bluuebunny/crossref_metadata_2025 huggingface dataset:
Multi-omics insights into host-viral response and pathogenesis in Crimean-Congo hemorrhagic fever viruses for novel therapeutic target
Authors: Neogi Ujjwal , Elaldi Nazif , Appelberg Sofia , Ambikan Anoop , Kennedy Emma , Dowall Stuart , Bagci Binnur K , Gupta Soham , Rodriguez Jimmy E , Svensson-Akusjärvi Sara , Monteil Vanessa , Vegvari Akos , Benfeitas Rui , Banerjea Akhil , Weber Friedemann , Hewson Roger , Mirazimi Ali
Published: April 2022

The pathogenesis and host-viral interactions of the Crimean–Congo hemorrhagic fever orthonairovirus (CCHFV) are convoluted and not well evaluated. Application of the multi-omics system biology approaches, including biological network analysis in elucidating the complex host-viral response, interrogates the viral pathogenesis. The present study aimed to fingerprint the system-level alterations during acute CCHFV-infection and the cellular immune responses during productive CCHFV-replication in vitro. We used system-wide network-based system biology analysis of peripheral blood mononuclear cells (PBMCs) from a longitudinal cohort of CCHF patients during the acute phase of infection and after one year of recovery (convalescent phase) followed by untargeted quantitative proteomics analysis of the most permissive CCHFV-infected Huh7 and SW13 cells. In the RNAseq analysis of the PBMCs, comparing the acute and convalescent-phase, we observed system-level host’s metabolic reprogramming towards central carbon and energy metabolism (CCEM) with distinct upregulation of oxidative phosphorylation (OXPHOS) during CCHFV-infection. Upon application of network-based system biology methods, negative coordination of the biological signaling systems like FOXO/Notch axis and Akt/mTOR/HIF-1 signaling with metabolic pathways during CCHFV-infection were observed. The temporal quantitative proteomics in Huh7 showed a dynamic change in the CCEM over time and concordant with the cross-sectional proteomics in SW13 cells. By blocking the two key CCEM pathways, glycolysis and glutaminolysis, viral replication was inhibited in vitro. Activation of key interferon stimulating genes during infection suggested the role of type I and II interferon-mediated antiviral mechanisms both at the system level and during progressive replication.
MCMBP promotes the assembly of the MCM2–7 hetero-hexamer to ensure robust DNA replication in human cells
Authors: Saito Yuichiro , Santosa Venny , Ishiguro Kei-ichiro , Kanemaki Masato T
Published: April 2022

The MCM2–7 hetero-hexamer is the replicative DNA helicase that plays a central role in eukaryotic DNA replication. In proliferating cells, the expression level of the MCM2–7 hexamer is kept high, which safeguards the integrity of the genome. However, how the MCM2–7 hexamer is assembled in living cells remains unknown. Here, we revealed that the MCM-binding protein (MCMBP) plays a critical role in the assembly of this hexamer in human cells. MCMBP associates with MCM3 which is essential for maintaining the level of the MCM2–7 hexamer. Acute depletion of MCMBP demonstrated that it contributes to MCM2–7 assembly using nascent MCM3. Cells depleted of MCMBP gradually ceased to proliferate because of reduced replication licensing. Under this condition, p53-positive cells exhibited arrest in the G1 phase, whereas p53-null cells entered the S phase and lost their viability because of the accumulation of DNA damage, suggesting that MCMBP is a potential target for killing p53-deficient cancers.
Microplastic ingestion by commercial marine fish from the seawater of Northwest Peninsular Malaysia
Authors: Foo Yuen Hwei , Ratnam Sharnietha , Lim Er Vin , Abdullah Masthurah , Molenaar Vincent J. , Shau Hwai Aileen Tan , Zhang Shoufeng , Li Hongjun , Mohd Zanuri Norlaila Binti
Published: April 2022

Over the past decade, concerns over microplastic pollution in the marine ecosystem has increasingly gained more attention, but research investigating the ingestion of microplastics by marine fish in Malaysia is still regrettably lacking. This study investigated the microplastic presence, abundance, and morphological types within the guts of four species of commercial marine fish (Atule mate, Crenimugil seheli, Sardinella fimbriata and Rastrelliger brachysoma) caught in seawater off the coast of Malaysia’s Northwest Peninsular. A total of 72 individual commercial marine fish guts from four species (fish per species n = 18) were examined. Remarkably, this study found that 100% of the samples contained microplastics. A total number of 432 microplastics (size < 5 mm) from the four species were found in the excised marine fish guts. The most common type of microplastic discovered was fragment, which accounted for 49.5% of all microplastics present. The gut microplastic content differed between species. Sardinella fimbriata recorded the greatest amount of microplastic ingestion, with an average microplastic count of 6.5 (±4.3) items per individual fish. However, there were no statistically significant differences found when comparing study species and different locations. SEM-EDX analysis confirmed the presence of microplastic particles by identifying the chemical elements found in the samples. Since the four studied species of commercial marine fish are popular protein sources in Malaysians’ daily diet, this study suggests potential microplastic exposure to humans via contaminated fish consumption in Malaysia, which was previously unknown. Based on previous scientific evidence, this study also demonstrates the high probability of microplastic ingestion in marine fish in the Malaysian seawater, which could have an adverse effect on fish health as well as marine biota.
Unlocking Andean sigmodontine diversity: five new species ofChilomys(Rodentia: Cricetidae) from the montane forests of Ecuador
Authors: Brito Jorge , Tinoco Nicolás , Pinto C. Miguel , García Rubí , Koch Claudia , Fernandez Vincent , Burneo Santiago , Pardiñas Ulyses F. J.
Published: April 2022

