bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular phylogeny and evolution of host‐plant use in conifer seed chalcids in the genus Megastigmus (Hymenoptera: Torymidae)
Authors: Auger‐Rozenberg Marie‐Anne , Kerdelhué Carole , Magnoux Emmanuelle , Turgeon Jean , Rasplus Jean‐Yves , Roques Alain
Published: August 2005

Abstract.  Phylogenetic relationships amongst Megastigmus species (Chalcidoidea: Torymidae) associated with conifer seeds were inferred from DNA sequence data. Twenty‐nine species of seed chalcids were analysed using two different genes, cytochrome b (mitochondrial DNA) and the D2 domain of the 28S ribosomal DNA. Maximum‐parsimony and maximum‐likelihood analyses showed that taxa formed two monophyletic groups, one clade comprising all species associated with Cupressaceae and Taxodiaceae hosts with the exception of Chamaecyparis, and the other clade composed of species associated with Pinaceae. Species infesting Cupressaceae and Taxodiaceae seemed to be specialized to particular host genera or even to be species specific, which was consistent with a taxonomic radiation following initial host adaptation. By contrast, Megastigmus species associated with Pinaceae appeared capable of shifting onto different congeneric species or even onto a new host genus, with their evolution apparently less constrained by plant association. We hypothesized that the Megastigmus group associated with Pinaceae may have a much higher invasive potential than that related to Cupressaceae. The study also confirmed the presence of invasive Nearctic species in the Palaearctic, and demonstrated the existence of a cryptic species complex.
Steady‐state transposon mutagenesis in inbred maize
Authors: McCarty Donald R. , Mark Settles Andrew , Suzuki Masaharu , Tan Bao Cai , Latshaw Susan , Porch Tim , Robin Kevin , Baier John , Avigne Wayne , Lai Jinsheng , Messing Joachim , Koch Karen E. , Curtis Hannah L.
Published: August 2005

We implement a novel strategy for harnessing the power of high‐copy transposons for functional analysis of the maize genome, and report behavioral features of the Mutator system in a uniform inbred background. The unique UniformMu population and database facilitate high‐throughput molecular analysis of Mu‐tagged mutants and gene knockouts. Key features of the population include: (i) high mutation frequencies (7% independent seed mutations) and moderation of copy number (approximately 57 total Mu elements; 1–2 MuDR copies per plant) were maintained by continuous back‐crossing into a phenotypically uniform inbred background; (ii) a bz1‐mum9 marker enabled selection of stable lines (loss of MuDR), inhibiting further transpositions in lines selected for molecular analysis; (iii) build‐up of mutation load was prevented by screening Mu‐active parents to exclude plants carrying pre‐existing seed mutations. To create a database of genomic sequences flanking Mu insertions, selected mutant lines were analyzed by sequencing of MuTAIL PCR clone libraries. These sequences were annotated and clustered to facilitate bioinformatic subtraction of ancestral elements and identification of insertions unique to mutant lines. New insertions targeted low‐copy, gene‐rich sequences, and in silico mapping revealed a random distribution of insertions over the genome. Our results indicate that Mu populations differ markedly in the occurrence of Mu insertion hotspots and the frequency of suppressible mutations. We suggest that controlled MuDR copy number in UniformMu lines is a key determinant of these differences. The public database (http://uniformmu.org; http://endosperm.info) includes pedigree and phenotypic data for over 2000 independent seed mutants selected from a population of 31 548 F2 lines and integrated with analyses of 34 255 MuTAIL sequences.
Construction and screening of subtracted cDNA libraries from limited populations of plant cells: a comparative analysis of gene expression between maize egg cells and central cells
Authors: Lê Quyên , Gutièrrez‐Marcos José F. , Costa Liliana M. , Meyer Stephanie , Dickinson Hugh G. , Lörz Horst , Kranz Erhard , Scholten Stefan
Published: August 2005

The analysis of cell type‐specific gene expression is an essential step in understanding certain biological processes during plant development, such as differentiation. Although methods for isolating specific cell types have been established, the application of cDNA subtraction to small populations of isolated cell types for direct identification of specific or differentially expressed transcripts has not yet been reported. As a first step in the identification of genes expressed differentially between maize egg cells and central cells, we have manually isolated these types of cell, and applied a suppression‐subtractive hybridization (SSH) strategy. After microarray screening of 1030 cDNAs obtained from the subtracted libraries, we identified 340 differentially expressed clones. Of these, 142 were sequenced, which resulted in the identification of 62 individual cDNAs. The expression patterns of 20 cDNAs were validated by quantitative RT–PCR, through which we identified five transcripts with cell type‐specific expression. The specific localization of some of these transcripts was also confirmed by in situ hybridization on embryo sac sections. Taken together, our data demonstrate the effectiveness of our approach in identifying differentially expressed and cell type‐specific transcripts of relatively low abundance. This was also confirmed by the identification of previously reported egg cell‐ and central cell‐specific genes in our screen. Importantly, from our analysis we identified a significant number of novel sequences not present in other embryo sac or, indeed, in other plant expressed sequence tag (EST) databases. Thus, in combination with standard EST sequencing and microarray hybridization strategies, our approach of differentially screening subtracted cDNAs will add substantially to the expression information in spatially highly resolved transcriptome analyses.
Virus‐induced gene silencing is an effective tool for assaying gene function in the basal eudicot speciesPapaver somniferum(opium poppy)
Authors: Hileman Lena C. , Drea Sinéad , de Martino Gemma , Litt Amy , Irish Vivian F.
Published: August 2005

