bluuebunny/crossref_metadata_2025_split huggingface dataset:
The NPK1 mitogen‐activated protein kinase kinase kinase contains a functional nuclear localization signal at the binding site for the NACK1 kinesin‐like protein
Authors: Ishikawa Masaki , Soyano Takashi , Nishihama Ryuichi , Machida Yasunori 
Published: March 2003

SummaryThe tobacco mitogen‐activated protein kinase kinase kinase NPK1 localizes to the equatorial region of phragmoplasts by interacting with kinesin‐like protein NACK1. This leads to activation of NPK1 kinase at late M phase, which is necessary for cell plate formation. Until now, its localization during interphase has not been reported. We investigated the subcellular localization of NPK1 in tobacco‐cultured BY‐2 cells at interphase using indirect immunofluorescence microscopy and fusion to green fluorescent protein (GFP). Fluorescence of anti‐NPK1 antibodies and GFP‐fused NPK1 were detected only in the nuclei of BY‐2 cells at interphase. Examination of the amino acid sequence of NPK1 showed that at the carboxyl‐terminal region in the regulatory domain, which contains the binding site of NACK1, NPK1 contained a cluster of basic amino acids that resemble a bipartite nuclear localization signal (NLS). Amino acid substitution mutations in the critical residues in putative NLS caused a marked reduction in nuclear localization of NPK1 in BY‐2 cells, indicating that this sequence is functional in tobacco BY‐2 cells. We also found that the 64‐amino acid sequence at the carboxyl terminus that contains NLS sequence is essential for interaction with NACK1, and that mutations in the NLS sequence prevented NPK1 from interacting with NACK1. Thus, the amino acid sequence at the carboxyl‐terminal region of NPK1 has dual functions for nuclear localization during interphase and binding NACK1 in M phase.
Evidence that the Arabidopsis nuclear gibberellin signalling protein GAI is not destabilised by gibberellin
Authors: Fleck Barbara , Harberd Nicholas P. 
Published: March 2003

SummaryPlant growth is regulated by bioactive gibberellin (GA), although there is an unexplained diversity in the magnitude of the GA responses exhibited by different plant species. GA acts via a group of orthologous proteins known as the DELLA proteins. The Arabidopsis genome contains genes encoding five different DELLA proteins, the best known of which are GAI and RGA. The DELLA proteins are thought to act as repressors of GA‐regulated processes, whilst GA is thought to act as a negative regulator of DELLA protein function. Recent experiments have shown that GA induces rapid disappearance of nuclear RGA, SLR1 and SLN1 (DELLA proteins from rice and barley), suggesting that GA signalling and degradation of DELLA proteins are coupled. However, RGL1, another Arabidopsis DELLA protein, does not disappear from the nucleus in response to GA treatment. Here, we present evidence suggesting that GAI, like RGL1, is stable in response to GA treatment, and show that transgenic Arabidopsis plants containing constructs that enable high‐level expression of GAI exhibit a dwarf, GA non‐responsive phenotype. Thus, GAI appears to be less affected by GA than RGA, SLR1 or SLN1. We also show that neither of the two putative nuclear localisation signals contained in DELLA proteins are individually necessary for nuclear localisation of GAI. The various DELLA proteins have different properties, and we suggest that this functional diversity may explain, at least in part, why plant species differ widely in their GA response magnitudes.
Inverted repeat of a heterologous 3′‐untranslated region for high‐efficiency, high‐throughput gene silencing
Authors: Brummell David A. , Balint‐Kurti Peter J. , Harpster Mark H. , Palys Joseph M. , Oeller Paul W. , Gutterson Neal 
Published: March 2003

