We also show that this lineage conversion is associated with an activation of the canonical Wnt signaling pathway in aged myogenic progenitors and can be suppressed by Wnt inhibitors. Furthermore, components of serum from aged mice that bind to the Frizzled family of proteins, which are Wnt receptors, may account for the elevated Wnt signaling in aged cells. These results indicate that the Wnt signaling pathway may play a critical role in tissue-specific stem cell aging and an increase in tissue fibrosis with age.
International Conservation Policy Delivers Benefits for Birds in Europe
Authors: Donald Paul F. , Sanderson Fiona J. , Burfield Ian J. , Bierman Stijn M. , Gregory Richard D. , Waliczky Zoltan
Published: August 2007

Conservation of the planet's biodiversity will depend on international policy intervention, yet evidence-based assessment of the success of such intervention is lacking. Poor understanding of the effectiveness of international policy instruments exposes them to criticism or abandonment and reduces opportunities to improve them. Comparative analyses of population trends provide strong evidence for a positive impact of one such instrument, the European Union's Birds Directive, and we identify positive associations between the rate of provision of certain conservation measures through the directive and the response of bird populations. The results suggest that supranational conservation policy can bring measurable conservation benefits, although future assessments will require the setting of quantitative objectives and an increase in the availability of data from monitoring schemes.
Use of Chimeric Antibodies as Positive Controls in an Enzyme-Linked Immunosorbent Assay for Diagnosis of Scrub Typhus (Infection by
Orientia tsutsugamushi
)
Authors: Jones Martina L. , Barnard Ross T.
Published: August 2007

The use of human sera collected from individuals of known infected and noninfected status is necessary for the validation of diagnostic assays and for the determination of cutoff values. However, the routine inclusion of pooled human sera from infected individuals for use as positive controls in commercial assay kits has many disadvantages. Sufficient quantities of sera can be difficult to obtain, and there are ethical and safety issues to be considered. Additionally, each batch of control material requires standardization, as each will differ in antibody titer. We have genetically engineered chimeric immunoglobulin G (IgG), IgM, and IgA antibodies consisting of mouse-derived variable regions and human constant regions derived from peripheral blood lymphocytes. The chimeric nature of these antibodies allows the desired antigen specificity created through mouse immunization and hybridoma technology while retaining a human constant region required for recognition by the enzyme-conjugated antihuman signal antibody. We have investigated the potential use of chimeric IgG with specificity for the major surface antigen of
Orientia tsutsugamushi
as an alternative positive control for inclusion in a commercial enzyme-linked immunosorbent assay kit for the diagnosis of rickettsia scrub typhus (caused by infection with
O. tsutsugamushi
). Chimeric IgG was expressed in stably transfected CHO cells, allowing production of unlimited quantities. The purified protein was found to have a much greater specificity for the scrub typhus antigen than the serum-derived controls. The methods described could be applied to other assay kits for the detection of antibodies against infectious agents.
Salmonella enterica
Serovar Typhi Ty21a Expressing Human Papillomavirus Type 16 L1 as a Potential Live Vaccine against Cervical Cancer and Typhoid Fever
Authors: Fraillery Dominique , Baud David , Pang Susana Yuk-Ying , Schiller John , Bobst Martine , Zosso Nathalie , Ponci Françoise , Nardelli-Haefliger Denise
Published: August 2007

