bluuebunny/crossref_metadata_2025 huggingface dataset:
Isoenzyme patterns of Blastocystis hominis patient isolates derived from symptomatic and healthy carriers
Authors: Gericke Anne‐Sophie , Burchard Gerd‐Dieter , Knobloch Jürgen , Walderich Brigitte
Published: November 2003

Isolates of Blastocystis hominis derived from patients with intestinal symptoms and from healthy carriers were cultured in vitro and isoenzyme patterns of hexokinase (E.C. 2.7.1.1), phosphoglucomutase (E.C. 2.7.5.1) and glucose phosphate isomerase (E.C. 5.3.1.9) were investigated to find evidence for pathogenic and non‐pathogenic subspecies of B. hominis. For this purpose we examined 2000 patients of the out‐patient department of the Institute for Tropical Medicine in Tübingen. From these, we obtained 232 B. hominis patient isolates; 119 isolates could be tested for the three isoenzymes. Blastocystis hominis possesses 5 patterns for hexokinase, 11 for phosphoglucomutase and 35 for glucose phosphate isomerase, showing that B. hominis is highly polymorphic. However, there was no correlation between isoenzyme patterns and disease of patients.
The hospital as an enterprise: management strategies
Authors: Stefanini Angelo
Published: November 2003

After introducing the context in which health systems exist in developing as well as in industrialized countries, the author describes the role that hospitals are expected to play as ‘enterprises’. The rationale behind such a role is explored and the experience so far available discussed. Evidence from several developing countries suggests caution and tends to discourage radical and indiscriminate introduction of market‐oriented reforms, which international donors seem to be spearheading in Africa's health systems. It is likely, however, that a limited, gradual introduction of selected elements of these reforms (such as some aspects of contracting out, managerial decentralization, the split between purchasing and providing functions) may find its place and eventually have a positive impact on the performance of the health systems. But the ‘mystique of markets’ must not be allowed to deprive public hospitals of the sense of social utility and of the caring ethos which are essential requisites of a meaningful and effective public health system.
Cultivation of bloodstream forms of Trypanosoma brucei and T. evansi in a serum‐free medium
Authors: Hirumi H. , Martin S. , Hirumi K. , Inoue N. , Kanbara H. , Saito A. , Suzuki N.
Published: November 2003

Bloodstream forms of Trypanosoma brucei and T. evansi have been cultivated in an axenic culture system by using Iscove's modified DMEM‐based HMI‐9 medium supplemented with bathocuproinedisulphonic acid, l‐cysteine, hypoxanthine, 2‐mercaptoethanol, pyruvate, thymidine and 10% fetal bovine or adult horse serum. We developed a serum‐free medium (HMI‐244) in which serum in HMI‐9 was replaced by fatty acid‐free bovine serum albumin, bovine α2‐macroglobulin, bovine β‐lipoprotein, d‐biotin, retinol, β‐alanine, l‐anserine nitrate salt, l‐ornithine hydrochloride, O‐phosphorylethanolamine, sarcosine, taurine, adenosine‐ 5′‐triphosphate, 2′‐deoxycytidine‐5′‐monophosphate, 2′‐deoxyguanosine‐5′‐monophosphate, and 5‐methyltetrahydrofolic acid. Maximum cell densities and population doubling times were 2.6×106 cells/ml; 10.6 hours and 2.2×106 cells/ml; 10.9 hours for T. brucei and T. evansi, respectively. Bloodstream forms continued to proliferate in the serum‐free cultures for more than 90 days and the trypomastigotes retained their morphological characteristics and infectivity to mice. If validated, this serum‐free medium may help reduce future interlaboratory variability in biochemical, immunological, molecular biological and drug sensitivity studies on these parasites.
A health insurance scheme for hospital care in Bwamanda district, Zaire: lessons and questions after 10 years of functioning
Authors: Criel Bart , Kegels Guy
Published: November 2003

