bluuebunny/crossref_metadata_2025_split huggingface dataset:
The absence of molybdenum cofactor sulfuration is the primary cause of the flacca phenotype in tomato plants
Authors: Sagi Moshe , Scazzocchio Claudio , Fluhr Robert 
Published: March 2003

SummaryThe molybdenum cofactor (MoCo)‐containing enzymes aldehyde oxidase (AO; EC 1.2.3.1) and xanthine dehydrogenase (XDH; EC 1.2.1.37) require for activity a sulfuration step that inserts a terminal sulfur ligand into the MoCo. The tomato flacca mutation was originally isolated as a wilty phenotype due to a lack of abscisic acid (ABA) that is related to simultaneous loss of AO and XDH activities. An expressed sequence tag candidate from tomato was selected on the basis of homology to sulfurases from animals, fungi and the recently isolated Arabidopsis genes LOS5/ABA3. The tomato homologue maps as a single gene to the bottom of chromosome 7, consistent with the genetic location of the flacca mutation. The structure of FLACCA shows a multidomain protein with an N‐terminal NifS‐like sulfurase domain; a mammal‐specific intermediate section; and a C‐terminus containing conserved motifs. Prominent among these are molybdopterin oxidoreductases and thioredoxin redox‐active centre/iron–sulfur‐binding region signatures which may be relevant to the specific sulfuration of MoCo. Indeed, the molecular analysis of flacca identifies the mutation in a highly conserved motif located in the C‐terminus. Activity gel assays show that FLACCA is expressed throughout the plant. Transient and stable complementation of flacca and the Arabidopsis aba3 mutants with Aspergillus nidulans hxB and FLACCA yielded full, partial and tissue‐specific types of Mo‐hydroxylase activities. Restoration of activity in the root alone is sufficient to augment plant ABA content and rectify the wild‐type phenotype. Thus the pleiotropic flacca phenotype is due to the loss of activity of enzymes requiring a sulfurated MoCo.
Transgenic crops, biotechnology and ownership rights: what scientists need to know
Authors: Kowalski Stanley P. , Ebora Reynaldo V. , Kryder R. David , Potter Robert H. 
Published: March 2003

SummaryOwnership of intellectual and tangible property (IP/TP) rights in agricultural biotechnology (ag‐biotech) and transgenic plants has become critically important. For scientists in all institutions, whether industrialized or developing country, public or private sector, an understanding of IP/TP rights is fundamental in both research and development. Transgenic plants and ag‐biotech products embody numerous components and processes, each of which may have IP/TP rights attached. To identify these rights, a transgenic plant or ag‐biotech product must be dissected into its essential components and processes, with each ‘piece’ analysed under the IP/TP ‘microscope’. This product deconstruction is an integral step in product clearance (PC) analysis leading to freedom to operate (FTO). To facilitate a PC analysis, the following points are important: (1) knowing what one has and where it's from, (2) organizing material transfer agreements and licences, (3) researching scientific and patent databases and relevant literature, (4) instituting a laboratory notebook policy, (5) keeping track of ownership of germplasm and plant genetic resources, and (6) promoting ongoing IP/TP management, awareness and training. However, a FTO opinion does not solve the IP/TP issues of releasing a transgenic plant or ag‐biotech product; rather, it is a management tool for assessing the risks of litigation. When transferring transgenic plants or ag‐biotech to developing nations, scientists from industrialized countries have the heightened responsibility of verifying that IP/TP issues are fully addressed and documented. Successful technology transfer goes beyond research, development and licensing; it is an holistic package leading to long‐term partnerships in international development. Managing IP/TP requires capacity‐building in scientists and technology transfer offices, in both industrialized and developing countries.
Key amino acid residues required for aryl migration catalysed by the cytochrome P450 2‐hydroxyisoflavanone synthase
Authors: Sawada Yuji , Kinoshita Kengo , Akashi Tomoyoshi , Aoki Toshio , Ayabe Shin‐ichi 
Published: March 2003