The Andean cloud forests of Ecuador are home to several endemic mammals. Members of the Thomasomyini rodents are well represented in the Andes, withThomasomysbeing the largest genus (47 species) of the subfamily Sigmodontinae. Within this tribe, however, there are genera that have escaped a taxonomic revision, andChilomysThomas, 1897, constitutes a paradigmatic example of these “forgotten” Andean cricetids. Described more than a century ago, current knowledge of this externally unmistakable montane rodent is very limited, and doubts persist as to whether or not it is monotypic. After several years of field efforts in Ecuador, a considerable quantity of specimens ofChilomyswere collected from various localities representing both Andean chains. Based on an extensive genetic survey of the obtained material, we can demonstrate that what is currently treated asC. instansin Ecuador is a complex comprising at least five new species which are described in this paper. In addition, based on these noteworthy new evidence, we amend the generic diagnosis in detail, adding several key craniodental traits such as incisor procumbency and microdonty. These results indicate thatChilomysprobably has a hidden additional diversity in large parts of the Colombian and Peruvian territories, inviting a necessary revision of the entire genus.
Analysis of fungal dynamic changes in the natural fermentation broth of ‘Hongyang’ kiwifruit
Authors: Zhang Zhiming , Gao Yuhong , Zhao Wenjuan , Wei Zhuo , Liu Xiaozhen , Zhang Hanyao
Published: April 2022

‘Hongyang’ kiwifruit (Actinidia chinensis Planch.) is an ideal kiwifruit wine variety. At present, there is no research on the dynamic changes of yeast during the natural fermentation of kiwifruit wine. In this study, a high-throughput was employed to analyze the fungal population composition and diversity in the samples cultured in yeast extract peptone dextrose (YPD) medium and enriched in the natural fermentation process of ‘Hongyang’ kiwifruit at four time points, day one (D1T), day three (D3T), day five (D5T), and day fifteen (D15T). Five hundred and eighty-two operational taxonomic units (OTUs) were obtained from 131 genera and 178 species samples. The diversity analysis results showed that in the early natural fermentation stage, the dominant species was Aureobasidium pullulans, and as natural fermentation proceeded, the genus Pichia became the dominant species. Pichia kluyveri was an important species at the later stages of natural fermentation. An analysis of the metabolic pathways shows that P. kluyveri plays an aromatic-producing role in the natural fermentation of ‘Hongyang’ kiwifruit. These results could provide a theoretical basis for the studies of kiwifruit fungal diversity and fungal changes during fermentation. The findings could fix a major deficiency in the production of kiwifruit fruit wine, which lacks a specific flavor-producing yeast species or strain.
Impact of Body Mass Index and Height on Hypertensive Disorders in Pregnancy
Authors: Prasad Shubham , Gaikwad Harsha S , Shekhar Himanshu , Choudhary Nishi
Published: April 2022

Introduction: Hypertension is common cause of morbidity and mortality in pregnant females. Hence, prevention and management of preeclampsia is necessity. Maternal Body Mass Index (BMI) and preeclampsia are inter-related. South Asian females are prone to obesity. Aim: To find relation among maternal BMI, height and gestational hypertension or preeclampsia, and to assess the severity of hypertensive disorders during pregnancy and maternal and foetal outcomes Materials and Methods: This prospective observational cohort study was conducted in Department of Obstetrics and Gynaecology in Vardhman Mahavir Medical College and Safdarjung Hospital (tertiary care hospital), Delhi, India, from September 2013 to December 2014. The study involved pregnant women with gestational age less than 14 weeks. After registration, body weight and height of all the subjects were measured during the first prenatal visit and recorded. Blood pressure was measured at every antenatal visit. The subjects were followed-up once monthly till 32 weeks, every 15 days till 36 weeks and weekly thereafter till delivery. To compare the baseline parameters between the two groups of patients, Chi-square test was used for categorical variables. A two-sided p-value <0.05 was statistically significant. To find out the correlation between BMI and height with pregnancy Pearson correlation coefficient test was used. Results: Total 375 consecutive pregnant women, 44 were lost to follow-up, hence, 331 were followed-up. Obese group constituted 21.8%, majority (38.1%) had normal BMI. Overweight /obese women suffered hypertensive complications more than normal and underweight women (p-value=0.001). Short height (<150 cm) pregnant females were more prone to develop hypertensive complications (p-value=0.03). The BMI (as continuous variable) was positively correlated with pregnancy induced hypertension (r=0.351; p-value <0.0001). As BMI increases, the PIH severity increases (p-value <0.0001). However, height (as continuous variable) of the patients was negatively correlated with PIH severity (r=-0.170; p-value=0.002) and as the height of patient decreases, risk of PIH severity increases. Conclusion: Short stature and high BMI pregnant females are more prone to develop hypertensive disorders and preeclampsia. Preconception prior counselling regarding weight optimisation is must.
Isolation, Identification and Antifungal Susceptibility Testing of Candida Species: A Cross-sectional Study from Manipur, India
Authors: Chongtham Urvashi , Athokpam Debina Chanu , Singh Rajkumar Manojkumar
Published: April 2022