Virus‐induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are resistant to conventional genetic approaches. However, VIGS has been shown to be effective in only a few, closely related plant species. Tobacco rattle virus (TRV), a bipartite RNA virus, has a wide host range and so in principle could serve as an efficient vector for VIGS in a diverse array of plant species. Here we show that a vector based on TRV sequences is effective at silencing the endogenousphytoene desaturase(PapsPDS) gene inPapaver somniferum(opium poppy). We show that this vector does not compromise the growth or reproduction of poppy and the plants did not display viral symptoms. The silencing ofPapsPDSresulted in a significant reduction inPapsPDSmRNA and a concomitant photobleached phenotype. The ability to rapidly assay gene function inP. somniferumwill be valuable in manipulation of the opiate pathway in this pharmaceutically important species. We suggest that our vacuum infiltration method used to deliver TRV‐based vectors into poppy is a promising approach for expanding VIGS to diverse angiosperm species in which traditional delivery methods fail to induce VIGS. Furthermore, these studies demonstrate the utility of TRV‐VIGS for probing gene function in a basal eudicot species that is phylogenetically distant from model plant species.
Albendazole for the control and elimination of lymphatic filariasis: systematic review
Authors: Critchley Julia , Addiss David , Ejere Henry , Gamble Carrol , Garner Paul , Gelband Hellen ,   the International Filariasis Review Group
Published: August 2005

Objectives  The Global Programme to Eliminate Lymphatic Filariasis recommends albendazole in combination with other antifilarial drugs. This systematic review examines albendazole in treatment and control of lymphatic filariasis.

Datasources  The Cochrane Controlled Trials Register, MEDLINE and EMBASE to April 2005; contacting experts, international organisations and drug manufacturers.

Methods  Randomised or quasi‐randomised controlled trials included; two reviewers independently assessed eligibility, quality, and extracted data. We calculated the relative risk of microfilaraemia (mf) prevalence using fixed effect, or random effects model in case of heterogeneity.

Results  Six trials met inclusion criteria. Three trials compared albendazole with placebo: no effect was demonstrated on mf prevalence, but density was lower in one of the three studies at 6 months. Three trials added albendazole to ivermectin, with no demonstrable effect; prevalence tended to be lower at 4–6 months but not at 12 months (4–6 months; RR 0.49, 95% CI 0.18 to 1.39, n = 255, 2 trials; 12 months: RR 1.00, 95% CI 0.88 to 1.13, n = 348, 2 trials). Mf density was significantly lower in two of the three trials; one of two trials measuring density at 12 months showed a difference. Three trials added albendazole to diethylcarbamazine; two were small trials with no difference demonstrated; the third study tended to favour combination at 6 months (RR = 0.62, 95% CI 0.32 to 1.21, n = 491), with a significant difference for density.

Conclusions  The effect of albendazole against adult and larval filarial parasites, alone and in combination with other antifilarial drugs, deserves further rigorous research.
Unaffordable or cost‐effective?: introducing an emergency referral system in rural Niger
Authors: Bossyns Paul , Abache Ranaou , Abdoulaye Mahaman Sani , Lerberghe Wim Van
Published: August 2005

Objectives  An important investment was made in two health districts in Niger to organize an emergency referral system. This study estimates its impact and cost‐effectiveness in relation with external determinants.

Methods  After installing a solar radio network in the health centres, emergency calls and related data were monitored over 7 years and investment and recurrent costs for the system were estimated.

Results  The number of emergency calls increased significantly in both districts. In 2003, the total yearly cost for the district amounted to US$ 14 147, the cost per useful and successful call was US$ 49 and the cost per inhabitant and per year was about US$ 0.06.

Conclusion  The impressive and immediate impact on the health system, the relatively low recurrent cost and the minimal management requirements for the health service make the investment very worthwhile. Organizing emergency evacuation systems should be a priority for any health district in the world.
Control of mucocutaneous leishmaniasis, a neglected disease: results of a control programme in Satipo Province, Peru
Authors: Guthmann Jean‐Paul , Arlt Doris , Garcia Luis Miguel Leon , Rosales Milena , de Jesus Sanchez Juan , Alvarez Eugenia , Lonlas Sylvaine , Conte Mado , Bertoletti Guillermo , Fournier Christophe , Huari Roberto , Torreele Els , Llanos‐Cuentas Alejandro
Published: August 2005

Mucocutaneous leishmaniasis (MCL) is an important health problem in many rural areas of Latin America, but there are few data on the results of programmatic approaches to control the disease. We report the results of a control programme in San Martin de Pangoa District, which reports one of the highest prevalences of MCL in Peru. For 2 years (2001–2002), the technicians at the health post were trained in patient case management, received medical support and were supplied with antimonials. An evaluation after 2 years showed the following main achievements: better diagnosis of patients, who were confirmed by microscopy in 34% (82/240) of the cases in 2001 and 60% of the cases (153/254) in 2002; improved follow‐up during treatment: 237 of 263 (90%) patients who initiated an antimonial therapy ended the full treatment course; improved follow‐up after treatment: 143 of 237 (60%) patients who ended their full treatment were correctly monitored during the required period of 6 (cutaneous cases) or 12 (mucosal cases) months after the end of treatment. These achievements were largely due to the human and logistical resources made available, the constant availability of medications and the close collaboration between the Ministry of Health, a national research institute and an international non‐governmental organization. At the end of this period, the health authorities decided to register a generic brand of sodium stibogluconate, which is now in use. This should allow the treatment of a significant number of additional patients, while saving money to invest in other facets of the case management.
Paying for health in two Rwandan provinces: financial flows and flaws
Authors: Kalk Andreas , Mayindo Jean Kagubare , Musango Laurent , Foulon Gerard
Published: August 2005