SummaryIsoflavonoids are distributed predominantly in leguminous plants, and play pivotal roles in the interaction of host plants with biological environments. Isoflavones in the diet also have beneficial effects on human health as phytoestrogens. The isoflavonoid skeleton is constructed by the CYP93C subfamily of cytochrome P450s in plant cells. The reaction consists of hydroxylation of the flavanone molecule at C‐2 and an intramolecular 1,2‐aryl migration from C‐2 to C‐3 to yield 2‐hydroxyisoflavanone. In this study, with the aid of alignment of amino acid sequences of CYP93 family P450s and a computer‐generated putative stereo structure of the protein, candidates for key amino acid residues in CYP93C2 responsible for the unique aryl migration in 2‐hydroxyisoflavanone synthase reaction were identified. Microsomes of recombinant yeast cells expressing mutant proteins of CYP93C2 were prepared, and their catalytic activities tested. The reaction with the mutant in which Ser 310 in the centre of the I‐helix was converted to Thr yielded increased formation of 3‐hydroxyflavanone, a by‐product of the 2‐hydroxyisoflavanone synthase reaction, in addition to the major isoflavonoid product. More dramatically, the mutant in which Lys 375 in the end of β‐sheet 1–4 was replaced with Thr produced only 3‐hydroxyflavanone and did not yield the isoflavonoid any longer. The roles of these amino acid residues in the catalysis and evolution of isoflavonoid biosynthesis are discussed.
Rapid identification of Arabidopsis insertion mutants by non‐radioactive detection of T‐DNA tagged genes
Authors: Ríos Gabino , Lossow Andrea , Hertel Britta , Breuer Frank , Schaefer Sabine , Broich Melanie , Kleinow Tatjana , Jásik Ján , Winter Jochen , Ferrando Alejandro , Farrás Rosa , Panicot Mireia , Henriques Rossana , Mariaux Jean‐Baptist , Oberschall Attila , Molnár Gergely , Berendzen Kenneth , Shukla Vijaya , Lafos Marcel , Koncz Zsuzsanna , Rédei George P. , Schell Jeff , Koncz Csaba 
Published: March 2003

SummaryTo assist in the analysis of plant gene functions we have generated a new Arabidopsis insertion mutant collection of 90 000 lines that carry the T‐DNA of Agrobacterium gene fusion vector pPCV6NFHyg. Segregation analysis indicates that the average frequency of insertion sites is 1.29 per line, predicting about 116 100 independent tagged loci in the collection. The average T‐DNA copy number estimated by Southern DNA hybridization is 2.4, as over 50% of the insertion loci contain tandem T‐DNA copies. The collection is pooled in two arrays providing 40 PCR templates, each containing DNA from either 4000 or 5000 individual plants. A rapid and sensitive PCR technique using high‐quality template DNA accelerates the identification of T‐DNA tagged genes without DNA hybridization. The PCR screening is performed by agarose gel electrophoresis followed by isolation and direct sequencing of DNA fragments of amplified T‐DNA insert junctions. To estimate the mutation recovery rate, 39 700 lines have been screened for T‐DNA tags in 154 genes yielding 87 confirmed mutations in 73 target genes. Screening the whole collection with both T‐DNA border primers requires 170 PCR reactions that are expected to detect a mutation in a gene with at least twofold redundancy and an estimated probability of 77%. Using this technique, an M2 family segregating a characterized gene mutation can be identified within 4 weeks.
Silica deposition by a strongly cationic proline‐rich protein from systemically resistant cucumber plants
Authors: Kauss Heinrich , Seehaus Kai , Franke Rochus , Gilbert Sabine , Dietrich Robert A. , Kröger Nils 
Published: March 2003

SummaryInfection of one leaf of cucumber (Cucumis sativa) plants can render other leaves resistant to various pathogens. This so‐called systemic acquired resistance (SAR) can be functionally mimicked by certain chemicals. All these treatments enhanced expression of a gene encoding a novel proline‐rich protein (PRP1) which has C‐terminal repetitive sequences containing an unusually high amount of lysine and arginine residues. Antibodies raised against a synthetic peptide derived from four of the repetitive sequences cross‐reacted mainly with a cell wall polypeptide of an apparent MW of 8 kDa. The protein accumulated upon SAR induction, though it does not appear to take part in oxidative protein cross‐linking, at least in the hypocotyl tissue. The synthetic peptide derived from the repetitive sequences was able to polymerize orthosilicic acid to insoluble silica, a property not resulting directly from the primary protein sequence, but rather from the high positive charge density. Our results suggest that during induction of SAR, the synthesis of a strongly cationic PRP prepares the cell walls for enhanced silica deposition which is known to participate in cell wall reinforcement, localized at the site of attempted penetration of fungi into epidermal cells. Constitutive accumulation of related PRPs may function in silica deposition during certain developmental stages, a process important for various physiological functions of green plants.
Proliferating Floral Organs (Pfo), a Lotus japonicus gene required for specifying floral meristem determinacy and organ identity, encodes an F‐box protein
Authors: Zhang Shulu , Sandal Niels , Polowick Patricia L. , Stiller Jiri , Stougaard Jens , Fobert Pierre R. 
Published: March 2003