Human papillomavirus (HPV) vaccines based on L1 virus-like particles (VLPs) can prevent HPV-induced genital neoplasias, the precursors of cervical cancer. However, most cervical cancers occur in developing countries, where the implementation of expensive vaccines requiring multiple injections will be difficult. A live
Salmonella
-based vaccine could be a lower-cost alternative. We previously demonstrated that high HPV type 16 (HPV16)-neutralizing titers are induced after a single oral immunization of mice with attenuated
Salmonella enterica
serovar Typhimurium strains expressing a codon-optimized version of HPV16 L1 (L1S). To allow the testing of this type of vaccine in women, we constructed a new L1-expressing plasmid, kanL1S, and tested kanL1S recombinants of three
Salmonella enterica
serovar Typhi vaccine strains shown to be safe in humans, i.e., Ty21a, the actual licensed typhoid vaccine, and two highly immunogenic typhoid vaccine candidates, Ty800 and CVD908-
htrA
. In an intranasal mouse model of
Salmonella
serovar Typhi infection, Ty21a kanL1S was unique in inducing HPV16-neutralizing antibodies in serum and genital secretions, while anti-
Salmonella
responses were similar to those against the parental Ty21a vaccine. Electron microscopy examination of Ty21a kanL1S lysates showed that L1 assembled in capsomers and capsomer aggregates but not well-ordered VLPs. Comparison to the neutralizing antibody response induced by purified HPV16 L1 VLP immunizations in mice suggests that Ty21a kanL1S may be an effective prophylactic HPV vaccine. Ty21a has been widely used against typhoid fever in humans with a remarkable safety record. These finds encourage clinical testing of Ty21a kanL1S as a combined typhoid fever/cervical cancer vaccine with the potential for worldwide application.
Differential B-Cell Responses Are Induced byMycobacterium tuberculosisPE Antigens Rv1169c, Rv0978c, and Rv1818c
Authors: Narayana Yeddula , Joshi Beenu , Katoch V. M. , Mishra Kanhu Charan , Balaji Kithiganahalli N.
Published: August 2007

The multigene PE and PPE family represents about 10% of the genome ofMycobacterium tuberculosis. Here, we report that three members of the PE family, namely, Rv1169c, Rv0978c, and Rv1818c, elicit a strong, but differential, B-cell humoral response among different clinical categories of tuberculosis patients. The study population (n= 211) was comprised of different clinical groups of both adult and child patients: group 1 (n= 94) patients with pulmonary infection, group 2 (n= 30) patients with relapsed infection, group 3 (n= 31) patients with extrapulmonary infections, and clinically healthy donors (n= 56). Among the PE proteins studied, group 1 adult patient sera reacted to Rv1818c and Rv0978c, while Rv1169c elicited immunoreactivity in group 3 children. However, all three PE antigens studied as well as the 19-kDa antigen did not demonstrate humoral reactivity with sera from group 2 patients with relapsed infection. The current study shows that while responsiveness to all three PE antigens is a good marker forM. tuberculosisinfection, a strong response to Rv0978c or to Rv1818c by group 1 adult patients with pulmonary infection or largely restricted reactivity to Rv1169c antigen in child patients with extrapulmonary infections offers the possibility of differential utility in the serodiagnosis of tuberculosis.
Comparative Analyses of Phenotypic and Genotypic Methods for Detection of Enterotoxigenic
Escherichia coli
Toxins and Colonization Factors
Authors: Sjöling Å. , Wiklund G. , Savarino S. J. , Cohen D. I. , Svennerholm A.-M.
Published: August 2007

Enterotoxigenic
Escherichia coli
(ETEC) is one of the main causes of childhood diarrhea in developing countries and in travelers. However, this pathogen has often not been reported in surveys of diarrheal pathogens, due to lack of simple standardized methods to detect ETEC in many laboratories. ETEC expresses one or both of two different enterotoxin subtypes: heat-stable toxins, a heat-labile toxin (LT), and more than 22 different colonization factors (CFs) that mediate adherence to the intestinal cell wall. Here we compare established phenotypic and genotypic detection methods and newly developed PCR detection methods with respect to sensitivity, specificity, positive predictive value, and ease of performance. The methods include GM1-enzyme-linked immunosorbent assay and dot blot techniques using specific monoclonal antibodies (MAbs) for phenotypic detection of the toxins and CFs, respectively, as well as different PCR and DNA/DNA hybridization techniques, including new PCR assays, for genotypic identification of the toxin and CF genes, respectively. We found very good general agreement in results derived from genotypic and phenotypic methods. In a few strains, LT and CFs were identified genetically but not phenotypically. Based on our analyses, we recommend initial screening for ETEC in clinical samples by multiplex toxin gene PCR. Toxin-positive strains may then be analyzed by dot blot tests for detection of the CFs expressed on the bacterial surface and by PCR for determination of additional CFs for which MAbs are currently lacking as well as for strains that harbor silent CF genes.
Persistently Infected Calves as Reservoirs for Acquisition and Transovarial Transmission of
Babesia bovis
by
Rhipicephalus
(
Boophilus
)
microplus
Authors: Howell Jeanne M. , Ueti Massaro W. , Palmer Guy H. , Scoles Glen A. , Knowles Donald P.
Published: August 2007

