bluuebunny/crossref_metadata_2025_split huggingface dataset:
Refolding and simultaneous purification by three‐phase partitioning of recombinant proteins from inclusion bodies
Authors: Raghava Smita , Barua Bipasha , Singh Pradeep K. , Das Mili , Madan Lalima , Bhattacharyya Sanchari , Bajaj Kanika , Gopal B. , Varadarajan Raghavan , Gupta Munishwar N. 
Published: September 2008

AbstractMany recombinant eukaryotic proteins tend to form insoluble aggregates called inclusion bodies, especially when expressed in Escherichia coli. We report the first application of the technique of three‐phase partitioning (TPP) to obtain correctly refolded active proteins from solubilized inclusion bodies. TPP was used for refolding 12 different proteins overexpressed in E. coli. In each case, the protein refolded by TPP gave either higher refolding yield than the earlier reported method or succeeded where earlier efforts have failed. TPP‐refolded proteins were characterized and compared to conventionally purified proteins in terms of their spectral characteristics and/or biological activity. The methodology is scaleable and parallelizable and does not require subsequent concentration steps. This approach may serve as a useful complement to existing refolding strategies of diverse proteins from inclusion bodies.
Preventing serpin aggregation: The molecular mechanism of citrate action upon antitrypsin unfolding
Authors: Pearce Mary C. , Morton Craig J. , Feil Susanne C. , Hansen Guido , Adams Julian J. , Parker Michael W. , Bottomley Stephen P. 
Published: September 2008

AbstractThe aggregation of antitrypsin into polymers is one of the causes of neonatal hepatitis, cirrhosis, and emphysema. A similar reaction resulting in disease can occur in other human serpins, and collectively they are known as the serpinopathies. One possible therapeutic strategy involves inhibiting the conformational changes involved in antitrypsin aggregation. The citrate ion has previously been shown to prevent antitrypsin aggregation and maintain the protein in an active conformation; its mechanism of action, however, is unknown. Here we demonstrate that the citrate ion prevents the initial misfolding of the native state to a polymerogenic intermediate in a concentration‐dependent manner. Furthermore, we have solved the crystal structure of citrate bound to antitrypsin and show that a single citrate molecule binds in a pocket between the A and B β‐sheets, a region known to be important in maintaining antitrypsin stability.
The effects of macromolecular crowding on the mechanical stability of protein molecules
Authors: Yuan Jian‐Min , Chyan Chia‐Lin , Zhou Huan‐Xiang , Chung Tse‐Yu , Peng Haibo , Ping Guanghui , Yang Guoliang 
Published: September 2008

AbstractMacromolecular crowding, a common phenomenon in the cellular environments, can significantly affect the thermodynamic and kinetic properties of proteins. A single‐molecule method based on atomic force microscopy (AFM) was used to investigate the effects of macromolecular crowding on the forces required to unfold individual protein molecules. It was found that the mechanical stability of ubiquitin molecules was enhanced by macromolecular crowding from added dextran molecules. The average unfolding force increased from 210 pN in the absence of dextran to 234 pN in the presence of 300 g/L dextran at a pulling speed of 0.25 μm/sec. A theoretical model, accounting for the effects of macromolecular crowding on the native and transition states of the protein molecule by applying the scaled‐particle theory, was used to quantitatively explain the crowding‐induced increase in the unfolding force. The experimental results and interpretation presented could have wide implications for the many proteins that experience mechanical stresses and perform mechanical functions in the crowded environment of the cell.
Irritable bowel syndrome and dyspepsia among women veterans: prevalence and association with psychological distress
Authors: SAVAS L. S. , WHITE D. L. , WIEMAN M. , DACI K. , FITZGERALD S. , LADAY SMITH S. , TAN G. , GRAHAM D. P. , CULLY J. A. , EL‐SERAG H. B. 
Published: September 2008