The study intended to analyse the financial flows in two provinces within the Rwandan health system through the review of all available documentation and through interviews with key informants, to assess the scope for improved resource allocation. In Rwanda, there exists a large deficit of available financial resources in the health sector in general, and more specifically at health centre level. To improve this situation, it is considered to cover a large proportion of the entire population by mutual health insurance schemes. The schemes are able to pool certain risks, and they definitely improve financial access to health services. Nonetheless, they are inaccessible to the ‘very poor’, and – due to their limited financial base – they are unable to cover a complementary health care package. It is unlikely that they will mobilize substantial additional resources for health. External long‐term commitments are required to cover this gap. A reassurance and readjustment system between the various insurance schemes should be established in order to increase financial protection provided. It might link up with insurance schemes in the formal employment sector. The combination of such a support for health insurance with performance‐related incentives for health staff has the potential to increase both equity and quality of health services simultaneously and substantially.
Roll Back Malaria in the aftermath of complex emergencies: the example of Afghanistan
Authors: Kolaczinski Jan
Published: August 2005

The Roll Back Malaria (RBM) partnership was established in 1998 to address the worldwide malaria burden through a new approach. It was founded by four multilateral agencies that were already committed to supporting malaria control: the United Nations Development Programme, the United Nations Children's Fund, the World Bank and the World Health Organization. In contrast to previous campaigns, RBM focuses on strengthening existing health services to ensure that malaria can be controlled through an integrated approach, rather than through creation and maintenance of a separate structure. Field experience of RBM is rarely documented in the open scientific literature, making it difficult to judge its success and apply lessons learned to other settings. This paper aims at improving on this by documenting experience of RBM support in Afghanistan during initial post‐conflict reconstruction. From the analysis some recommendations emerged: (i) technical in‐country support needs to be provided for a number of years, as short postings are likely to provide only temporary solutions and may fail to contribute towards medium or long‐term development; (ii) the role and responsibilities of support staff need to be clear to all RBM partners; (iii) part of this role should be to act as focal point, in an attempt to improve on coordination and (iv) if possible, support staff should be seconded to the MoH, to avoid association of the RBM initiative with a specific international organization and to further the multi‐sectorial partnership approach.
Quantifying the level of under‐detection of Trypanosoma brucei rhodesiense sleeping sickness cases
Authors: Odiit M. , Coleman P. G. , Liu W.‐C. , McDermott J. J. , Fèvre E. M. , Welburn S. C. , Woolhouse M. E. J.
Published: August 2005

To formally quantify the level of under‐detection of Trypanosoma brucei rhodesiense sleeping sickness (SS) during an epidemic in Uganda, a decision tree (under‐detection) model was developed; concurrently, to quantify the subset of undetected cases that sought health care but were not diagnosed, a deterministic (subset) model was developed. The values of the under‐detection model parameters were estimated from previously published records of the duration of symptoms prior to presentation and the ratio of early to late stage cases in 760 SS patients presenting at LIRI hospital, Tororo, Uganda during the 1988–1990 epidemic of SS. For the observed early to late stage ratio of 0.47, we estimate that the proportion of under‐detection in the catchment area of LIRI hospital was 0.39 (95% CI 0.37–0.41) i.e. 39% of cases are not reported. Based on this value, it is calculated that for every one reported death of SS, 12.0 (95% CI 11.0–13.0) deaths went undetected in the LIRI hospital catchment area – i.e. 92% of deaths are not reported. The deterministic (subset) model structured on the possible routes of a SS infection to either diagnosis or death through the health system or out of it, showed that of a total of 73 undetected deaths, 62 (CI 60–64) (85%) entered the healthcare system but were not diagnosed, and 11 (CI 11–12) died without seeking health care from a recognized health unit. The measure of early to late stage presentation provides a tractable measure to determine the level of rhodesiense SS under‐detection and to gauge the effects of interventions aimed at increasing treatment coverage.
Plasmodium falciparum dhfr but not dhps mutations associated with sulphadoxine‐pyrimethamine treatment failure and gametocyte carriage in northern Ghana
Authors: Mockenhaupt Frank P. , Teun Bousema J. , Eggelte Teunis A. , Schreiber Jonas , Ehrhardt Stephan , Wassilew Nasstasja , Otchwemah Rowland N. , Sauerwein Robert W. , Bienzle Ulrich
Published: August 2005