ABSTRACT Introduction: Candidiasis is an opportunistic infection which occurs due to indiscriminate and prolonged use of broadspectrum antimicrobials, corticosteroids, immunosuppressive agents, diabetes mellitus, Human Immunodeficiency Virus (HIV), chronic renal failure, haemodialysis, renal transplantation or indwelling urinary catheter. Recently, Non-albicans Candida (NAC) species have replaced Candida albicans and emerged as an important opportunistic pathogens exhibiting decreased susceptibility to commonly used antifungal agents. Early speciation of Candida isolates along with their antifungal susceptibility testing not only will restrict the empirical use of antifungal agent but also greatly influence the treatment options for the clinicians. Aim: To speciate Candida isolates from various clinical specimens and to determine their antifungal susceptibility pattern. Materials and Methods: This study was a cross-sectional study carried out in the Mycology Section, Department of Microbiology, Jawaharlal Nehru Institute of Medical Sciences (JNIMS), Imphal, Manipur, India, from September 2016-August 2018. Candida isolates were identified using standard microbiological procedures, and speciation was done following conventional and HiCrome differential media. Antifungal susceptibility testing was determined by using Clinical and Laboratory Standards Institute (CLSI) disk diffusion method. Data analysis was done using descriptive statistics and Chi-square test. Results: A total of 100 isolates were identified from different clinical specimens, which included 43 (43%) from sputum, 34 (34%) from urine, in majority. Highest age was 92 years and lowest age was one year from whom the isolates were detected and females (57%) outnumbered males (43%) patients. Predominant Candida isolates were Candida albicans (44%), Candida tropicalis (32%). Among the azoles, the most sensitive agent was voriconazole (86%) and least was ketoconazole (56%), 81% of the total isolates were found sensitive to amphotericin B. Conclusion: The present study demonstrated that NAC spp. have surpassed Candida albicans and there is an increase in the resistance of the Candida isolates to commonly used antifungal agents. Therefore, this study highlights the need for speciation of Candida isolates upto species level and to determine the antifungal susceptibility pattern to decrease the morbidity and mortality of the patients.
Thin Layer Modelling of Indirect and Mixed-Mode on-Farm Drying of Selected Vegetables
Authors: Oji Nathaniel , Okaiyeto S. A. , Unguwanrimi Y. A.
Published: April 2022

The thin layer drying characteristics of tomato and okra slices dried using mixed-mode on-farm solar dryer, indirect mode on-farm solar dryer and open sun drying. The tomato and okra slices dried faster when dried under the mixed-mode on-farm solar dryer. Drying time was reduced considerably using the on-farm solar dryers. The drying data were fitted into Lewis, Henderson and Pabis, and page equations. The Page model (R2=0.9365, 0.9623; X2= 0.0067, 0.0000579; RMSE= 0.0086, 0.0020 and MBE= -0.008, -0.002) gave the best prediction for the mixed-mode drying and indirect mode drying of tomato slices respectively. In the same vein Page model (R2=0.9202, 0.933o; X2= 0.00091, 0.000730; RMSE= 0.0265, 0.0244 and MBE= -0.0088, -0.0074) gave the best prediction for the mixed-mode drying and indirect mode drying of okra slices respectively. Effective moisture diffusivity of tomato slices varied between -7.4724 X 10-08 and -1.6439 X 10-07 while that of okra varied between -3.12 X 10-07 and -8.08 X 10-07. The indirect mode dryer gave the best quality of dried tomato and okra.
Do Attractions Attract Tourists to a Destination? Explaining Theory with Empirical Data Based on Three Source Markets and Three Destinations
Authors: Shahzalal Md
Published: April 2022

This investigation aims to understand tourists' motivation/attraction theories based on empirical data. It analyses tourists' basic profile, their key travel facts, needs and drivers/motivations, perceptions, top activities participated at the destinations, and places visited, along with destinations' promotional activities, which may trigger visitation/repeat visitation to destinations from source markets. In the current context, Britain, New Zealand, and Singapore are applied as destinations, whereas Australia, China, and India are source markets to analyse the empirical data made publicly available by Singapore Tourism Board, VisitBritain, and Tourism New Zealand. The empirical data supports that tourists' motivations and attractions are in line with Pearce's [1] model, and attraction and marketing appeals are in line with McKercher [2] and Leiper's [3] model. Singapore is specific needs satisfier for Indian and Chinese tourists, but it is a general need satisfier for substitutable attractions for Australians. Australians are attracted to particular attractions in Britain and New Zealand. Still, Indians travel with general needs, whereas appealing to Chinese visitors by Britain and New Zealand with specific and broad attractions do work. For New Zealand, to some extent, attractions become secondary. This paper provides implications for both inbound and outbound tourism stakeholders.
SIDS, Territorial Loss, and Notions of Statehood under International Law: Reparative Approaches
Authors: Kleiman Abigail
Published: April 2022

As global climate change intensifies, Small Island Developing States are experiencing dangerous effects and facing unprecedented risk in terms of projected territorial loss. Because the international system views states as territorial entities and links state membership with the assurance of human rights, the implications of total territorial disappearance of these islands for the realization of human rights are potentially disastrous. By utilizing the metric of adherence to key principles of differentiated responsibility, the paper analyzes potential pathways for protecting vulnerable states and their inhabitants in situations of territorial loss. It concludes that any protective framework must include a reparative approach that places greater differentiated responsibility on the states that caused climate change in terms of handling migration, promoting climate resilience and adaptation, and mitigating additional climate harms, even if doing so requires significant internal change.
Forest Governance Effectiveness of Community- and Government-managed Forests in Cameroon
Authors: Basil Ewane Ewane
Published: April 2022