SummaryBackground  The burden of functional GI disorders and their associations with psychological distress in women veterans is unclear.Aim  To examine 1‐year prevalence of irritable bowel syndrome (IBS) and dyspepsia symptoms and their associations with anxiety, depression and post‐traumatic stress disorder (PTSD) among women veterans receiving primary care at a Veteran Affairs Medical Center Women’s Clinic.Methods  Irritable bowel syndrome, dyspepsia and psychological distress were assessed using the validated self‐administered Bowel Disorder Questionnaire, the Beck Depression and Anxiety Inventories, as well as the Mississippi Scale for Combat‐Related Post‐Traumatic Stress Disorder Questionnaire.Results  We enrolled 248 women (84% participation rate). Ninety‐three (38%) reported IBS and 51 (21%) dyspepsia symptoms. Women with IBS and dyspepsia reported higher mean scores of anxiety (IBS: 24 vs. 12, P < 0.0005 and dyspepsia: 26 vs. 12, P < 0.0005), depression (IBS: 22 vs. 11, P = 0.0005 and dyspepsia: 23 vs. 11, P < 0.0005) and PTSD (IBS: 87 vs. 69, P < 0.001 and dyspepsia: 86 vs. 69, P < 0.0005). Age‐ and ethnicity‐adjusted logistic regression analyses showed a 3‐ to 46‐fold increase in odds of IBS and dyspepsia among women with anxiety, depression or PTSD.Conclusion  Women veterans have high prevalence of IBS and dyspepsia symptoms, both of which are highly associated with presence of depression, anxiety and PTSD.
The challenge of respiratory virus infections in hematopoietic cell transplant recipients
Authors: Boeckh Michael 
Published: September 2008

SummaryRespiratory virus infections in hematopoietic cell transplant (HCT) recipients are a major cause of morbidity and mortality. While respiratory syncytial virus (RSV), human metapneumonvirus, parainfluenzaviruses, and influenza viruses are well known for their potential to cause fatal pneumonia, information has only recently emerged regarding the significance of the newly discovered viruses, such as human coronaviruses NL63 and HKU1, and human bocavirus. Lymphopenia seems to be the most important risk factor for progression to lower respiratory tract disease. Airflow obstruction is another complication of respiratory virus infections after HCT, and data to date indicate this complication may occur following parainfluenza virus and RSV infection. Infection control procedures are key for prevention. Unfortunately, there are no randomized treatment studies, which make the interpretation of the literature on interventions difficult. This article reviews the spectrum of pathogens, epidemiology, risk factors and clinical manifestations of infection, as well as recent advances in diagnostic and clinical management.
Patterns of entry to community‐based services for families with children with developmental disabilities: implications for social work practice
Authors: Hiebert‐Murphy Diane , Trute Barry , Wright Alexandra 
Published: September 2008

ABSTRACTThe purpose of the present study was to understand, from parents' perspectives, the process of entry to community‐based services for children with developmental disabilities and to explore the relationship between patterns of entry and parent‐reported needs and priorities for service. Qualitative interviews were conducted with 43 mothers and 29 fathers of children with developmental disabilities approximately 6 months after the families entered community‐based childhood disability support services in Manitoba, Canada. Analysis of the coded interview data resulted in the identification of three process factors influential in the entry of families to community‐based support services: recognition of problems in child development, formal diagnosis of child disability and linkage between diagnostic services and the community‐based support system. Analysis of these processes identified four patterns of service entry associated with differing family needs for service: early entry, prompt entry, delayed entry and atypical entry. The results can be understood as a typology useful for social workers in the assessment of family needs and priorities at the point of entry to community‐based services.
Thiazolidinediones and antiblastics in primary human anaplastic thyroid cancer cells
Authors: Antonelli Alessandro , Ferrari Silvia Martina , Fallahi Poupak , Berti Piero , Materazzi Gabriele , Minuto Michele , Giannini Riccardo , Marchetti Ivo , Barani Lucio , Basolo Fulvio , Ferrannini Ele , Miccoli Paolo 
Published: September 2008