SummaryThis report describes a method for the easy generation of inverted repeat constructs for the silencing of genes of unknown sequence which is applicable to high‐throughput studies. This improved procedure for high‐efficiency gene silencing is specific for a target gene, but does not require inverted repeat DNA of the target gene in the construct. The method employs an inverted repeat of the 3′‐untranslated region (3′‐UTR) of a heterologous gene, and has been demonstrated using the 3′‐UTR region of the nopaline synthase (nos) gene from Agrobacterium tumefaciens, which is often used as the 3′‐UTR for transgene constructs. In a population of independent tomato primary transformants harboring a stably integrated polygalacturonase (PG) transgene driven by a constitutive promoter and linked to an inverted repeat of the nos 3′‐UTR, 51 of 56 primary transformants (91% of the population) showed highly effective post‐transcriptional silencing of the PG gene, with PG mRNA abundance in ripe fruit reduced by 98% or more. The method was also effective in Arabidopsis, where two different, relatively uncharacterized plant transcription factors were also targeted effectively. This method has the advantage of ease and rapidity in preparation of the constructs, since a gene of interest can be inserted into a binary vector already containing the promoter and the inverted nos domain in a single‐cloning step, and does not require any knowledge of the DNA sequence. The approach is suitable for high‐throughput gene silencing studies, where it is necessary to investigate the function of hundreds to thousands of uncharacterized genes.
Function of a mitogen‐activated protein kinase pathway in N gene‐mediated resistance in tobacco
Authors: Jin Hailing , Liu Yidong , Yang Kwang‐Yeol , Kim Cha Young , Baker Barbara , Zhang Shuqun 
Published: March 2003

SummaryThe active defense of plants against pathogens often includes rapid and localized cell death known as hypersensitive response (HR). Protein phosphorylation and dephosphorylation are implicated in this event based on studies using protein kinase and phosphatase inhibitors. Recent transient gain‐of‐function studies demonstrated that the activation of salicylic acid‐induced protein kinase (SIPK) and wounding‐induced protein kinase (WIPK), two tobacco mitogen‐activated protein kinases (MAPKs) by their upstream MAPK kinase (MAPKK), NtMEK2 leads to HR‐like cell death. Here, we report that the conserved kinase interaction motif (KIM) in MAPKKs is required for NtMEK2 function. Mutation of the conserved basic amino acids in this motif, or the deletion of N‐terminal 64 amino acids containing this motif significantly compromised or abolished the ability of NtMEK2DD to activate SIPK/WIPK in vivo. These mutants were also defective in interacting with SIPK and WIPK, suggesting protein–protein interaction is required for the functional integrity of this MAPK cascade. To eliminate Agrobacterium that is known to activate a number of defense responses in transient transformation experiments, we generated permanent transgenic plants. Induction of NtMEK2DD expression by dexamethasone induced HR‐like cell death in both T1 and T2 plants. In addition, by using PVX‐induced gene silencing, we demonstrated that the suppression of all three known components in the NtMEK2–SIPK/WIPK pathway attenuated N gene‐mediated TMV resistance. Together with previous report that SIPK and WIPK are activated by TMV in a gene‐for‐gene‐dependent manner, we conclude that NtMEK2–SIPK/WIPK pathway plays a positive role in N gene‐mediated resistance, possibly through regulating HR cell death.
The risk debate in blood transfusion: how perceptions, beliefs and behaviours can be shaped by an efficient communication
Authors: Hossenlopp C. 
Published: March 2003

The issues of risk and safety have been of prime importance in our field over the last decade. The profession has considered the question of risk assessment and risk management in depth, concentrating first and foremost on technical (biological) approaches, a little less on economy and least of all on the social aspect of the question.Transfusion therapy is probably safer than it has ever been, but public perception and the increase in litigations may suggest the opposite. Transfusion professionals must re‐inspire confidence in the field.In transfusion medicine, informing the patient about to receive blood products is part of the fundamental relationship that associates the duties of the physician and the patient's rights.It has been shown that the basic principles of the communication process constitute a transfer of information. In order to be effective, the communication process should include a feedback, i.e. dialogue between the sender and the receiver of the information.This paper first introduces the notion of risk and the theories about its perception. It then presents the results of a study carried out in a French hospital, aiming to evaluate the effectiveness of the communication process between patients about to receive blood products and the physician/surgeon in charge of the patients. The discussion considers how efficient communication can shape the perception of risks and make blood transfusion risks ‘socially acceptable’.
Intervention to promote appropriate blood use in India
Authors: Bray T. J. , Salil P. , Weiss H. A. , Porter J. D. H. 
Published: March 2003