SummaryTo study flower development in the model legume Lotus japonicus, a population of transgenic plants containing a maize transposable element (Ac) in their genome was screened for floral mutants. One mutation named proliferating floral organs (pfo) causes plants to produce a large number of sepal‐like organs instead of normal flowers. It segregates as a single recessive Mendelian locus, and causes sterility. Scanning electron microscopy revealed that pfo affects the identity, number and arrangement of floral organs. Sepal‐like organs form in the first whorl, and secondary floral meristems are produced in the next whorl. These in turn produce sepal‐like organs in the first whorl and floral meristems in the second whorl, and the process is reiterated. Petals and stamens are absent while carpels are either absent or reduced. The pfo phenotype was correlated with the presence of an Ac insertion yielding a 1.6‐kb HindIII restriction fragment on Southern blots. Both the mutant phenotype and this Ac element are unstable. Using the transposon as a tag, the Pfo gene was isolated. Conceptual translation of Pfo predicts a protein containing an F‐box, with high overall similarity to the Antirrhinum FIMBRIATA, Arabidopsis UNUSUAL FLORAL ORGANS and Pisum sativum Stamina pistilloida proteins. This suggests that Pfo may regulate floral organ identity and meristem determinacy by targeting proteins for ubiquitination.
Clinical use of FFP: results of a retrospective process and outcome audit
Authors: Jones  , Jones  , Napier  , Al‐Ismail  
Published: March 2003

An audit has been carried out on the usage of 216 units of fresh frozen plasma (FFP) issued to 41 patients. This involved the systems of FFP issue, the appropriateness of prescription as well as the recorded benefits. Sixty‐six per cent of the initial requests for FFP appeared to satisfy criteria for appropriate use. Review of the case notes resulted in some changes to earlier decisions and a slight increase to 73% of those accepted as valid. Only 94% of the FFP issued could be proved to have been given to identified patients and only 88% of case notes showed the reason for treatment.Pre‐ and post‐treatment coagulation results were available for all patients. In 15% of cases, pretreatment results were not significantly abnormal, and consequently no post‐treatment improvement found. Coagulation improvement was documented in 78% of cases, and clinical statements on patient progress noted in 40%.In 80% of cases, the patient received other blood products carrying a potential virus risk. Six of the remaining eight patients were treated to stabilize oral anticoagulation. For these patients, virus‐inactivated prothrombin concentrates could have been used, resulting in almost the same reduction in virus transmission risk for the group of 41 patients as could be obtained by using virus‐inactivated FFP.
Unchanging attitudes to autologous transfusion in the UK
Authors: Torella F. , Haynes S. L. , Lardi A. , O'Dwyer S. T. , McCollum C. N. 
Published: March 2003

. Our aim was to assess changes in attitudes to autologous transfusion amongst surgeons over a 10‐year period in response to scientific evidence, public awareness, published guidelines, management and the increasing cost of blood products.Surgeons across the north‐west of England completed questionnaires on knowledge, experience and attitude towards autologous transfusion in 1990, 1994 and 1999.Main outcome measures were changes in knowledge, experience and utilization of autologous transfusion; perceived advantages of autologous transfusion, obstacles to its implementation in surgical practice and preferences for specific techniques (preoperative autologous donation, acute normovolaemic haemodilution, intraoperative and postoperative cell salvage).There has been little change in practice over 10 years. Many more surgeons were keen to employ autologous transfusion than were using it. Autologous transfusion was only used in general, orthopaedic and cardiothoracic surgery. Safety and patient preference were the main arguments for implementation and logistics the main obstacles.Autologous transfusion was used sporadically in surgical practice. Clinical trials are needed to guide clinicians in the choice of transfusion techniques.
Polymorphism and recombination events at theABOlocus: a major challenge for genomic ABO blood grouping strategies
Authors: Olsson M. L. , Chester M. A. 
Published: March 2003