SummaryObjective  No study has evaluated the antiproliferative effects of thiazolidinediones and antiblastics in ‘primary cultured human anaplastic thyroid cancer cells’.Design  Primary anaplastic cells proliferation was evaluated after incubation with increasing concentrations of rosiglitazone or pioglitazone or antiblastics (bleomycin, cisplatin, gemcitabine) by a proliferation assay (WST‐1‐tetrazolium reaction) and cell counting.Measurements and results  A reduction of proliferation by thiazolidinediones at 1 h (from the start of tetrazolium reaction) [of 11% and 25%, with rosiglitazone, 10 or 20 (P = 0·0001) µM, respectively; of 7% and 17%, with pioglitazone, 10 or 20 (P = 0·0125) µM, respectively], and at 2 h [of 14% and 24%, with rosiglitazone, 10 (P = 0·0043) or 20 (P < 0·0001) µM, respectively; of 9% and 21%, with pioglitazone, 10 (P = 0·0397) or 20 (P = 0·0001) µM, respectively] was shown. No significant thiazolidinediones effect was observed in normal thyroid follicular cells. Bleomycin, cisplatin and gemcitabine significantly (P < 0·0001) inhibited (> 50%) anaplastic cells proliferation. Cell counting confirmed the above mentioned results. Inhibition of proliferation was similar in tumours with or without V600EBRAF mutation, both for thiazolidinediones and antiblastics.Conclusions  Thiazolidinediones exert an antiproliferative effect in primary cultured human anaplastic carcinoma cells in vitro, such as antiblastics.
Iodine status in pregnancy: role of dietary habits and geographical origin
Authors: Mian Caterina , Vitaliano Pantaleo , Pozza Dina , Barollo Susi , Pitton Mariangela , Callegari Giovanna , Di Gianantonio Elena , Casaro Anna , Acamulli Davide N , Busnardo Benedetto , Mantero Franco , Girelli Maria Elisa 
Published: September 2008

SummaryObjectives  A study was conducted on iodine status during pregnancy and its dependence on dietary habits, racial and geographical origin, and time since arrival in Italy.Design and methods  We enrolled 322 consecutive pregnant women: 217 Italians, 62 Eastern Europeans and 43 from Northern and Central Africa. All women completed a food frequency questionnaire on their dietary habits. The urinary iodide concentration (UIC) was determined in spot morning urine samples.Results  In the group as a whole, the median UIC was 83 µg/l; the UIC was < 50 in 33% and of 150 µg/l or more in 27%; it was significantly lower in Africans and Eastern Europeans than in Italians (medians 45 and 46 vs. 100 µg/l, respectively, P = 0·005). For the foreign women, there was a significant correlation between UIC and time since arrival in Italy (r = 0·22, P = 0·02). A significant link emerged between UIC and cow's milk intake (P = 0·0001). Iodine supplements were used by 40% of the women, and UIC were higher in those who did so than in those who did not (median 103 vs. 75 µg/l, P = 0·03), particularly if the latter did not drink milk (median 98 vs. 42 µg/l, P = 0·01). Multivariate analysis showed that milk was the only variable influencing UIC (OR 1·29, P = 0·0005).Conclusions  (i) Iodine levels are too low among pregnant women in our region, and particularly in foreign women. (ii) Cow's milk intake is their main source of iodine. (iii) Iodine supplementation is mandatory during pregnancy, particularly for women do not drink milk.
Surface–subsurface water exchange rates along alluvial river reaches control the thermal patterns in an Alpine river network
Authors: ACUÑA V. , TOCKNER K. 
Published: September 2008

Summary1. Water temperature is a key characteristic of stream ecosystems that is gaining scientific and managerial relevance as maximum temperatures in aquatic ecosystems increase worldwide.2. To assess the effect of surface–subsurface water exchange on stream water temperature patterns, four alluvial reaches in the Tagliamento River basin (NE Italy), constrained by geomorphic knickpoints at the upper and lower end, and two to four hyporheic flowpaths within each reach, were continuously studied during summer 2007 and winter 2007–08. Water temperature was continuously monitored at the upstream and downstream knickpoints of the floodplains, as well as at discrete upwelling areas within each reach. Discharge and vertical hydraulic gradient were measured along the alluvial reaches, and the residence time and chemistry of upwelling water were assessed four times during the study.3. Discharge variation along the study reaches revealed that massive hyporheic exchange occurred in all sites, ranging from 21% in reach 2–52% in reach 1. End member mixing analysis showed little influence of ground water, as almost all upwelling water was freshly infiltrated hyporheic water. Importantly, hyporheic exchange flows shaped surface temperature at the upwelling locations in all study reaches, providing potential thermal refugia for aquatic biota. At sites with highest hyporheic flow rates, net temperature change was also reflected at the floodplain scale.4. The magnitude of the thermal change along a hyporheic flowpath was not related to the flowpath length but to the estimated222Rn water age. Reduction in the diel thermal amplitude by hyporheic flows rather than net temperature change, reduced temperature extremes. Therefore, restoration activities to create thermal refugia should consider the role of hyporheic flows and enhance the exchange between surface and hyporheic waters.
Root canal morphology of mandibular permanent molars at different ages
Authors: Peiris H. R. D. , Pitakotuwage T. N. , Takahashi M. , Sasaki K. , Kanazawa E. 
Published: September 2008