A voluntary insurance scheme for hospital care was launched in 1986 in the Bwamanda district in North West Zaire. The paper briefly reviews the rationale, design and implementation of the scheme and discusses its results and performance over time. The scheme succeeded in generating stable revenue for the hospital in a context where government intervention was virtually absent and external subsidies were most uncertain. Hospital data indicate that hospital services were used by a significantly higher proportion of insured patients than uninsured people. The features of the environment in which the insurance scheme thrived are discussed and the conditions that facilitated its development reviewed. These conditions comprise organizational‐managerial, economic‐financial, social and political factors. The Bwamanda case study illustrates the feasibility of health insurance – at least for hospital‐based inpatient care – at rural district level in sub‐Saharan Africa, but also exemplifies the managerial and social complexity of such financing mechanisms.
Cytokine levels during mild and cerebral falciparum malaria in children living in a mesoendemic area
Authors: Baptista João Luís , Vanham Guido , Wéry Marc , Van Marck Eric
Published: November 2003

Cell‐mediated immunity and cytokines are probably involved in the pathogenesis of malaria. To investigate the role and the activity of different immune cells, we measured levels of tumour necrosis factor‐(TNF‐α), gamma interferon (IFN‐γ) and several interleukins (IL‐2, IL‐4, IL‐6 and IL‐10) in children with mild (MM) and cerebral (CM) Plasmodium falciparum malaria and compared them with those of healthy children from Guadalupe – Lobata District, St. Tomé Island, where malaria is mesoendemic. Both groups of patients had significantly higher levels of IL‐6, IL‐10 and TNF‐α than controls. For IL‐2, IL‐4 and IFN‐γ we found no difference between the groups. However, 24 h after admission the levels of IL‐10 and IL‐6 were significantly higher in CM than in MM patients, although 7 days after treatment they returned to normal levels, similar to those found in control children. Therefore, TNF‐α IL‐6 and IL‐10 increase during Plasmodium falciparum attacks in all children, not only in those with cerebral malaria. This finding suggests the activation of the monocyte/macrophage system during the early stage of clinical malaria.
A population‐based coprological study of human fascioliasis in a hyperendemic area of the Bolivian Altiplano
Authors: Esteban J. G. , Flores A. , Angles R. , Strauss W. , Aguirre C. , Mas‐Coma S.
Published: November 2003

The community of Chijipata Alta, at an altitude of 3850 m, near the southern coast of Lake Titicaca in the Northern Altiplano of Bolivia, was surveyed for human fascioliasis. The global prevalence (66.7%) and intensity (eggs per gram of faeces – epg: range: 24–4440; arithmetic mean: 1001; geometric mean: 390) proved to be the highest known in the world by means of coprological techniques. These results suggest the existence of highly hyperendemic subzones among the large human fascioliasis‐endemic zone of the Bolivian Northern Altiplano. Despite the decrease in prevalence and intensity from children (75.0%, 24–4440 epg) to adults (41.7%, 144–864 epg), our findings show that in an hyperendemic zone adult subjects either maintain the parasites acquired when young or are newly infected as the consequence of inhabiting a zone of high infection risk.
Five subgroups of Blastocystis hominis isolates from symptomatic and asymptomatic patients revealed by restriction site analysis of PCR‐amplified 16S‐like rDNA
Authors: Böhm‐Gloning B. , Knobloch J. , Walderich B.
Published: November 2003

DNA polymorphism of Blastocystis hominis isolates was examined by the amplification of a gene fragment coding for the 16S‐like rRNA. Using identical primers, fragments of approximately 850 bp were amplified from 110 B. hominis isolates and fragments of 1.1kbp were amplified from 48 isolates. Digestion of the amplification products with the restriction enzymes HinfI, RsaI, and AluI revealed different profiles for each fragment length. Subgroup I and II, resulting from digestion of the smaller 850 bp fragment, have identical HinfI and AluI restriction bands, subgroup III and IV have identical RsaI fragments after digestion of the 1.1kbp DNA. Subgroup V resembles subgroup III in a few bands after the RsaI and AluI restriction, respectively. Ninety (54%) of the isolates studied were assigned to subgroup I, 20 (12%) to subgroup II, 35 (21%) to subgroup III, 12 (7%) to subgroup IV, and 1 (1%) to subgroup V. Five (3%) of the examined people were coinfected with B. hominis of subgroup I and III, 3 (2%) carried B. hominis of subgroup I and II. These results show that there are 5 B. hominis subgroups none of which was found to be significantly correlated with the reported disease.
Subclass‐specific serological reactivity and IgG4‐specific antigen recognition in human echinococcosis
Authors: Dreweck C. M. , Lüder C. G. K. , Soboslay P. T. , Kern P.
Published: November 2003