Summary The decision to transfuse should be guided by information on the risks and benefits of transfusion. Safer alternatives should be considered. Properly screened blood or components from a reputable source should be used. In this study, a simple, self‐educating transfusion request form was developed, and its effects on transfusion practice were assessed, using a cluster‐randomized trial. Transfusions at six study hospitals and six control hospitals at four locations in India (Delhi, Bangalore, Nasik and Imphal) were monitored over a 4‐month pre‐intervention period and a 5‐month post‐intervention period. During the trial, 56 171 units were transfused to in‐patients at the participating hospitals. Among the six intervention hospitals, there was some evidence of a nonsignificant post‐intervention reduction in all the three main outcome measures: number of transfusion requests per admission (P = 0·09), number of units transfused per admission (P = 0·11) and number of crossmatches per admission (P = 0·06). No such changes were seen at control sites over the same period.Simple interventions to promote good clinical practice can have an effect, but may be better placed within longer term, broad‐based strategies that are able to consider some of the background factors. Lack of clinical training, the hospital environment and fragmented blood bank services influence the way blood is used in India. It is thought that the intervention was ultimately unsuccessful because these factors remained as detrimental influences. A focus on education, policy and infrastructure in line with the new National Blood Policy will be important in coming years.
Quinine pharmacokinetics: ototoxic and cardiotoxic effects in healthy Caucasian subjects and in patients with falciparum malaria
Authors: Claessen  , Van Boxtel  , Perenboom  , Tange  , Wetsteijn  , Kager  
Published: March 2003

Summary
objective To study the pharmacokinetic behaviour of quinine in Caucasians with and without malaria. method Quinine‐dihydrochloride was administered intravenously as a single dose of 300 mg to 12 healthy subjects and as multiple doses of 600 mg in 4 h every 8 h in 10 patients with falciparum malaria. Plasma quinine concentrations were measured by high‐performance liquid chromatography. results Quinine pharmacokinetics are time‐dependent: the apparent elimination halftime is shorter in the accumulation phase than in the elimination phase; in malaria patients the maximal quinine concentration was reached in half the time calculated on the basis of the elimination phase after the last quinine infusion. Nevertheless a loading dose seemed advisable to reach adequate therapeutic levels quickly. In malaria patients the highest plasma concentrations during or at the end of the infusions were positively correlated with body weight. There was no correlation between body weight and the volume of distribution of quinine as calculated during the elimination phase. Hearing loss was audiometrically documented in 9 healthy subjects at a mean maximal plasma quinine concentration of only 2 mg/l. All malaria patients suffered serious cochlear hearing impairment. The ototoxic effects in both healthy subjects and patients appeared to be reversible. No electrographic changes were noted in the healthy subjects, whereas a clinically insignificant mean lengthening of the corrected QT interval was seen in the malaria patients. conclusion Intravenous quinine pharmacokinetics in healthy Caucasians were similar to those reported for Nigerian or Thai subjects. At effective doses quinine causes considerable but reversible cochlear hearing losses in both healthy persons and in patients. Our findings do support the need for a loading dose. The fact that in malaria patients there was no correlation between body weight and quinine VD as calculated during the elimination phase renders questionable the usefulness of dosing quinine according to body weight.
In vitro antiplasmodial activity of Central American medicinal plants
Authors: Jenett‐Siems Kristina , Mockenhaupt Frank P. , Bienzle Ulrich , Gupta Mahabir P. , Eich Eckart 
Published: March 2003

Summary The in vitro antiplasmodial activities of 14 plant species traditionally used in Central America for the treatment of malaria or fever were evaluated. Lipophilic extracts of Piper hispidum, Siparuna andina, S. pauciflora, S. tonduziana, and Xylopia cf. frutescens, proved to be active against both a chloroquine‐sensitive and a resistant strain of Plasmodium falciparum. IC50 values ranged between 3.0 μg/ml and 21.9 μg/ml; however, moderate cytotoxicity of active extracts was observed. Bioactivity‐guided fractionation of Piper hispidum yielded 2′,4,6′‐trihydroxy‐4′‐methoxydihydrochalcone (asebogenin) as an active compound.
Maintenance and sustained use of insecticide‐treated bednets and curtains three years after a controlled trial in western Kenya*
Authors: Kachur S. Patrick , Phillips‐Howard Penelope A. , Odhacha Amos M. , Ruebush Trenton K. , Oloo Aggrey J. , Nahlen Bernard L. 
Published: March 2003