Effective implementation of forest governance principles such as transparency, participation, accountability, coordination, capacity, user rights protection and security driven by positive government interventions are instrumental for sustainable forest management. The study used satellite-derived data to explore forest loss in community forest (CF) and government-managed forest in Cameroon from 2001 to 2014. The study also examined the extent to which community forest rights are legally recognised and protected and how government interventions undermine or promote CF management effectiveness in Cameroon. The rationale was to understand the forest outcome of different forest management approaches with varied user rights and management objectives. This is considering that community forest managers have been seen as better managers of forests than government agencies, if their full rights are well recognised and protected. We used semi-structured open-ended questionnaires, followed by focus group interviews with CF management institutions and document review for data collection. Forest loss showed significant increasing trends and was higher in CF than in other forest management types from 2001 to 2014. Stakeholder perceptions of CF rights and government intervention or actions varied between CF management institutions. Community forest rights such as rights to full ownership and control, full management, unlimited duration, alienation, and due process and compensation are not legally recognised and/or protected in community forestry in Cameroon, producing negative forest outcomes. Negative government interventions such as imposing excessive bureaucratic obstacles and initial financial investment in the processing of application files for CF applications and commercial user rights of timber harvesting, colluding with local elites, individuals and corrupt municipal and administrative authorities, and retaining full administrative ownership and control of CF land and resources, also produces negative forest outcomes. The non- recognition and protection of higher level CF rights and excessive negative government interventions in CF management contributes to undermining community’s willingness and readiness to prevent deforestation activities and voluntary engagement in conservation activities involving tree planting and management for long term community benefits. Therefore, recognising, strengthening and protecting all the CF rights proposed by the Rights and Resources Initiative and avoiding negative government interventions in CF management are helpful strategies in achieving the expected conservation and local development objectives of CF in Cameroon and in the Congo basin region.
Comparison of Biophysical Parameters in D-DSR and TPR
Authors: Bhargav Reddy M. , Neelima T. L. , Srikanth P. , Uma Devi M. , S. Jhonson Raju S.
Published: April 2022

The study explained comparison of the biophysical parameters with in the two rice cultures. Study was conducted in the farmers field at Jogulamba Gadwal District of Telangana under same management practices. The biophysical parameters were recorded during the crop growth stages like Vegetative, Maximum Tillering, Panicle initiation, Flowering and Maturity. The data was analyzed by using the two-sample t test with equal variances reveled that there were no significant differences was observed in the dry matter produced, SPAD and LAI values of the two rice cultures at Vegetative, Maximum Tillering, Panicle initiation, Flowering and Maturity of the crop growth. The significant difference was found in the plant height at Maximum Tillering, Panicle initiation and Flowering. The total number of tillers were also significant at flowering Due to the pest infestation in the TPR reported the highest number of tillers when compared with D-DSR, the number of tillers at harvest were also less due to unproductive tillers damaged by pest in TPR than D-DSR.
A Review on Genetic Analysis of Rice (Oryza sativa L.) Crop for Yield and its Contributing Characters
Authors: Sandeep . , Dwivedi Abhishek , Kumar Sajan , Singh R. K. , Vimal S. C. , Singh Vinod , Singh Piyusha , Debnath Ashim
Published: April 2022

Rice provides major source of calories in the global population. In any crop breeding programme, presence of genetic variability is essential for broadening the gene pool. For selecting a genotype having high desirable traits, high magnitude of variability should be present in the population. The heritability estimates along with amount of genetic variability give us an idea about genetic gain due to selection. Different direct traits (number of panicle/plant, filled grains/plant and test weight) and indirect traits (tiller number/plant, plant height) determine the yield component of rice. Present review draw a clear picture of how variability influences the selection process and also how yield is associated with different direct and indirect traits. Genotypes shows high variability and desirable characters can be used in further breeding programme. A glimpse of molecular advances in rice breeding also has been incorporated in the present review.
Effect of Spacing and Plant Growth Regulators on Growth and Yield of Finger Millet (Eleusine coracana L.)
Authors: Amulya Manasa Naripogu , Umesha C.
Published: April 2022

A field experiment was carried out during Kharif season of 2021 at CRF (Crop Research Farm) of SHUATS, Prayagraj to study about the Effect of Spacing and Plant growth regulators on growth and yield Of Finger millet (Eleusine coracana L.) The test turned into specified in randomized block design by keeping three spacing levels, i.e. S - (20 x 10 cm), S2 – (30x10cm) and S3 – (40x10cm) and Plant growth regulators i.e. Boric acid 3000ppm, Gibberellic acid 50ppm and Salicylic acid 40ppm which was replicated three times. Results revealed that application of spacing 40 x 10 cm + Gibberellic acid 50ppm was recorded  significantly maximum plant height (81.62 cm), number of tillers per plant (6.16),  Plant dry weight (15.1 g/plant) and Grain yield (4.23t/ha). However,  net return (72,650 INR/ha) and B:C ratio (2.19 ) have been recorded with treatment of  40 x 10cm spacing  + Gibberellic acid 50ppm. Therefore I concluded that spacing of 40 x 10cm + Gibberellic acid 50ppm was produced more grains and economic effective.
An Analysis of 644 Upper GI Endoscopies Performed in a Nigerian Tertiary Health Institution
Authors: Israel Omolabake Bamidele , Wuam Simon
Published: April 2022