Immunglobulin (Ig) subclass‐specific antibody responses and isotype‐specific recognition of E. multilocularis (Em) and E. granulosus (Eg) antigens (Ag) were evaluated in both alveolar echinococcosis (AE) and cystic echinococcosis (CE). AE patients were divided into 3 groups by clinical and therapeutic criteria according to their actual state of infection, i.e. elimination of parasite, and regression or progression of disease. CE patients were either before or after surgery, or in continuous chemotherapy due to parasite persistence. Total IgE was highly elevated in progressive AE cases (7/11), but not in the cases with eliminated infection or regression. In AE patients with active disease, EmAg‐specific IgE, total IgG, IgG1, IgG2 and IgG4 were particularly high. Similarly, in 9 of 30 CE patients, total IgE was raised above reference values, indicating progressive disease. CE patients' sera antibody cross‐reacted with crude EmAg, and detectable Ig levels of the same isotype were also measured by ELISA. In both AE and CE, parasite‐specific antigen recognition was dominated by IgG1 and IgG4. In AE patients with progressive disease, IgG4 distinctively recognized low molecular weight EmAg of Mr 26 kD, 18 kD, 16 kD and 12 kD. As prominent IgG4 and IgE responses develop with chronic helminth infections only, these serological parameters may indicate successful parasite infestation and severe outcome of disease. In summary, analyses of immunoglobulin isotype responses in AE patients by ELISA in combination with immunoblotting are a useful approach for post‐treatment follow‐up of patients at risk of developing recrudescent disease.
Granulomatous dermatitis caused by Mycobacterium genavense in two psittacine birds
Authors: FERRER LUIS , RAMIS ANTONIO , FERNÁNDEZ JESÚS , MAJÓ NATÀLIA
Published: November 2003

Two cases of cutaneous mycobacteriosis in psittacine birds showing featherless, non‐painful, non‐pruritic nodules are described. Histopathological studies of skin biopsies from both cases demonstrated the presence of a diffuse granulomatous dermatitis with acid‐ fast organism s inside macrophages, which led to the diagnosis of cutaneous mycobacteriosis. In one case, generalization of the process to internal organs (intestinal and hepatic serosae) was observed. Mycobacterial organisms could not be cultured using conventional isolation media (Coletsos and Löwenstein–Jensen), but polymerase chain reaction (PCR) technique performed on pathological samples from both birds revealed the presence of Mycobacterium genavense. It is thus proposed that cutaneous mycobacteriosis infections, in particular those caused by M.genavense, should be included in the differential diagnosis of skin nodular processes in psittacine birds. The usefulness of PCR techniques for aetiological diagnosis of mycobacterial infections is emphasized.
The microclimate of the canine coat: the effects of heating on coat and skin temperature and relative humidity
Authors: Chesney C. J.
Published: November 2003

The coat of seven ‘Landseer’ Newfoundland dogs was irradiated using an infra‐red source for 25 min. In each dog, at a site of each colour (black and white), skin and coat temperatures were monitored, and coat air humidity measured with a specially designed instrument. Almost no differences were noted at sites with differing coat colour. Skin temperature rose from 35 °C to a plateau at 39 °C, whilst coat temperature rose from 30 °C to 41 °C. Relative humidity of coat air initially rose, then fell significantly (P < 0.001). The absolute humidity initially almost doubled (P < 0.001), but then fell, although remaining significantly higher than that of ambient air. It was concluded that an initial burst of sweating was followed by lower but continuing secretion. This was not, however, of great importance in cooling. In a separate study the skin temperature of black coated dogs exposed to bright sunshine was explored. The mean temperature was almost identical to that of the plateau skin temperature noted in Newfoundland dogs.
Malassezia dermatitis in the dog: a retrospective histopathological and immunopathological study of 86 cases (1990–95)
Authors: MAULDIN ELIZABETH A. , SCOTT DANNY W. , MILLER WILLIAM H. , SMITH CHRISTINA A.
Published: November 2003