Summary In large experimental trials throughout Africa, insecticide‐treated bednets and curtains have reduced child mortality in malaria‐endemic communities by 15%–30%. While few questions remain about the efficacy of this intervention, operational issues around how to implement and sustain insecticide‐treated materials (ITM) projects need attention. We revisited the site of a small‐scale ITM intervention trial, 3 years after the project ended, to assess how local attitudes and practices had changed. Qualitative and quantitative methods, including 16 focus group discussions and a household survey (n = 60), were employed to assess use, maintenance, retreatment and perceptions of ITM and the insecticide in former study communities. Families that had been issued bednets were more likely to have kept and maintained them and valued bednets more highly than those who had been issued curtains. While most households retained their original bednets, none had treated them with insecticide since the intervention trial was completed 3 years earlier. Most of those who had been issued bednets repaired them, but none acquired new or replacement nets. In contrast, households that had been issued insecticide‐treated curtains often removed them. Three (15%) of the households issued curtains had purchased one or more bednets since the study ended. In households where bednets had been issued, children 10 years of age and younger were a third as likely to sleep under a net as were adults (relative risk (RR) = 0.32; 95% confidence interval (95%CI) = 0.19, 0.53). Understanding how and why optimal ITM use declined following this small‐scale intervention trial can suggest measures that may improve the sustainability of current and future ITM efforts.
Prevalence and distribution of soil‐transmitted helminth (STH) infections in urban and indigenous schoolchildren in Ortigueira, State of Paranà, Brasil: implications for control
Authors: Scolari Carla , Torti Carlo , Beltrame Anna , Matteelli Alberto , Castelli Francesco , Gulletta Maurizio , Ribas Milton , Morana Serenella , Urbani Carlo 
Published: March 2003

Summary Soil‐transmitted helminth (STH) infections represent a major public health problem in poor and developing countries. During the period September‐October 1998 we conducted an epidemiological survey of STH infections in schoolchildren of an urban area (group A) and an indigenous reserve (group B), in the Municipality of Ortigueira, State of Paranà, Brazil, to assess potential benefits of mass treatment. Stool samples were examined for helminth eggs by quantitative (Kato‐Katz) technique to determine the prevalence and intensity of intestinal parasitic infection. Moreover, we examined the relationship between prevalence and intensity of STH infections and housing/hygienic factors (by means of a 7‐item questionnaire). 236 schoolchildren aged 5–15 years were enrolled, 136 in group A and 100 in group B. The prevalence of STH infections was significantly higher in group B (93%) than in group A (22%) (P < 0.001). Detected parasites were: A. lumbricoides (16.1% prevalence in group A, 88% in group B, P < 0.001), hookworms (5.8% in group A, 52% in group B, P < 0.001) and T. trichiura (5.1% in group A, 2% in group B, P= 0.2). Heavy infections were detected in 2.9% and 23% of the children in group A and B, respectively (P < 0.001). Housing/hygienic indicators were significantly poorer in group B. A statistically significant correlation was observed between total prevalence of STH infections and prevalence of high‐intensity infections with most housing/hygienic variables. On the basis of these results, mass treatment and educational interventions were suggested for the indigenous community, whereas target treatment and educational interventions were suggested for the urban community. Even in a geographically homogeneous area different epidemiological realities can be found, which in turn can influence infection levels and control programmes.
Rapid therapeutic response onset of a new pharmaceutical form of chloroquine phosphate 300 mg: effervescent tablets
Authors: Yanze Maximun Frédéric , Duru Christian , Jacob Maurice , Bastide Jean Marie , Lankeuh Marguerite 
Published: March 2003