Upper GI endoscopy is an important tool in the diagnosis and management of gastrointestinal disorders and is being employed for a similar purpose in BSUTH, Makurdi. We sought to audit the practice in this institution over a 7-year period by reviewing the records in the procedural register. We found that most of the indications were diagnostic, with a few therapeutic applications. The diagnostic yield included a spectrum of benign and malignant diseases with Gastroesophageal reflux disease (GERD), gastro-duodenitis and peptic ulcer disease being the most frequent diagnosis. The therapeutic application included percutaneous gastrostomy tube placement, haemostasis of bleeding peptic ulcers, endoscopic variceal band ligation and sclerotherapy of varices, and gastro-oesophageal foreign body removal. We conclude that routine upper gastrointestinal endoscopy is feasible in our settings though, an overwhelming majority of these are diagnostic as opposed to therapeutic. There is need to expand the scope of endoscopy in our setting. Major challenges need to be addressed to sustain and expand the practice of UGI endoscopy in our setting.
Direction Finding and Tracking using Single-Ring Circular Array Antenna and Space Division Table
Authors: Park Hyeongyu , Woo Daewoong , Kim Jaesik , Park Jinsung
Published: April 2022

Single-ring circular array antennas can be applied to direction finding systems in order to use nose-section in other purposes, and the interferometry is a proper direction finding method to those systems. We usually make the interferometer baseline long enough to achieve good angular accuracy. However, an interferometer with baseline longer than a half-wavelength has the ambiguity problem. In this paper, we present a novel method for solving the ambiguity problem in interferometry systems. This technique is based on the amplitude comparison method and the space division table, and it can place a target within the angular region in which the ambiguity problem does not occur by roughly estimating direction-of-arrival. The Monte Carlo simulation results show that proposed method can effectively remove the ambiguity problem in the system.
Analysis for the Driving Dynamic Characteristics of Large Scale Semi-Trailer Equipped with Swivel Axle and Hydropneumatic Suspension Unit
Authors: Ha Taewan , Park Jungsoo
Published: April 2022

Driving dynamic characteristics of semi-trailer loaded with precise equipments are very important to protect them from vibration, impact or other disturbances. In this paper, in order to identify the driving dynamic characteristics of the large scale semi-trailer equipped with swivel axle and hydropneumatic suspension unit, Dynamics Modeling & Simulation(M&S) were performed using general Dynamics Analysis Program(RecurDyn V9R2). The semi-trailer was modeled as two types - one is Multi Rigid Body Dynamics(MRBD) model, and the other Rigid-Flexible Body Dynamics(RFlex) one. The natural vibration mode and frequencies of semi-trailer body, acceleration of dummy-weight, pitch, roll and yaw of dummy-weight, swivel axle and hydropneumatic suspension cylinder support structure, and acting force of hydropneumatic suspensions etc. were obtained from the M&S. Additionally frequency analysis were performed using the data of behavior obtained from above M&S. Generally the quantitative results of RFlex are larger than them of MRBD in view of magnitude of the comparable parametric values.
Expression of cytochrome P‐450d by Saccharomyces cerevisiae
Authors: Shimizu Toru , Sogawa Kazuhiro , Fujii-Kuriyama Yoshiaki , Takahashi Masae , Ogoma Yoshiro , Hatano Masahiro
Published: July 2002

Rat liver microsomal cytochrome P‐450d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast‐Escherichia coli shuttle vector consisting of rat liver P‐450d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P‐450d was immunologically crossed with rat liver P‐450d. The hydroxylase activity of estra‐1,3,5(10)‐triene‐3,17β‐diol was 11 nmol/min per nmol P‐450d, which is comparable to that reported previously for rat liver P‐450d. The expressed P‐450d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.
Effects of mutations at position 36 of tRNAGlu on missense and nonsense suppression in Escherichia coli
Authors: Gregory Steven T. , Dahlberg Albert E.
Published: July 2002

Mutations in the anticodon of tRNAGlu (UUC) were isolated or constructed and characterized for their ability to suppress cognate nonsense or missense mutations in vivo. The C36‐to‐A36 transversion mutation was isolated as an ochre and an amber suppressor, while the G36 transversion was selected as a CAG missense suppressor. tRNAGlu suppressors of an AAG missense mutation could not be isolated, and a U36 transition mutation introduced into tRNAGlu in vitro conferred no suppressor phenotype. Over‐expression of glutamyl‐tRNA synthetase did not increase the activity of the U36 mutant tRNAGlu, suggesting a defect at the level of translation rather than at the level of synthetase recognition.
p56lck plays a key role in transducing apoptotic signals in T cells
Authors: Di Somma M.Maddalena , Nuti Sandra , Telford John L. , Baldari Cosima T.
Published: July 2002

The CD4 receptor synergizes with the T‐cell antigen receptor (TCR) in helper T‐cell activation. However CD4 crosslinking in the absence of simultaneous TCR engagement leaves the cells primed to activation dependent apoptosis. To assess the role of the CD4 associated protein tyrosine kinase p56lck in CD4 priming to apoptosis we have constructed Jurkat T‐cell lines stably transfected with a constitutively active form of p56lck. These cells were constitutively primed to undergo apoptosis upon TCR crosslinking with specific antibodies. In addition the Jurkat JCaM1 line, which is defective for p56lck expression, was resistant to TCR induced apoptosis. These data indicate that p56lck is required for T‐cell apoptosis and that CD4 priming of T‐cells for antigen dependent apoptosis is due to inappropriate or partial activation of the p56lck signal transduction pathway.
Introduction of a CuA site into the blue copper protein amicyanin from Thiobacillus versutus
Authors: Dennison Christopher , Vijgenboom Erik , de Vries Simon , van der Oost John , Canters Gerard W.
Published: July 2002