Babesia bovis
is a deadly disease of cattle resulting in severe economic losses in the vast regions of the world where it is endemic. If reintroduced into the United States, babesiosis would cause significant mortality in the naïve cattle population. In order to address the risk to U.S. cattle, it is essential to quantify the transovarial transmission efficiency in adult female
Boophilus microplus
ticks following acquisition feeding on persistently infected cattle. This study tested the hypothesis that infection rates are the same for larval progeny derived from females fed to repletion during persistent or acute infection. Increasing parasite levels during acute infection correlated with an increasing number of females harboring kinetes detectable in hemolymph (
r
= 0.9). The percent infected larvae ranged from 0 to 20% when derived from females fed to repletion on persistently infected calves and from 4 to 6% when derived from females fed to repletion during acute parasitemia. There was no significant difference in infection rates of larval progeny, implying that the risk associated with the introduction of either persistently infected or acutely infected cattle is equal. Parasite levels ranged from 2.4 × 10
2
to 1.9 × 10
5
in 3-day-fed larvae derived from females fed to repletion on persistently infected cattle. One group of larvae failed to transmit the parasite, suggesting that a threshold level of parasites must be obtained by larval progeny via transovarial transmission in order for larvae to deliver sufficient parasites to infect a naïve host.
Identification of Less-Common
Streptococcus pneumoniae
Serotypes by a Multiplex PCR-Based Reverse Line Blot Hybridization Assay
Authors: Zhou Fei , Kong Fanrong , Tong Zhongsheng , Gilbert Gwendolyn L.
Published: August 2007

We developed a multiplex PCR-based reverse line blot (mPCR/RLB) assay to identify 50 uncommon pneumococcal serotypes. In combination with a previously described mPCR/RLB assay (3), all 90 pneumococcal serotypes can be identified individually (32 serotypes) or, because of predictable cross-reactions, to within small groups of two to five related serotypes (58 serotypes), which can be distinguished using serotype-specific antisera.
Longitudinal Molecular Epidemiological Analysis of Feline Calicivirus Infection in an Animal Shelter: a Model for Investigating Calicivirus Transmission within High-Density, High-Turnover Populations
Authors: Coyne Karen P. , Edwards David , Radford Alan D. , Cripps Peter , Jones David , Wood James L. N. , Gaskell Rosalind M. , Dawson Susan
Published: August 2007

The control of outbreaks of calicivirus infection in high-density, high-throughput populations is a challenge to both human and veterinary medicine. In such populations, the prevalence of infection is, in part, dependent on the levels of biosecurity and how this affects virus transmission. Here we show how longitudinal analysis of feline calicivirus (FCV) infection in an animal rescue shelter can be used as a model to examine the dynamics of calicivirus transmission and evolution in such environments. FCV was isolated from 33 of 116 cats sampled over a 15-month period (overall prevalence, 28%). Sequence analysis of the immunodominant variable regions of the viral capsid gene identified 16 strains circulating in the shelter, with no single strain appearing to predominate. The majority of these strains were introduced into the shelter from the community and did not appear to be transmitted within the population. However, for three of these strains, putative transmission events within the shelter were identified. The rates of evolution within hypervariable regions of the FCV capsid gene in individual cats ranged from 0.05 to 1.4% per week, with the highest rates generally being found in animals that either acquired the virus while in the shelter or were undergoing acute infection. These data suggest that despite the high prevalence and presence of multiple strains of FCV within the shelter, the spread of such pathogens may be restricted by various control measures, including good hygiene and biosecurity.
Chronological Study of Antibiotic Resistances and Their Relevant Genes in Korean Avian Pathogenic
Escherichia coli
Isolates
Authors: Kim Tae-Eun , Jeong Yong-Wun , Cho Sun-Hee , Kim Sun-Joong , Kwon Hyuk-Joon
Published: August 2007