OBJECTIVE To compare the efficiency, safety and taste of two pharmaceutical forms of chloroquine phosphate 300 mg: effervescent tablets against uncoated tablets. METHOD An open randomized study with 60 adults who suffered from acute uncomplicated Plasmodium falciparum malaria in three health centres in Nkongsamba health district, Cameroon. RESULTS Mean times to fever clearance, symptoms clearance and asexual parasites clearance were longer in the uncoated tablets group: 36 h (range 24–48 h, SD=16.8) vs. 60 h (range 24–96 h, SD=31.2, P=0.001) for fever clearance, 36 h (24–48 h, SD=16.8) vs. 48 h (24–72, SD=24, P=0.001) for symptoms clearance and 48 h (24–72, SD=1) vs. 72 h (48–96, SD=24, P=0.001) for parasitaemia clearance. Uncoated tablets took significantly longer to achieve 50% reduction of the initial asexual parasite density: (mean/SD) 19.2 h/7 vs. 52.8 h/16.8, P < 0.00001. The adverse effects in the two groups were similar, P > 0.05. The cure rate at day 7 in the two groups was similar, P > 0.05. There was no chloroquine resistance in the effervescent tablets group but one RI and one RII resistance in the uncoated tablets group. The taste of the two pharmaceutical forms was significantly different, P < 0.00001. Effervescent tablets tasted sweet (score=7.93), whereas uncoated tablets were bitter (score=2.07). CONCLUSION Effervescent tablets of chloroquine phosphate 300 mg work faster than uncoated tablets and because of their safe use and sweet taste achieve good therapeutic compliance.
Evaluation of staff performance and material resources for integrated schistosomiasis control in Northern Senegal
Authors: Van Der Werf Marieke J. , Mbaye Amadou , Sow Seydou , Gryseels Bruno , De Vlas Sake J. 
Published: March 2003

BACKGROUND A project to improve integrated control of schistosomiasis in the primary health care system of Northern Senegal was implemented from February 1995 until September 1999, shortly after a Schistosoma mansoni outbreak. The activities included additional training of doctors and nurses in symptom‐based treatment and making praziquantel (PZQ) available for an affordable price. OBJECTIVE To investigate staff performance and the availability and costs of diagnostic materials and PZQ at the end of this intervention project. METHODS We performed structured interviews with staff from 55 health care facilities in five districts. RESULTS Respondents from 23 health care facilities reported both S. haematobium and S. mansoni in the coverage area, 32 reported only S. haematobium and three only S. mansoni. The average cost to patients for consultation, diagnosis, treatment and transportation to a referral health care facility was approximately 1.60 Euro. Fifty‐seven per cent of the health care facilities with reported S. haematobium in the coverage area treated patients presenting with haematuria on symptoms; 56% of the health care facilities with reported S. mansoni in the coverage area treated patients presenting with blood in stool on symptoms. Thirteen per cent performed a diagnostic test for patients presenting with haematuria and 12% for patients presenting with blood in stool. The remainder, approximately one‐third of the health care facilities, referred their patients to another facility for a diagnostic test. Implementation of symptom‐based treatment in all health care facilities will reduce the total costs by 0.43 Euro (29%) for patients infected with S. haematobium and 0.78 Euro (46%) for patients infected with S. mansoni. Of the 53 health care facilities with schistosomiasis in their area, 37 had PZQ in stock of which 33 (88%) sold PZQ for the recommended retail price of 0.15 Euro per tablet (or 0.60 Euro per course of four tablets) or lower. CONCLUSION Four years after the start of the intervention project, patients presenting with schistosomiasis related symptoms can generally expect proper diagnosis and treatment at all levels of the health care system in Northern Senegal, either at the initial visited health care facility or after referral. However, a further reduction of the total costs of treatment is still possible by a better implementation of symptom‐based treatment and further reduction of the costs of PZQ.
Examining out‐of‐pocket expenditure on health care in Nouna, Burkina Faso: implications for health policy
Authors: Mugisha Frederick , Kouyate Bocar , Gbangou Adjima , Sauerborn Rainer 
Published: March 2003

OBJECTIVE To examine household out‐of‐pocket expenditure on health care, particularly malaria treatment, in rural Burkina Faso. METHOD Comprehensive analysis of out‐of‐pocket expenditure on health care through a descriptive analysis and a second, multivariate analysis using the Tobit model with emphasis on malaria, based on 800 urban and rural households in Nouna health district. RESULTS Households will spend less on malaria, either in or outside the health facility, if given the choice to do so, because they feel confident to self‐treat malaria. Seeking health care from a qualified health worker incurs more out‐of‐pocket expenditure than self‐treatment and traditional healers, and if necessary, households sell off assets to offset the expenditure. More than 80% of household out‐of‐pocket expenditure is allocated to drugs. CONCLUSION This has policy implications for malaria control and the Roll Back Malaria Initiative. Communities need to be educated on the risks of malaria complications and the potential risk of inappropriate diagnosis and treatment. Drug or health services pricing policy needs to create an incentive to use the health services. In the fight against malaria, building alliances between households, traditional healers and health workers is essential.