The C‐terminal loop of the blue copper protein amicyanin, which contains three of the four active site ligands, has been replaced with a CuA binding loop. The purple protein produced has visible and EPR spectra identical to those of a CuA centre. Recent evidence strongly suggests that the CuA centre of cytochrome c oxidase and the A centre of nitrous oxide reductase are similar and are both binuclear. It therefore follows that the purple amicyanin mutant created here also possesses a binuclear CuA centre.
Co‐operative regulation of endocytosis by three RAB5 isoforms
Authors: Bucci Cecilia , Lütcke Anne , Steele-Mortimer Olivia , Olkkonen Vesa M. , Dupree Paul , Chiariello Mario , Bruni Carmelo B. , Simons Kai , Zerial Marino
Published: July 2002

Rab proteins are small GTPases involved in the regulation of membrane traffic. Rab5a has been shown to regulate transport in the early endocytic pathway. Here we report the isolation of cDNA clones encoding two highly related isoforms, Rab5b and Rab5c. The two proteins share with Rab5a all the structural features required for regulation of endocytosis. Rab5b and Rab5c colocalize with the both transferrin receptor and Rab5a, stimulate the homotypic fusion between early endosomes in vitro and increase the rate of endocytosis when overexpressed in vivo. These data demonstrate that three Rab5 isoforms cooperate in the regulation of endocytosis in eukaryotic cells.
The third nucleotide of the Gly coding triplet remembers the periodicity of the collagen chain
Authors: Makeev V.Ju. , Tumanyan V.G. , Esipova N.G.
Published: July 2002

Collagen is a fibrous protein with a primary structure with complex periodical features. We show using symbolic Fourier transform of the collagen cDNA sequence that basic periodical patterns appear there also. Strikingly they are present in the third position of triplets encoding Gly, which occupies each third position in the sequence of the protein, and to which selection on the protein level does not applied. Thus, the gene of collagen seems to appear due to pra‐gene multiplication.
Chromocystoscopy in the urological monitoring of patients with cancer of the cervix
Authors: Colombo M. , Vitobello D. , Milani R.
Published: July 2002

Objective: To determine whether chromocystoscopy could be used for the urological monitoring of cervical carcinoma. Methods: The absolute ejaculation time (AET, the interval of time between the intravenous injection of dye and the ejaculation of dyed urine in the bladder from either of the ureters) and the relative ejaculation time (RET, the difference of time between the two ejaculations) were retrospectively studied in relation to intravenous urography (IVU) and radionuclide renography (RR) in 141 patients. Results: A significant probability of normal IVU and RR was found when the AET was ≤7 min or the RET ≤4 min and when no interval occurred between the two ejaculations. Pathological IVU and RR were found when there was no ejaculation from one or both ureters after 12 min. Conclusions: Chromocystoscopy could be used for monitoring cervical cancer patients. An abnormal chromocystoscopy could indicate the need for further radiological examinations.
The α1/2 helical backbone of the prodomains defines the intrinsic inhibitory specificity in the cathepsin L‐like cysteine protease subfamily
Authors: Guo Ying Lan , Kurz Ursula , Schultz J.E , Lim Chin Chia , Wiederanders B , Schilling K
Published: July 2002

Proregions of papain‐like cysteine proteases are potent and often highly selective inhibitors of their parental enzymes. The molecular basis of their selectivity is poorly understood. For two closely related members of the cathepsin L‐like subfamily we established strong selectivity differences. The propeptide of cathepsin S was observed to inhibit cathepsin L with a K
i of 0.08 nM, yet cathepsin L propeptide inhibited cathepsin S only poorly. To identify the respective structural correlates we engineered chimeric propeptides and compared their inhibitory specificity with the wild‐types. Specificity resided in the N‐terminal part, strongly suggesting that the backbone of the prodomain was the underlying structure.
Thiol modification of diacylglycerol kinase: dependence upon site membrane disposition and reagent hydrophobicity
Authors: Czerski Lech , Sanders Charles R
Published: July 2002

Reaction rates were determined between disulfide reagents of varying hydrophobicity and single‐cysteine mutants of diacylglycerol kinase, an integral membrane protein. Polar reagents reacted most rapidly with surface‐exposed sites. However, a very non‐polar reagent also reacted more rapidly with exposed cysteines than with membrane sites. Moreover, this non‐polar reagent usually reacted more slowly with membrane sites than did more polar reagents. These results are consistent with the notion that disulfide exchange reactions involving buried cysteines of diacylglycerol kinase are very slow in the membrane interior, such that the competing rates of reactions which occur when normally buried cysteine sites make motional excursions to hydrated regions of the interface can be significant.
Molecular chaperone properties of serum amyloid P component
Authors: Coker Alun R. , Purvis Alan , Baker Douglas , Pepys Mark B. , Wood Steve P.
Published: July 2002

The selective binding of serum amyloid P component (SAP) to proteins in the pathological amyloid cross‐β fold suggests a possible chaperone role. Here we show that human SAP enhances the refolding yield of denatured lactate dehydrogenase and protects against enzyme inactivation during agitation of dilute solutions. These effects are independent of calcium ions and are not inhibited by compounds that block the amyloid recognition site on the B face of SAP, implicating the A face and/or the edges of the SAP pentamer. We discuss the possibility that the chaperone property of SAP, or its failure, may contribute to the pathogenesis of amyloidosis.
What can yeast tell us about N‐linked glycosylation in the Golgi apparatus?
Authors: Munro Sean
Published: July 2002