AbstractAim  To investigate differences in the root canal morphology of permanent mandibular molar teeth at various ages.Methodology  Four hundred and eighty permanent mandibular first and second molars were examined. First and second molars were divided into six and five groups, respectively, according to the age of the patient at the time of extraction. Root canal morphology was studied using a clearing technique. The canal morphology of the mesial root was classified into three stages depending on its developmental pattern. When the root canal system was completely differentiated, the canal classification and the number of lateral canals and inter‐canal communications were recorded. Vertucci’s classification was taken as the main reference. Canal morphology was compared amongst age groups.Results  In both first and second molars, developmental stages of canal morphology amongst age groups were significantly different (P < 0.0001). The prevalence of inter‐canal communications was highly significantly different in the first (P < 0.0001) and less significant in the second molar (P < 0.05). After completion of the canal differentiation, the mesial roots of first molars had type IV and II canal forms. The majority of the mesial roots of second molars had type I and III canals. C‐shaped canals were found in 3% of second molars.Conclusions  Mesial roots of first and second molars mostly had one large canal until 11 and 15 years of age, respectively. In both molars, the canal system was completely defined at 30–40 years. The prevalence of inter‐canal communications was low at young and old ages but high at intermediate ages.
Substance P influenced gelatinolytic activity via reactive oxygen species in human pulp cells
Authors: Wang F.‐M. , Hu T. , Cheng R. , Tan H. , Zhou X.‐D. 
Published: September 2008

AbstractAim  To investigate the effects of substance P (SP) on gelatinolytic activity of matrix metalloproteinases (MMPs) in human pulp cells.Methodology  Human dental pulp cells were isolated and cultured. Subconfluent cells, between the third and sixth passages, were maintained under serum deprivation for 18 h followed by the treatment of varying doses of SP (1 pmol L−1, 100 pmol L−1, 10 nmol L−1, 1 μmol L−1 and 100 μmol L−1). Conditioned media were then subjected to gelatin zymography using 8% sodium dodecyl sulphate polyacrylamide gel electrophoresis minigels containing 1.5 g L−1 gelatin. The effect of SP on intracellular reactive oxygen species (ROS) was also examined by confocal microscopy. ROS scavenger N‐Acetyl‐l‐cysteine (NAC, 5 mmol L−1) was utilized to evaluate the roles of ROS pathway in mediating the impact of SP on cellular gelatinolytic activity. Data were analysed using analysis of variance with Bonferroni correction for multiple comparisons or an unpaired Student’s t‐test.Results  Substance P, at levels above 1 μmol L−1, remarkably enhanced MMP‐2 activity reflected by the band migrating at 66 kDa (P < 0.05). A gelatinolytic band at approximately 44 kDa appeared to be intensified in a SP dose‐dependent manner. In addition, it was demonstrated that SP could induce ROS production in pulp cells and ROS scavenger NAC was further found to significantly reduce MMP‐2 activity (P < 0.05), as well as other bands of gelatinolytic proteinases.Conclusion  Substance P can influence gelatinolytic activity in human pulp cells via ROS pathway.
Dentinal tubule invasion and adherence by Enterococcus faecalis
Authors: Chivatxaranukul P. , Dashper S. G. , Messer H. H. 
Published: September 2008