A retrospective histopathological and immunopathological study was conducted on 86 dogs with Malassezia dermatitis. West Highland White terriers, English Setters, Shih Tzus, Basset Hounds, American Cocker Spaniels, spayed females, and castrated males were found to be at increased risk. The histopathological reaction pattern of lymphocytic superficial perivascular to interstitial dermatitis with parakeratotic hyperkeratosis, irregular epidermal hyperplasia, diffuse intercellular oedema and lymphocytic exocytosis was found to be consistent with a diagnosis of Malassezia dermatitis whether yeast were histologically visible (73.3% of the cases) or not (26.7%). Immunopathological studies revealed that 60– > 90% of the inflammatory cells within the epidermis, and 25–75% of those within the dermis were CD3+T lymphocytes, and that the only immunoglobulin‐positive cells were dermal plasma cells.
Skin and mucosal populations of Malassezia pachydermatis in healthy and seborrhoeic Basset Hounds
Authors: Bond R. , Lloyd D.H.
Published: November 2003

Quantitative and semiquantitative cultural techniques were used to study skin and mucosal carriage of Malassezia pachydermatis in 20 healthy mixed‐breed dogs, 13 healthy Basset Hounds and 33 seborrhoeic Basset Hounds. The frequencies of isolation and population sizes from the axilla, nose, mouth and vulva were significantly greater (P < 0.01) in both groups of Basset Hounds when compared with the healthy mixed‐breed dogs. Population sizes in the seborrhoeic Basset Hounds exceeded those of the healthy bassets at the nose (P < 0.05), vulva (P < 0.01) and axilla (P < 0.001). However, the frequencies of isolation and population sizes of the yeast from the anus were comparable in the three groups of dogs. The relatively high skin and mucosal populations in healthy Basset Hounds may explain, in part, the predisposition to ‘Malassezia dermatitis’ in this breed. However, the factors which enable the yeast to establish such high populations in Basset Hounds remain unclear.
Response of an acetolactate synthase (ALS)‐resistant biotype of Amaranthus rudis to selected ALS‐inhibiting and alternative herbicides
Authors: SPRAGUE C. L. , STOLLFR E. W. , WAX L. M.
Published: November 2003

An Amaranthus rudis Saner (common water‐hemp) biotype from a field treated for two consecutive years with a mixture of chlorimuron and metribuzin was tested in greenhouse and laboratory studies to assess resistance and cross‐resistance to four acetolactale synthase (ALS)‐inhibiting herbicides. The biotype demonstrated >1920‐fold resistance at the whole plant level, and >850‐fold resistance at the ALS enzyme to chlorimuron, compared with a susceptible biotype. This chlorimuron‐resistant biotype was also cross‐resistant to primisuifu‐ron, haiosulfuron and imazethapyr. In greenhouse studies, atrazine alone or in combination with ALS‐inhibiting herbicides provided excellent control of the resistant biotype of A. rudis. Combinations of dicamba and ALS‐inhibitors also provided adequate control. Additionally, premixtures of flumetsulam and metolachlor and of dicamba and atrazine furnished excellent control of this chlorimuron‐resistant A. rudis biotype.
Germination behaviour of two morphologically different types of seed of Cassia tora at different temperatures
Authors: BHATTACHARYA A. , SAHA P. K.
Published: November 2003

The effect of temperature on the germination response of two categories of morphologically different seeds of Cassia tora L. was investigated. Germination response of dormant seeds (category A with closed ultraastructures) was not affected by temperature, as 96% of the seeds germinated at all temperatures on scarification. On the other hand, non‐dormant seeds (category B with open ultrastructures) were temperature sensitive and germinated best at 32°C. However, storage affected the germination responses of both types. While the category A seeds lost moisture gradually with storage time and retained viability for a longer period, i.e. 2 years, the moisture loss in category B seeds was noted only after 2 years and the seed viability was retained for only 1 year. The production of two morphologically and physiologically distinct types of seed simultaneously by C. tora is probably an ecological adaptation for progressive emergence of the species over long periods in the field.
Dose response of weeds to methyl iodide and methyl bromide
Authors: ZHANG W. M. , McGIFFEN M. E. , BECKER J. O. , OHR H. D. , SIMS J. J. , KALLENBACH R. L.
Published: November 2003