Both use of sulphadoxine‐pyrimethamine (SP) and SP‐resistance of Plasmodium falciparum are increasing in sub‐Saharan Africa. Mutations in the P. falciparum dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) genes can predict treatment failure of SP, however, the degree of this relationship varies regionally. In northern Ghana, pre‐treatment dhfr/dhps genotypes were examined in 126 children and associations with PCR‐corrected SP treatment outcome and gametocyte carriage were analysed. SP treatment failure within 4 weeks of follow‐up occurred in 28%. Among all pre‐treatment isolates, the dhfr triple mutation (Ile‐51 + Arg‐59 + Asn‐108) was detected in 47%. Compared with dhfr wildtype parasites, the presence of the dhfr triple mutation increased the risk of treatment failure tenfold. Likewise, parasite clearance was delayed in the presence of dhfr variants. Dhfr mutants and dhps Gly‐437 were selected in treatment failure isolates. Gametocytaemia 1 week following treatment was strongly associated with dhfr mutations. Remarkably, this was also true for the prevalence of gametocytes at recruitment. Dhps alleles did neither influence treatment outcome nor gametocyte carriage. In northern Ghana, the prevalence of the dhfr triple mutation can be used as a tool to screen for and to monitor SP resistance. The lack of association between dhps alleles and SP treatment outcome suggests a minor role of these molecular markers in this region at present.
Early diagnosis and treatment of uncomplicated malaria and patterns of health seeking in Vietnam
Authors: Giao Phan T. , de Vries Peter J. , Binh Tran Q. , Nam Nguyen V. , Kager Piet A.
Published: August 2005

Early diagnosis and treatment of malaria (EDTM) is a key component of malaria control. The success of EDTM depends on health seeking behaviour and the quality of the health service. This study assessed self‐diagnosis, treatment and treatment delay after the introduction of EDTM in 1993. In southern Vietnam EDTM comprises microscopic diagnosis and free treatment with artemisinin derivatives at public health facilities. Until 2001, 1698 questionnaires had been completed by patients participating in randomized treatment trials of uncomplicated malaria. The presumptive self diagnosis ‘malaria’ increased from 68% in 1993 to 100% in 2001 and self‐treatment decreased, from 74% to 8% in 2000 and 24% in 2001. The median (maximum) delay between first symptoms and seeking treatment at a public health facility decreased from 3 (23) to 1.3 (3) days (P < 0.001) Concomitant was a significant decline of reported incidence of malaria‐associated mortality, severe malaria and uncomplicated malaria. If offered an attractive package of EDTM, patients become sensitized to the possibility of malaria and less likely to self‐treat. EDTM should be provided as soon as possible to all symptomatic patients, aiming at reducing treatment delay to a maximum of 2 days.
Economic evaluation of a policy change from single‐agent treatment for suspected malaria to artesunate‐amodiaquine for microscopically confirmed uncomplicated falciparum malaria in the Oussouye District of south‐western Senegal
Authors: Agnamey P. , Brasseur P. , Cisse M. , Gaye O. , Dumoulin J. , Rigal J. , Taylor W. R. J. , Olliaro P.
Published: August 2005

Senegal is changing policy for case management of uncomplicated falciparum malaria, which hitherto is diagnosed clinically and treated with chloroquine or intramuscular quinine. The WHO recommends artemisinin‐based combinations for treating falciparum malaria, preferably based on a parasitological diagnosis. There are no economic projections if such a policy were introduced in Senegal. We have conducted a preliminary economic assessment of such a policy change. The study took place in the chloroquine‐resistant district of Oussouye in south‐western Senegal. We reviewed clinic registers of the district health posts (n = 5) from 1996 to 2001, and piloted artesunate combined with amodiaquine (at 4 and 10 mg/kg/day × 3 days respectively) (AS‐AQ) for treating slide‐proven falciparum malaria during two rainy seasons (2000 and 2001) at one health centre. These data were used to calculate current direct patient costs (clinic visit, diagnosis, drugs) of malaria treatment and project future costs for the district. The robustness of the model was tested by allowing for different drug failure rates and costs of diagnosis. During 1996–2001, the mean number of primary treatments per year was 7654 for a mean, direct cost of US$17 452 to the community. Clinical diagnosis resulted in over‐treatment: 56% and 66% in the wet and dry seasons respectively. Current policy leads to substantial drug wastage and excess direct costs for the community. The direct costs of implementing AS‐AQ for slide‐proven malaria would be US$8150 (53% less expensive). Studies examining the public health effect and economics of deploying AS‐AQ on a wider scale are underway in Senegal.
Erythroderma in adults: a report of 80 cases
Authors: Rym Benmously Mlika , Mourad Mokni , Bechir Zouari , Dalenda Eleuch , Faika Cherif , Iadh Azaiz Mohamed , Amel Ben Osman‐Dhahri
Published: August 2005

Background  Erythroderma is a generalized erythema of the skin accompanied by a variable degree of scaling. However, most of the published series originate from Western countries. There is only one report from Africa, conducted in Dakar in 1979. The aim of our study was to provide information about this condition in our country.

Patients and methods  Our study was retrospective, dealing with 80 erythrodermic adults, examined between January 1981 and December 2000. Patient information included clinical, laboratory, histopathologic and therapeutic data. We also noted patients’ outcomes within an average follow‐up period of 30 months. Thus, we calculated the mortality rate and estimated the probability of survival without relapses according to the Kaplan‐Meier method.

Results  The frequency of erythroderma in our dermatology department was 0.3. The sex ratio (male/female) was 2.2. The average age of our patients was 53.78 ± 18 years. Previous history of skin disease was found in 49/80 patients (61.2%) and 32 of them were suffering from psoriasis. Hyperthermia was significantly associated with drug reactions (P = 0.013). Hypereosinophilia was not a specific finding. The mortality rate was higher than that of an age‐matched population.