Antibiograms and relevant genotypes of Korean avian pathogenic
Escherichia coli
(APEC) isolates (
n
= 101) recovered between 1985 and 2005 were assessed via disc diffusion test, PCR, restriction enzyme analysis, and sequencing. These isolates were highly resistant to tetracycline (84.2%), streptomycin (84.2%), enrofloxacin (71.3%), and ampicillin (67.3%), and most of the tetracycline, streptomycin, enrofloxacin, and ampicillin resistances were associated with
tetA
and/or
tetB
,
aadA
and/or
strA-strB
, mutations in
gyrA
and/or
parC
, and
TEM
, respectively. Class 1 integrons were detected in 40 isolates (39.6%), and a variety of gene cassettes conferring streptomycin (
aadA
), gentamicin (
aadB
), and trimethoprim (
dfr
) resistances were identified:
aadA1a
(27.5%),
dfrV
-
orfD
(2.5%),
aadB
-
aadA1a
(2.5%),
dfrI
-
aadA1a
(47.5%),
dfrXVII
-
aadA5
(12.5%), and
dfrXII
-
orfF
-
aadA2
(7.5%). In addition, several types of common promoters (P
ant
) of the gene cassettes (hybrid P1, weak P1, or weak P1 plus P2) and single-nucleotide polymorphisms in
aadA1a
were identified. The results of a chronological analysis demonstrated significant and continuous increases in the frequencies of resistances to several antibiotics (tetracycline, streptomycin, enrofloxacin, ampicillin, and trimethoprim-sulfamethoxazole) and of the relevant resistance genes (
tetA
,
strA-strB
, and
TEM
), mutations in
gyrA
and
parC
, and multidrug-resistant APEC strains during the period 2000 to 2005.
Antisense Transcription in the Human Cytomegalovirus Transcriptome
Authors: Zhang Guojuan , Raghavan Bindu , Kotur Mark , Cheatham Jacquelyn , Sedmak Daniel , Cook Charles , Waldman James , Trgovcich Joanne
Published: August 2007

Human cytomegalovirus (HCMV) infections are prevalent in human populations and can cause serious diseases, especially in those with compromised or immature immune systems. The HCMV genome of 230 kb is among the largest of the herpesvirus genomes. Although the entire sequence of the laboratory-adapted AD169 strain of HCMV has been available for 18 years, the precise number of viral genes is still in question. We undertook an analysis of the HCMV transcriptome as an approach to enumerate and analyze the gene products of HCMV. Transcripts of HCMV-infected fibroblasts were isolated at different times after infection and used to generate cDNA libraries representing different temporal classes of viral genes. cDNA clones harboring viral sequences were selected and subjected to sequence analysis. Of the 604 clones analyzed, 45% were derived from genomic regions predicted to be noncoding. Additionally, at least 55% of the cDNA clones in this study were completely or partially antisense to known or predicted HCMV genes. The remarkable accumulation of antisense transcripts during infection suggests that currently available genomic maps based on open-reading-frame and other in silico analyses may drastically underestimate the true complexity of viral gene products. These findings also raise the possibility that aspects of both the HCMV life cycle and genome organization are influenced by antisense transcription. Correspondingly, virus-derived noncoding and antisense transcripts may shed light on HCMV pathogenesis and may represent a new class of targets for antiviral therapies.