. The blood groupABOgene codes for a glycosyltransferase that adds the ultimate monosaccharide to a glycoconjugate and forms the A or B blood group specific antigen. The DNA structure of the three major alleles of the human blood group ABO system was first described in 1990. This review describes the subsequent developments, including the increasing number of variants of these common alleles and the underlying mutations thought to be responsible for the occurrence of some of the weak subgroups of blood group A and B. Several inactive (O) alleles are also now known. Our knowledge of the DNA sequence of the normalAandBalleles and of the rare and intriguingcisAB and B(A) phenotypes has resulted in plausible explanations for these.Allelic variations outside the translated exons have been investigated and resulted in detection of lineage‐specific intron mutations and the discovery of an enhancer VNTR region affecting the rate of transcription at this locus.The occurrence of hybrid alleles can also explain hitherto abnormal inheritance in some pedigrees. The detection of hybrid alleles has been made possible by the presence of numerous polymorphisms found in the variousABOalleles. The role ofchi(χ) sequences is discussed.Finally, the various genotyping methods available are summarized and their advantages and limitations are analysed in the light of the increasing allelic variation.
Outcome of consecutive pregnancies in a patient with Bombay (Oh) blood group
Authors: Bhattacharya S. , Makar Y. , Laycock R. A. , Gooch A. , Poole J. , Hadley A. 
Published: March 2003

Summary A young lady with a rare Bombay (Oh) blood group had two successive uneventful pregnancies. Her serum contained a potent high‐titre anti‐H and serological as well as chemiluminescence tests, suggesting that the antibody was haemolytic. Her husband was of the normal H status. Theoretically, both babies should have been positive for the H antigen and should have suffered from haemolytic disease of the newborn.This apparent conundrum could be owing to the weak expression of the H antigens on the infant red cells.
Trichomoniasis treatment in women: a systematic review
Authors: Gülmezoglu A. Metin , Garner Paul 
Published: March 2003

objective To compare the effectiveness of various treatment strategies for trichomoniasis in women.data sources Medline from 1966 to1996, Embase from 1986 to 1996, Science Citation Index from 1990 to 1996; reference lists of existing reviews; through the manufacturers of metronidazole and tinidazole in the UK, the Cochrane Controlled Trials Register until October 1997 and informal discovery.study selection Any randomized or quasi‐randomized trial in nonpregnant women with trichomoniasis where different treatment strategies were compared. 45 of the 124 identified studies met the criteria and were included in the review.data extraction Settings, diagnostic methods, exclusions, loss to follow‐up and partner treatment strategies were extracted. Outcomes sought were parasitological cure, clinical cure and side‐effects of treatment.results Most trials were small, with only two trials containing more than 100 women in each comparison group. Only 11 trials followed up women for more than one month. Oral nitroimidazoles were effective in achieving parasitological cure. Fourteen trials compared different treatment strategies with the remainder comparing different doses or different drugs. Partner treatment was effective in decreasing longer‐term re‐infection rates in the one trial testing this.conclusions Parasitological cure can be achieved by a single oral dose of nitroimidazoles. There is, however, very little data on partner treatment strategies and long‐term cure rates after initial treatment. Further research should test various partner treatment strategies to prevent re‐infections and reduce trichomoniasis prevalence.
Characterization of secretory acetylcholinesterase from Setaria cervi microfilariae: a potential antigen for diagnosis of human filariasis
Authors: Sharma Sudha , Rathaur Sushma 
Published: March 2003