The N‐glycans found on eukaryotic glycoproteins occur in a vast range of different structures. A universal N‐glycan core is attached to proteins during synthesis in the endoplasmic reticulum, and then diversity is generated as the proteins pass through the Golgi apparatus. Many of the Golgi‐localised glycosyltransferases have now been identified in both yeast and mammalian cells, but it is still unclear how these enzymes are integrated into the Golgi and the rest of the cell so as to ensure efficient and specific processing of passing substrates. This review discusses the potential of the yeast system to address these issues.
Presidential administrations and the day‐of‐the‐week effect in stock returns
Authors: Liano Kartono , Liano Kadir , Manakyan Herman
Published: July 2002

This study examines the presence of a day‐of‐the‐week effect over different presidential administrations. The results indicate that the day‐of‐the‐week effect prevails during the Democratic and Republican administrations. However, the pattern of the day‐of‐the‐week effect differs between the two presidential administrations. Specifically, the negative returns on Monday are more pronounced during the Republican than during the Democratic administrations. Therefore, explanations for the day‐of‐the‐week effect should take into account the changing pattern of the day‐of‐the‐week effect across presidential administrations.
Scale‐Up Studies on a Defined Medium Process for Pilot Plant Production of Illicicolin by Gliocladiumroseum
Authors: Junker B. , Zhang J. , Mann Z. , Reddy J. , Greasham R.
Published: July 2002

Illicicolin was cultivated at the 600‐L pilot scale for purposes of material generation and process development. The initial medium containing oat flour was difficult operationally as a result of excessive foaming during sterilization, so a new defined medium process (with either glucose or sucrose as the carbon source), developed at the 23‐L scale, was scaled up and improved for pilot scale needs. Pilot scale media development efforts focused on exploring the highest concentration of media (1.0× to 3.0×) that could be cultivated at the pilot scale and not be limited by mixing or oxygen mass transfer. The process was scaled up successfully and peak titers improved 7.5‐fold, from about 200 mg/L in the initial complex medium to 1500 mg/L in the final defined medium.
Identification of Affinity Ligands for Protein Purification from Synthetic Chemical Combinatorial Libraries
Authors: Wang Yan , Li Tingyu
Published: July 2002

A method to screen combinatorial libraries for the development of selective ligands for protein affinity chromatographic purification is described. The method is based on the application of parallel combinatorial libraries, and it has several potential advantages. The screening procedure is simple and straightforward, and it does not require the chemical derivatization of the target proteins or even that the target protein be pure. The experiment can also be designed to select binders that are less likely to cause protein denaturation. Feasibility of this approach is demonstrated with a model study of the chromatographic purification of bovine albumin serum (BSA) and Avidin.
Targeting Systemically Administered Proteins to Bone by Bisphosphonate Conjugation
Authors: Uludag Hasan , Yang Jennifer
Published: July 2002

To develop a methodology for bone‐specific delivery of proteins, a bone‐seeking aminobisphosphonate (aminoBP) was previously conjugated to a model protein, bovine serum albumin (BSA). The conjugates were shown to exhibit a high affinity to bone in vitro and in vivo. This study was conducted to determine whether the systemic delivery of proteins to bone can be increased by aminoBP conjugation. Two model proteins used for this study were BSA and lysozyme (LYZ). For each protein, an unmodified (i.e., control) and aminoBP‐conjugated protein were 125I‐labeled and injected into rats, and the organ delivery of the proteins were determined. Intravenous (IV) injection of aminoBP‐BSA resulted in a 2.0‐ to 3.7‐fold increased delivery to bones as compared to the control protein in young rats. In osteopenic, ovariectomized rats, aminoBP conjugation enhanced the bone delivery of BSA by 2.2‐ to 7.5‐fold. A 3.7‐ to 5.6‐fold increased delivery was also observed for LYZ after IV injection in normal rats. In addition to IV route of administration, subcutaneous injection was also effective in delivering a higher amount of aminoBP‐conjugated proteins to bone. We conclude that conjugating bone‐seeking aminoBPs to proteins improved their delivery to mineralized tissues. The proposed targeting approach has the potential to improve the efficacy of recombinant proteins capable of stimulating bone formation by enhancing their localization to bones.
Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts
Authors: DELEHEDDE Maryse , LYON Malcolm , GALLAGHER John T. , RUDLAND Philip S. , FERNIG David G.
Published: July 2002