Conclusion  Adult erythroderma is a rare condition. Its most common cause in our study was psoriasis. No cases of atopic dermatitis were found. Prognosis was adversely affected by the presence of erythroderma.
Childhood melanoma: update and treatment
Authors: Huynh Phung M. , Grant‐Kels Jane M. , Grin Caron M.
Published: August 2005

Childhood melanoma is a rare but potentially fatal disease that is important to include in the differential diagnosis of any pigmented lesion in a child. The best prognosis is achieved with early diagnosis and definitive surgical excision. Adjuvant chemotherapy and immunotherapy are options for those with more advanced tumors. Melanoma in children must be treated as aggressively as in adults because childhood melanoma may be equally devastating.
Treatment of keloids and hypertrophic scars with dermojet injections of bleomycin: a preliminary study
Authors: Saray Yasemin , Güleç A. Tülin
Published: August 2005

Background  Numerous treatment modalities have been used to treat keloids and hypertrophic scars, but the optimal treatment has not been established.

Objective  The aim of this study was to determine the efficacy and safety of intralesional jet injection of bleomycin as therapy for keloids and hypertrophic scars that are unresponsive to intralesional steroid injection.

Methods  The study included 14 patients with 15 keloids or hypertrophic scars that had not responded to a minimum of three intralesional injections of triamcinolone acetonide. Multiple jet injections of 0.1 ml of bleomycin (1.5 IU/ml) were administered to each lesion, with injection sites spaced 0.5 mm apart. Injections were repeated each month. Scar height was measured, and scar pliability and erythema were scored at baseline and then monthly during the treatment and follow‐up periods.

Patients’ self‐assessments of subjective symptoms (pruritus and pain) were also scored. Clinical improvement was defined primarily on the basis of scar height reduction (percentage reduction from baseline), and was classified using the following scale: complete flattening (100%), highly significant flattening (> 90%), significant flattening (75–90%), moderate flattening (50–75%), and minimal flattening (< 50%). Pre‐ and post‐treatment mean values for scar height, scar pliability, erythema, pruritus and pain were statistically compared.

Results  The number of sessions required to successfully treat the lesions ranged from two to six. Eleven lesions (73.3%) showed complete flattening, one (6.7%) showed highly significant flattening, two (13.3%) showed significant flattening, and one scar (6.7%) showed moderate flattening. The mean scar height was significantly lower, and the mean scores for scar pliability and erythema were significantly better at the end of treatment (P < 0.001, P < 0.001 and P < 0.001, respectively). The mean scores for pruritus and pain also improved significantly (P < 0.001 and P = 0.01, respectively). The observed side‐effects were hyperpigmentation (four lesions) and skin atrophy (three lesions). No recurrences were noted during follow up (mean duration of 19 months).

Conclusions  Intralesional jet injection of bleomycin is an effective and safe method of treating keloids and hypertrophic scars that are unresponsive to intralesional steroid therapy.
Pityriasis rosea: An important papulosquamous disorder
Authors: González Lenis M. , Allen Robert , Janniger Camila Krysicka , Schwartz Robert A.
Published: August 2005

Pityriasis rosea (PR) is a mild, self‐limited skin disease of unknown etiology. Viral and bacterial causes have been considered as possible causes. Pityriasis rosea typically affects children and young adults. It is characterized by an initial herald patch, followed by the development of salmon‐pink scaly macules which, when localized to the trunk, form along Langer's lines of cleavage. Pityriasis rosea may be difficult to diagnose until the appearance of these characteristic smaller secondary lesions. Several medications can cause a rash similar to PR. Pityriasis rosea must be distinguished from several diseases, including secondary syphilis. The treatment of PR is usually symptomatic.
Poly‐l‐proline type II peptide mimics as probes of the active site occupancy requirements of cGMP‐dependent protein kinase
Authors: Zhang R. , Nickl C. K. , Mamai A. , Flemer S. , Natarajan A. , Dostmann W. R. , Madalengoitia J.S.
Published: August 2005

Abstract:  Based on the X‐ray crystal structure of cAMP‐dependent protein kinase (PKA) with the endogenous inhibitor PKI and the X‐ray crystal structure of cyclin‐dependent kinase 2 (CDK2) with a substrate peptide, a proposal is put forth that some protein kinases bind peptide substrates in their active sites in the poly‐l‐proline type II (PPII) conformation. In this work, PPII peptide mimics are evaluated as pseudosubstrate inhibitors of cGMP‐dependent protein kinase (PKG) to explore if PKG also binds peptide substrates in the PPII conformation. Inhibition data of our PPII mimetics provide evidence that the P − 1, P − 2, and P − 3 residues of substrate peptides bind in the PPII conformation (φ approximately −75°, ψ approximately 145°). In addition, the inhibition data also suggest that the P − 1, P − 2, and P − 3 residues in substrate peptides bind with a gauche(−) χ1 angle.
Effects of anaesthesia using endotracheal intubation or the laryngeal mask airway, or spinal anaesthesia, on oral mucosal host defences during minor surgery
Authors: Lähteenmäki M. , Salo M. , Tenovuo J. , Vahlberg T.
Published: August 2005

Background:  Oral mucosal host defences are altered after anaesthesia and surgery. The effects of endotracheal intubation (ET) and the use of the laryngeal mask airway (LMA) on oral mucosal host defences after minor surgery were compared.