Labonuory bioassay and field experiments were conducted to characterize the dose response of weeds to methyl iodide and methyl bromide as soil fumigants. The patterns in potency of both fumigants and in sensitivity of diffcretit weed species to the fumigants were distinguished with the use of logistic dose‐response models. Similar to its response to methyl hromide fumigation. Amaranthus retrofleus L. was the most sensitive to methyl iodide fumigation. Cyperus rotundtis L. was the least sensitive to methyl iodide fumigation, whereas Portuloca oleracea L. was the least sensitive to methyl hromide. Lolium multiflorum Lam. Abutilon theophrasti Medik.Chenopodium album L. P. ateracea. Brassica kaber (D.C.) L.C. Wheeler and Cyperus escuden‐tus L. were similar in sensitivity to methyl iodide. Methyl iodide was as potent as methyl bromide for A. retroflexus but more potent than methyl bromide for L. multiflorum, A. theophrasti. C. album. P. oleracea. B. kaber, C. esculentus and C. rotundus. The dose response for weeds in the field was similar to that obtained in laboratory bioassays. Under fieid conditions. 280 kg ha‐1 methyl iodide killed all species tested except Solanum nigrum L Methyl iodide appears to be a suitable replacement for meihyl bromide because it can be used in situations simitar to methyl bromide fumigation, has superior efficacy against a broad spectrum of pests and has a low potential for degrading the earth's ozone lavers.
A model of the cost of delay in spraying weeds in cereals
Authors: BLACK I. D. , DYSON C.B.
Published: November 2003

Using a previous model as a base, data derived in field trials in wheat (Triticum aestivuort L.) and bailey (Hordeum vulgare L.) were assessed to estimate the yield penalty, and resulting reduction in economic benefit, from progressive spraying of herbicides up to late crop tillering. Compared with early post‐emergence application, yield penalties from delayed spraying began at early tillering. For median values of weed‐free yield and weed density in the data set. the loss in potential yield increase when spraying at late tillering compared with early post‐emergence was 71%, Crops with higher weed‐free yield potential and with greater initial weed density showed a proportionately (as well as absolutely) larger yield penalty from a moderate delay in spraying, indicating an earlier and more intense onset of competition with weeds.
Growth and regenerative capacity of plants of Cirsium arvense
Authors: GUSTAVSSON ANN‐MARIE DOCK
Published: November 2003

The undisturbed growth and regenerative capacity of the root system of Cirsium arvense (L.) Scop, were studied in one glasshouse and one field experiment. In the glasshouse experiment, two lengths of root fragment from the same clone, previously grown at two nitrogen levels, were planted in boxes. Plants that developed from these fragments were inspected on eight occasions to assess their undisturbed development and growth, above and below ground, and their regenerative capacity after disturbance by burial at different points of development. The initial nitrogen content of the roots had no effect on growth measurements, but longer root fragments produced greater masses of primary shoots, new roots and shoots from the new roots than did shorter ones. After disturbance at early stages (4‐7 leaves with a minimum length of 5 cm), plants originating from shorter root fragments did not regrow at all. Plants originating from longer root fragments exhibited a minimum regenerative capacity when disturbed at the eight‐leaf stage. In the field experiment, root fragments of length 12 cm. from the same clone, were planted in October 1991 at 5 cm or 20 cm depth. Plants that developed from these fragments were studied on 12 occasions, from April to July 1992. Plants emerging from a depth of 20 cm reached a point of positive net assimilation 28 days (380 day degrees) later than those developing from a depth of 5 em. The plants exhibited a minimum regenerative capacity when their primary shoots had. on average. 10 (plants from 5‐cm root fragments) or seven (plants from 20‐cm root fragments) expanded leaves.
Horizontal movement of seeds following tine and plough cultivation: implicationsfor spatial dynamics of weed infestations
Authors: REW L. J. , CUSSANS G. W.
Published: November 2003