AbstractAim  To investigate dentinal tubule invasion and the predilection of Enterococcus faecalis for dentinal tubule walls.Methodology  The invasion of dentinal tubules in extracted human teeth by E. faecalis was measured ex vivo after 8 weeks of incubation. The canal walls of 16 root sections were either intact or instrumented with or without smear layer present. Extent and maximum depth of tubule invasion were assessed histologically and compared between groups. In the adherence study, 44 vertically split root samples were prepared to expose longitudinally aligned dentinal tubules and fractured orthodentine (OD). Surfaces were exposed to E. faecalis (erythromycin resistant strain, JH2‐2 carrying plasmid pGh9:ISS1) and incubated aerobically for 2 h. Samples were processed for analysis using scanning electron microscopy. Bacterial adhesion to tubule walls versus fractured OD was calculated as number of cells per 100 μm2.Results  The strain of E. faecalis used in this study showed moderate to heavy tubule invasion after 8 weeks. In the adhesion studies, significantly more bacteria adhered to fractured OD than to dentinal tubule walls (anova, P < 0.001). With respect to the tubule wall, adherence was greater in inner versus outer dentine (P = 0.02) and greater when bacterial adhesion was tested in chemically defined medium than in phosphate‐buffered saline (anova, P < 0.001).Conclusions  Although E. faecalis readily invaded tubules, it did not adhere preferentially to tubule walls. Initial colonization of dentinal tubules by E. faecalis may depend primarily on other factors.
Application of ISO22000 and comparison to HACCP for processing of ready to eat vegetables: Part I
Authors: Varzakas Theodoros H. , Arvanitoyannis Ioannis S. 
Published: September 2008

SummaryPreliminary Hazard Analysis was used to analyse and predict the occurring failure modes in a food chain system (ready to eat vegetables processing plant), in conjunction with ISO22000, the new Food Safety Management System, based on the functions, characteristics and/or interactions of the ingredients or the processes, upon which the system depends. Critical control points (CCPs) have been identified and implemented in the Hazard Analysis Critical Point Control plan. The decision table for CCP determination during processing of ready to eat vegetables is shown and compared with the ISO22000 Analysis Worksheet for determination of the prerequisite programmes. The prerequisite programmes are the main difference between the two systems. The incorporation of PrPs in the ISO22000 made the system more flexible as a smaller number of CCPs was introduced.
Optimisation of supercritical carbon dioxide extraction of lutein esters from marigold (Tagetes erect L.) with soybean oil as a co‐solvent
Authors: Ma Qingxiang , Xu Xiang , Gao Yanxiang , Wang Qi , Zhao Jian 
Published: September 2008

SummaryResponse surface methodology was used to optimise the conditions of supercritical carbon dioxide extraction of lutein esters from marigold (Tagetes erect L.) with soybean oil as a co‐solvent. The effect of pressure, temperature, CO2 flow rate and soybean oil concentration, and their interactions on the yield of lutein was investigated. Results showed that the data could be well fitted to a second‐order polynomial model with a R2‐value of 0.9398. The independent parameters of flow rate, pressure, temperature, all quadratics as well as the interactions between flow rate and pressure, and between flow rate and temperature affected the yield significantly (P ≤ 0.05). The model predicted that the optimal conditions were 35.5 MPa, 58.7 °C, CO2 flow rate of 19.9 L/h with 6.9% of soybean oil as a co‐solvent, and under such conditions, the maximum yield of 1039.7 mg lutein /100 g marigold could be achieved.
Effect of preliminary and culinary processing on amino acid content and protein quality in frozen French beans
Authors: Kmiecik Waldemar , Lisiewska Zofia , Słupski Jacek , Gębczyński Piotr 
Published: September 2008

SummaryThe aim of the study was to evaluate the content of amino acids and protein quality of French bean pods. The investigated material consisted of the raw vegetable, fresh pods cooked to consumption consistency, and two kinds of frozen products stored for 12 months at −20 °C and then prepared for consumption: frozen beans obtained using the traditional method (blanching before freezing) and frozen products of the ready‐to‐eat type (cooking before freezing). A comparison of the amino acid content in the product prepared for consumption showed that the lowest quantities were found in French beans obtained using the traditional method; products obtained using the modified technology and beans cooked directly after harvest had similar levels of most amino acids. The content of amino acids in 16 g N was less varied than in 100 g of the product. The protein in all the three products prepared for consumption as well as that in fresh bean pods hardly differs, as confirmed by the values in the essential amino acid index (EAA). The first limiting amino acid was methionine with cystine and the second was lysine.