Methods:  Immunological (immunoglobulin A (IgA), IgG and IgM) and non‐immunological (myeloperoxidase, total peroxidase, thiocyanate) oral host defence factors, and amylase and protein concentrations, were measured from saliva pre‐operatively and on the first post‐operative day in 60 ASA I–II patients undergoing minor surgery. The patients underwent general anaesthesia using oral ET or LMA. Spinal anaesthesia (S) was used as control. Serum IgG, IgM and IgA concentrations were determined.

Results:  Protein‐related salivary secretion of amylase and salivary concentration of IgM showed the only statistically significant overall differences between the groups. By contrast, changes were observed within the ET group in the salivary flow rate, protein concentration, amylase activity and immunological host defence factors. Some changes were also observed in the LMA group, but none in the S group. Most non‐immunological test values did not change within any of the groups.

Conclusions:  ET and LMA induced similar oral mucosal host defence responses. There were, however, observations in this study that indicated a stronger response during ET than when LMA was used.
Comparison of alfetanil and ketamine in combination with propofol for patient‐controlled sedation during fiberoptic bronchoscopy
Authors: Hwang J. , Jeon Y. , Park H.‐P. , Lim Y.‐J. , Oh Y.‐S.
Published: August 2005

Background:  During fiberoptic bronchoscopy, propofol, ketamine, benzodiazepines, and opiates are most commonly used, alone or in combination for sedation. The aim of this study was to compare the clinical efficacy of propofol/ketamine with propofol/alfentanil for patient‐controlled sedation (PCS) during fiberoptic bronchoscopy.

Method:  Patients undergoing fiberoptic bronchoscopy were randomly assigned to receive either propofol/alfentanil (PA group; n = 138) or propofol/ketamine (PK group; n = 138) via a patient‐controlled analgesia (PCA) device for sedation and analgesia. Changes in blood pressure, heart rate (HR), and oxygen saturation were monitored. Degree of patient and bronchoscopist satisfaction was evaluated using a 10‐cm visual analog scale (VAS) (0 = extremely uncomfortable to 10 = extremely comfortable).

Results:  After sedation, systolic arterial pressure (SAP) decreased in the PA group, but SAP was stable in the PK group. Compared with values immediately before starting bronchoscopy, SAP and HR increased during the procedure in both groups (P < 0.05). Patients in the PK group showed more satisfaction [(9.5 (6–10) vs. 9.0 (6–10)), P < 0.05] and amnesia (82% vs. 61%, P < 0.01). Despite these differences, the majority (greater than 90%) of the patients in both groups stated that they were comfortable during the procedure.

Conclusion:  Our results show that although both techniques proved effective for sedation in patients undergoing fiberoptic bronchoscopy, ketamine is superior to alfentanil when used in combination with propofol because of the high patient satisfaction and amnesia.
Evaluation of the airway of the SimMan™ full‐scale patient simulator
Authors: Hesselfeldt R. , Kristensen M. S. , Rasmussen L. S.
Published: August 2005

Background: SimMan™ is a full‐scale patient simulator, capable of simulating normal and pathological airways. The performance of SimMan™ has never been critically evaluated.

Methods: Sixty subjects (anesthesiologists, nurse anesthetists, and anesthesia residents) performed mask ventilation, laryngeal mask insertion and endotracheal intubation on SimMan™. The simulator's airway was evaluated using visual analog scales (VAS) and by measuring the subject's performances.

Results: The SimManTMfull‐scale patient simulator's airway is generally acceptably realistic but it significantly differs from the human airway in important aspects. Mask seal was more difficult than in humans whereas Laryngeal mask (LMATM) insertion and function was acceptable. The distance from the teeth to the vallecula was too short. Cervical spine mobility was significantly reduced in the ‘reduced neck movement’ mode but the intubation was only slightly more difficult than in the ‘normal’ mode.

Conclusion: The SimManTMfull‐scale patient simulator's airway is generally acceptably realistic but it significantly differs from the human airway in important aspects. The user must be aware of these aspects in order to obtain maximum benefit from training and evaluation scenarios and when using the simulator for testing new equipment and techniques.
The coagulopathy of massive transfusion
Authors: Hardy J.‐F. , De Moerloose P. , Samama C. M.
Published: August 2005

Recently, the Groupe d’Intérêt en Hémostase Périopératoire reviewed the pathophysiology of coagulopathy in massively transfused, adult and previously haemostatically competent patients in both elective surgical and trauma settings. In this article, we focus on our main observations.

First, in most cases, the onset and severity of coagulopathy associated with massive transfusion differs depending on whether haemorrhage occurs as a result of trauma or elective surgery. In trauma patients, tissue trauma is uncontrolled, the interval between haemorrhage and treatment can vary widely, hypovolemia, shock and hypothermia are frequent, and coagulopathy is often related to the development of disseminated intravascular coagulation. Monitoring of haemostasis occurs late, when coagulopathy is installed, and treatment can be very difficult. In elective surgery patients, the situation remains controlled and, in most cases, a decrease in fibrinogen concentration is observed initially while thrombocytopenia is a late occurrence. Monitoring of haemostasis is ongoing and treatment is usually much simpler.

Second, blood products have changed over time and this has affected the management of the bleeding patient. Contrary to the recommendations of studies published at a time when whole blood was readily available, the first line of treatment (at least in elective surgery patients) ought to be with fresh‐frozen plasma to correct decreased levels of coagulation factors. The role of recombinant activated factor VII to treat bleeding that cannot be controlled by conventional measures remains to be clarified.