We examine the relationship between the chain length of heparin-derived oligosaccharides, fibroblast growth factor (FGF)-2 binding kinetics and the ability of the oligosaccharides to allow FGF-2-induced proliferation of chlorate-treated rat mammary fibroblasts. First, using an optical biosensor, we show that FGF-2 did not bind disaccharides, but definitively bound to tetrasaccharides. As the chain length increased from tetrasaccharide to octasaccharide, there was a substantial increase in kass (564000M−1·s−1 to 2000000M−1·s−1, respectively) and affinity (Kd 77nM to 11nM, respectively) for FGF-2. From decasaccharides and longer, the kass and affinity for FGF-2 was reduced substantially (tetradecasaccharide kass 470000M−1·s−1, Kd 30nM). In chlorate-treated, and hence sulphated, glycosaminoglycan-deficient cells, FGF-2 alone or in the presence of disaccharides did not stimulate DNA synthesis and it only elicited an early transient dual phosphorylation of p42/44 mitogen-activated protein kinase (MAPK). In the same cells FGF-2 in the presence of tetrasaccharides and longer oligosaccharides was able to restore DNA synthesis and enable the sustained dual phosphorylation of p42/44MAPK. However, the oligosaccharides from tetrasaccharides to octasaccharides were less potent in proliferation assays than deca- and longer oligosaccharides. Therefore, there was no correlation between the binding parameters and the potency of the oligosaccharides in DNA synthesis assays. These results demonstrate that tetrasaccharides are able to bind FGF-2 and enable FGF-2 to stimulate cell proliferation, which sets important boundary conditions for models of the FGF-2—heparan sulphate—FGF receptor complex.
Monte Carlo simulation of hyaluronidase reaction involving hydrolysis, transglycosylation and condensation
Authors: NAKATANI Hiroshi
Published: July 2002

The action of hyaluronidase on oligosaccharides from hyaluronan is complicated due to branched reaction paths containing hydrolysis, transglycosylation and condensation. The unit component of hyaluronan is a disaccharide, namely GlcA-(β1→3)-GlcNAc where GlcA and GlcNAc are d-glucuronic acid and d-N-acetylglucosamine respectively. Hyaluronan is the linear polymer formed by these disaccharide units, linked together with β1→4 glycosidic bonds. Bovine testicular hyaluronidase acts only at β1→4 glycosidic bonds of hyaluronan. The progress of product distribution from short oligosaccharides was simulated with the Monte Carlo method using the probabilistic model. The model consists only of a single enzyme molecule and a finite number of substrate and water molecules. The simulation is based on a simple reaction scheme and proceeds via an algorithm with minimum adjustable parameters generating random numbers and probabilities. The experimental data for bovine testicular hyaluronidase using [GlcA-(β1→3)-GlcNAc]4 as the starting substrate were quantitatively simulated with only three adjustable parameters. The simulated data for [GlcA-(β1→3)-GlcNAc]3 and [GlcA-(β1→3)-GlcNAc]5 as the starting substrates agreed semi-quantitatively with experimental data using the same parameters. The mechanism of the hyaluronidase reaction is a combination of branched probabilistic cycles. The condensation reaction is much weaker than the transglycosylation reaction but contributes to product distribution at the final stage of the reaction, preventing complete hydrolysis of the substrates.
Cytomegalovirus infection and non‐neutropenic fever after autologous stem cell transplantation: high rates of reactivation in patients with multiple myeloma and lymphoma
Authors: Fassas A. B.‐T. , Bolaños‐Meade J. , Buddharaju L. N. , Rapoport A. , Cottler‐Fox M. , Chen T. , Lovchik J. C. , Cross A. , Tricot G.
Published: July 2002

In a retrospective study, we examined the association between cytomegalovirus (CMV) infection and non‐neutropenic fever immediately following autologous peripheral blood stem cell transplantation for a variety of haematological malignancies and solid tumours. Sixty non‐neutropenic febrile episodes (41 in CMV‐seropositive and 19 in CMV‐seronegative patients) were evaluated. CMV reactivation, documented by CMV antigenaemia, was detected in 16 out of 41 (39%) seropositive patients compared with 0 out of 19 seronegative patients. In 12 of these 16 patients, CMV infection was considered the sole cause of fever. Thirteen patients had maximum antigenaemia levels > 5 cells/slide. Specific antiviral treatment led to the resolution of the fever in all, but two, patients, who developed fatal CMV pneumonia. Patients with multiple myeloma and lymphoma, possibly owing to a combination of disease‐related characteristics and prior immunosuppressive treatment, had high rates of CMV reactivation and may require more frequent diagnostic evaluation and prompt therapeutic intervention.
Selective enhancement of thrombopoietic activity of PEGylated interleukin 6 by a simple procedure using a reversible amino‐protective reagent
Authors: Tsunoda S. , Ishikawa T. , Watanabe M. , Kamada H. , Yamamoto Y. , Tsutsumi Y. , Hirano T. , Mayumi T.
Published: July 2002

We developed a novel method for the chemical modification of cytokines with synthetic polymers to increase the therapeutic efficacy of the former in vivo. A pH‐reversible amino‐protective reagent, dimethylmaleic anhydride (DMMAn), was used for modification of interleukin‐6 (IL‐6) with polyethylene glycol (PEG). The novel PEG‐conjugated IL‐6 (DmPEG‐IL‐6), which had been pretreated with DMMAn before PEGylation, showed up to a 140% increase in in vitro specific activity compared with PEG‐IL‐6 that had been synthesized by the previous method. Moreover, DmPEG‐IL‐6 caused thrombopoiesis more potently in mice than PEG‐IL‐6. The DmPEG‐IL‐6 Fr.1, having 3–4 PEG chains attached to the cytokine, showed the strongest thrombopoietic effect among the DmPEG‐IL‐6s with different molecular sizes that were tested. PEG‐IL‐6 Fr.1 had a 500‐fold higher potency in stimulating thrombopoiesis than native IL‐6 and DmPEG‐IL‐6 Fr.1 achieved a threefold higher thrombopoietic effect than PEG‐IL‐6 Fr.1. In addition, side‐effects, such as an increase in the plasma fibrinogen level, were not observed after injection of either PEG‐IL‐6s or DmPEG‐IL‐6s. These results suggest that PEGylation with DMMAn pretreatment may become a useful means for clinical cytokine delivery.