Summary Acetylcholinesterase (AChE) is released to the external medium when microfilariae (mf) of Setaria cervi, a bovine filarial parasite, are maintained in vitro. Intense enzyme staining at amphids, excretory pores, anal vesicle and phasmids suggest an active secretion of AChE from mf. Excretory‐secretory products of mf displayed two electromorphic variants of AChE when resolved by 6% nondenaturing PAGE. The two isoforms of AChE (A and B) were separated on the basis of charge by DEAE sepharose CL 6B column following gel filtration. The two isoforms showed differing kinetic properties with respect to substrate specificity and inhibitor sensitivity. Anti‐Nippostrongylus brasiliensis AChE antibodies cross‐reacted with the affinity purified secretory AChE in ELISA. Immunoblotting of purified AChEs with cross‐reacting anti‐AChE antibodies revealed the presence of an approximately 75 kD protein in the isoenzyme A and an approximately 45 kD protein in B, whereas both proteins were present in the enzyme purified via affinity chromatography on edrophonium sepharose column.
Parameters associated with Schistosoma haematobium infection before and after chemotherapy in school children from two villages in the Coast province of Kenya
Authors: Kahama Anthony I. , Vennervald Birgitte J. , Kombe Yeri , Kihara Ruth W. , Ndzovu Malick , Mungai Peter , Ouma John H. 
Published: March 2003

Summary We evaluated the impact of praziquantel therapy (40 mg/kg body weight) on indicators of infection with Schistosoma haematobium by following a cohort of infected children from schools located 12 km apart in the Coast province of Kenya, at 0, 2, 4, 6, 12 and 18 months after treatment. Within this period, measurements of infection parameters pertaining to egg counts and haematuria (micro‐, macro‐ and history) were evaluated at all time points. The initial prevalence of 100% dropped significantly 8 weeks after treatment with a similar trend in the intensity of infection. Microhaematuria followed the same trend as observed for egg counts while macrohaematuria remained low after treatment. Reinfection following successful therapy differed significantly between schools; in one school the children were reinfected immediately while those in the other remained uninfected despite similar starting prevalences, intensities of infection and cure rates. Transmission between the two areas looked homogeneous before treatment but when both groups were treated, contrasting transmission patterns became evident. In a regression model we evaluated factors that might be associated with reinfection, and after allowing for pretreatment infection level, age and sex, area (school) remained a highly significant predictor.
Hantaan virus antibody prevalence in rodent populations of several provinces of northeastern Thailand
Authors: Nitatpattana Narong , Henrich Timothy , Palabodeewat Somnuek , Tangkanakul Waraluk , Poonsuksombat Duangporn , Chauvancy Gilles , Barbazan Philippe , Yoksan Sutee , Gonzalez Jean‐Paul 
Published: March 2003

SummaryWe conducted a serological survey of 632 rodents from the northeast region of Thailand in order to assess the presence of Hantaan‐like viruses that may be a risk to the human population. Rodents were collected from rice fields, houses and domestic gardens in five northeastern provinces and tested for IgG reacting sera to Hantaan antigen using enzyme‐linked immunoassays. The overall prevalence of Hantavirus infection in rodents was 2.1% (13/632). Species that tested positive included Bandicota indica (4.3% positive within species), Rattus exulans (2.1%), R. losea (1.6%) and R. rattus (0.9%). Species such as R. exulans and R. losea are candidate hosts of unidentified Hantaan‐like viruses in Thailand.
Health service use and household expenditure during terminal illness due to AIDS in rural Tanzania
Authors: Ngalula Juliana , Urassa Mark , Mwaluko Gabriel , Isingo Raphael , Ties Boerma J. 
Published: March 2003

SummaryMost knowledge about health seeking behaviour during terminal illness among people with HIV/AIDS in Africa is derived from health facility based studies. This study uses data from a longitudinal community study in a rural area in north‐west Tanzania, where interviews were conducted on health seeking behaviour and expenditure with relatives who lost an adult family member in recent months. HIV status and verbal autopsy were used to assess if service use differed by cause of death. During terminal illness, people with HIV/AIDS made extensive use of both traditional and modern health services, and more so than people who died from other causes. The main factor associated with this difference was the longer duration of illness. Expenses associated with HIV/AIDS terminal illness were higher than for other causes of death, largely because of the longer duration of illness. The direct medical costs were about 1.5 times higher than the funeral costs and the sum of the medical and funeral costs exceeded the estimated annual household income per capita in this population. In conclusion, the rapid increase in numbers of terminally ill adults as a result of HIV/AIDS is likely to lead to an increased burden on all layers of the health system and household resources, in part because of the relatively long duration of HIV/AIDS terminal illness. However, almost half of all HIV/AIDS deaths in this rural population were not admitted to hospitals during their terminal illness and only a small proportion died in hospitals. If more effective treatment becomes available, a further increase in health service use and direct medical costs to households and community is likely.