Coagulopathy associated with massive transfusion remains an important clinical problem. Treatment strategies must be adapted to the context and to the blood products available. Nevertheless, the level of evidence supporting specific treatment options is low and more studies are required to guide our management of massively transfused patients.
Detection of HCV and HIV‐1 antibody negative infections in Scottish and Northern Ireland blood donations by nucleic acid amplification testing
Authors: Jarvis L. M. , Dow B. C. , Cleland A. , Davidson F. , Lycett C. , Morris K. , Webb B. , Jordan A. , Petrik J.
Published: August 2005

Background and Objectives  To reduce the risk of transfusion‐transmissible viruses entering the blood supply, the nucleic acid amplification testing (NAT) was implemented to screen Scottish and Northern Irish blood donations in minipools. After 5 years of NAT for hepatitis C virus (HCV) and 2 years for human immunodeficiency virus‐1 (HIV‐1), the yield of serologically negative, nucleic acid positive ‘window donations’ and cost‐benefit of NAT is under review.

Materials and Methods  When the Scottish National Blood Transfusion Service (SNBTS) implemented NAT in 1999, a fully automated ‘black box’ system was not available. Therefore, an ‘in‐house’ assimilated NAT assay was developed, validated and implemented. The system is flexible and allows testing for additional viral markers to be introduced with relative ease.

Results  The HCV and HIV NAT assays have 95% detection levels of 7·25 IU/ml and 39·8 IU/ml, respectively, as determined by probit analysis. One HCV (1 in 1·9 million) and one HIV (1 in 0·77 million) window donation have been detected in 5 and 2 years, respectively, of NAT.

Conclusion  The SNBTS NAT assays are robust and have performed consistently over the last 5 years. The design of the in‐house system allowed HIV NAT to be added in 2003 at a relatively small additional cost per sample, although for both assays, the royalty fee far exceeds the cost of the test itself. Clearly NAT has a benefit in improving the safety of the blood supply although the risks of transfusion‐transmitted viral infections, as reported in the Serious Hazards of Transfusion (SHOT) report, are extremely low. Also, in UK the yield of HCV antibody negative, NAT positive donations is far lower than predicted although the early detection of an HIV window period donation and the increase of HIV in the blood donor and general populations may provide a stronger case for HIV NAT.

Summary sentence  The yield of HCV and HIV NAT in UK is significantly less than that anticipated from statistical models.
Kinetic studies on Korean serum cis‐AB enzymes reveal diminished A and B transferase activities
Authors: Yazer M. H. , Zheng R. B. , Seto N. O. L. , Rose N. L. , Cho D. , Palcic M. M.
Published: August 2005

Background  cis‐AB enzymes are rare glycosyltransferases that synthesize both blood group A and B antigens. We have identified a large cohort of Korean cis‐AB blood donors and studied the N‐acetylgalactosaminyltransferase (glycosyltransferase A, GTA) and galactosyltransferase (glycosyltransferase B, GTB) activity of their cis‐AB serum enzymes.

Materials and methods  The cis‐AB01 allele was identified by PCR–RFLP (polymerase chain reaction–restriction fragment length polymorphism) in 60 donors collected at the Gwangju‐Chonnam Red Cross Blood Center. Enzyme assays of this cis‐AB enzyme were performed on available serum samples from 16 donors with the cis‐AB01/O genotype and three with the cis‐AB01/A genotype.

Results  In cis‐AB donors with an O allele, both the GTA and GTB activity of the cis‐AB enzyme were markedly reduced compared to normal A and B controls (29% and 27%, respectively). This is consistent with the behaviour predicted from kinetic studies of a recombinant model of the corresponding AAAB enzyme.

Conclusion  Although variable, cis‐AB enzymes feature reduced GTA and GTB activities.

Summary  Cis‐AB enzymes feature variable but reduced GTA and GTB activities with relatively weaker GTB activity, consistent with the weak agglutination present on forward typing with anti‐B.
Bull Spermatozoa under Mercury Stress
Authors: Arabi M
Published: August 2005

Contents

Defective sperm function is the most prevalent cause of male infertility and is difficult to treat. This study was designed to evaluate the effect of mercuric chloride (HgCl2) at 50–300 μmol/l concentration range, in vitro, on the sperm membrane and DNA integrity, viability, reduced glutathione (GSH) content and acrosomal status of the bull spermatozoa. The samples were processed for sperm analyses using semen‐diluting fluid [phosphate‐buffered saline (PBS), pH 7.2]. I recorded a meaningful increase in the lipid peroxidation (LPO) rate and a drastic fall in the spermatocrit values under mercury additions, dominantly at 300 μm mercury concentration, indicating a deleterious effect of mercury on the sperm membrane intactness. There was also a strong negative correlation between LPO rate and percentage of viable spermatozoa (r = −0.9, p < 0.001). GSH content was significantly impaired. Data obtained from Comet assay [single‐cell gel electrophoresis (SCGE)] technique revealed that mercury is capable of inducing DNA breaks in the sperm nuclei. Interestingly, 90% of DNA breaks were double‐stranded. The correlation between LPO rate and percentage of DNA breaks was found to be 0.9 (p < 0.001). Results of the gelatin test indicate that mercury is capable of altering the integrity of acrosomal membranes, showing an abnormal acrosome reaction. In this regard, a strong correlation was found between LPO rate and percentage of halos (r = −0.9, p < 0.001). In conclusion, mercury proved to be a potential oxidant in the category of ‘environmental factors’ to bull spermatozoa. Hence, considering the widespread use of mercury and its compounds, these metals should be regarded with more concern.
Effect of the Type and Number of Prostaglandin Treatments on Corpus Luteum, the Largest Follicle and Progesterone Concentration in Dairy Cows
Authors: Répási A , Beckers JF , Sulon J , Karen A , Reiczigel J , Szenci O
Published: August 2005

Contents

Lactating dairy cows (n = 72) with a mature corpus luteum (CL) (diameter of ≥17 mm) determined by ultrasonography and having a follicle with a diameter of ≥10 mm were randomly assigned to four groups. Cows were treated with cloprostenol i.m. once or twice, or with dinoprost i.m. once or twice with an 8‐h interval. The ovaries of each cow were scanned daily by transrectal ultrasonography to measure the changes in the areas of CL and the largest follicle and to determine the occurrence of ovulation. Oestrus was verified twice daily. In addition, blood sample was withdrawn from each cow daily for measuring progesterone (P4) concentrations. Significant decreases in the percentage changes relative to areas of CL and P4 concentrations or increases in the percentage changes in the area of the largest follicle on day 0 were detected in each group during the experiment. However, the type of the drug and the number of the treatments had no significant effect on those parameters. Cows ovulated with or without showing oestrus (group A) and cows exhibiting no oestrus and ovulation (group B) were also evaluated. In contrast to the mean area of the CL and the mean concentration of P4 on day 0, the mean area of the largest follicles between the two groups on day 0 differed significantly. Significant decreases in the percentage changes relative to the area of the CL and P4 concentration or increases in the percentage changes relative to the area of the largest follicle during the experiment were detected in both groups; however, there were no group differences. Treatment of dairy cows with two injections of prostaglandins (cloprostenol or dinoprost) at an 8‐h interval resulted in more cows being observed in oestrus within 5 days after treatment and having significantly higher pregnancy rate than those treated with a single prostaglandin injection.
Autopsy case of pure akinesia showing pallido‐nigro‐luysian atrophy
Authors: Konishi Yoshihiro , Shirabe Teruo , Katayama Sadao , Funakawa Itaru , Terao Akira
Published: August 2005

A 60‐year‐old man developed levodopa‐resistant pure akinesia. The patient gradually became more akinetic without accompanying gaze palsies, nuchal dystonia, or other parkinsonian features such as rigidity or tremor. At the age of 71, he died of bronchopneumonia. Neuropathologically, bilateral marked neuronal loss and gliosis were restrictedly observed in the globus pallidus, substantia nigra and corpus luysii, whereas mild gliosis without neuronal loss was found in the brain stem. With Gallyas‐Braak silver stain, numerous argyrophilic fibrous structures partly surrounding glial nuclei were observed in the three major affected regions. With Bodian stain, however, they were rarely recognized. The structures were partly positive for tau protein. Rare neurofibrillary tangles were found in the three areas and brain stem. They were relatively more numerous but still sparse in the hippocampus and the parahippocampus. The present case was diagnosed as having pallido‐nigro‐luysian atrophy based on two characteristic findings: (i) the distribution of lesions showing neuronal loss with gliosis; and (ii) significant presence of tau‐positive argyrophilic fibrous structures related to glia but with the absence of neurofibrillary tangles in the major affected regions and the brain stem. As our present case uniquely showed pure akinesia for the whole clinical course, it is noteworthy to report it here with a full neuropathological evaluation. In addition, a moderate number of diffuse plaques positive for β‐amyloid were distributed in the thalamus.
Autopsy case of acute encephalopathy linked to familial hemiplegic migraine with cerebellar atrophy and mental retardation
Authors: Takahashi Tatsuya , Arai Nobutaka , Shimamura Megumi , Suzuki Yume , Yamashita Sumimasa , Iwamoto Hiroko , Inayama Yoshiaki , Kameda Youichi , Kuroiwa Yoshiyuki
Published: August 2005

A 19‐year‐old female patient, who had exhibited esotropia, mild cerebellar ataxia, mild mental retardation, and cerebellar atrophy on magnetic resonance images at the age of 15, developed signs of acute encephalopathy, and thereafter died of disseminated intravascular coagulation on the day of her admission. Both her mother and sister suffered from attacks of hemiplegic migraine, mild mental retardation, and cerebellar ataxia. Neuropathological examinations revealed acute changes in the widespread cerebral cortex, chronic degenerative changes in the anterior lobe of the cerebellar vermis, axonal spheroids in the Goll's nucleus, pseudo‐calcinosis in the globus pallidus, and glial bundles in the cranial nerves. The most fascinating features were changes of Purkinje cells, such as cactuses (asteroid bodies, dendritic expansions), somatic sprouts, and torpedoes. These changes may be characteristic of familial hemiplegic migraine with cerebellar atrophy, as well as the other metabolic diseases, such as Menkes’ kinky hair disease, infantile (Tay–Sachs type) amaurotic idiocy, organic mercury intoxication, and mitochondrial encephalopathy, of which cases often exhibit such pathological changes of Purkinje cells. Therefore, familial hemiplegic migraine may share some metabolic abnormalities with the diseases mentioned above.