Horizontal movement of seeds by different cultivators was investigated in three experiments. Seeds of barley (Hordeum vulgare L.), field bean (Vicia faba L.) and oilseed rape (Brassica napus L.) were positioned on the soil surface or buried 0.1 m deep prior to cultivation, Seed movement was assessed by counting germinated seedlings. The type of tine implement used significantly affected mean seed movement, with surface‐sown seeds being moved significantly further than buried seeds. Primary cultivation with flexi‐ or spring‐tine machines moved seeds further than straight‐tine or power harrow implements. However, there was no overall difference between plough and flexi‐tine mean seed movement. A single pass with a tine attached to a seed drili moved seeds less than the two passes with tine plus seed drill, but the first pass had the greatest effect. Small oilseed rape seeds moved significantly further than larger barley or field bean seeds. More than 84% of seeds moved ≤ 1m from the source: no seeds were observed>4.8m in a forward direction or >0.2m backwards from the source, In a fourth experiment, plots containing seeds of Sinapis arvensis L. were cultivated in alternate directions in successive years, The results broadly validated the findings of the previous experiments Quantifying the horizontal movement of weed seeds is important to the understanding of the spatial dynamics of weed patches and thus in the prediction of future weed distributions.
Germination behaviour of dormant oilseed rape seeds in relation to temperature
Authors: PEKRUN C. , LUTMAN P. J. W. , BAEUMER K.
Published: November 2003

Seeds of oilseed rape (Brassica napus L., cv. Liglandor) were induced into secondary dormancy by imbibing them for 2‐4 weeks in an osmotic solution at ‐1500 kPa in darkness. Treated seeds were then transferred into pots or Petri dishes, where they were given adequate water for germinatton. These pots and Petri dishes were exposed to various temperature regimes during the course of the following 6–14 months. During that time, seed samples were taken at 2‐ to 6‐ weekly intervals to monitor survival of seeds and to study their dormancy. The ntimber of seeds that had not germinated in pots or Petri dishes remained constant in the majority of treatments. but it declined when seeds were exposed to pronounced daily temperature alternations. This decline was due to germination and not to death of seeds from other causes. Germination tests on subsamples of seeds showed that they were light sensitive, more seeds germinating in the light than in the dark. The seeds’germination behaviour differed between samphng dates. This variation in germination behaviour appeared not to display the classical dormancy:non‐dormancy cycle. Part of the variation seemed to be due to the changes in temperature that the seeds experienced when taken from pots and set up in the germination tests. Seeds exposed to a constant temperature for 1 year appeared lo become more dormant with time. The relevance of this work in relation to the persistence of volunteer oilseed rape is discussed.
Effects of nitrogen supply on competition between wheat and three annual weed species
Authors: IQBAL J. , WRIGHT D.
Published: November 2003

Three pot experiments have investigated the effects of nitrogen (N) supply on interspecific competition between three weed species (Phalaris minor Retz., Chenopodium album L, and Sinapis arvensis L.) and spring wheat (Triticum aestivum L.) cv, Alexandria. The treatments tested included monocultures of each species and a mix‐ture containing them in equal proportions that were combined factorially with two levels of N supply (20, 120 kg N ha‐1). Low N supply decreased net photosynthetic rate (Pn), leaf N percentage, plant dry weight and N uptake of both wheat and weed species and gram dry weight of wheat. The effects of low N on Pn and dry weight of weeds were greater than the effects on wheat. In most cases the decrease in Pn at low N was due to non‐stomatal factors. The relative competitive abilities of wheat and weeds were influenced by N supply. At high N, S.arvensis was more competitive than wheat, whereas P. minor was less competitive than wheat. C. album was more competitive than wheat at both N levels. The rank order of competitive ability of the weed species was C. album > P. minor > S. arvensis. The effects of interspecific competition on Pn were smaller than the effects of N supply and were not associated with corresponding effects on leaf N percentage and plant dry weight of both wheat and weed species and grain dry weight of wheat.
Use of the extended BBCH scale—general for the descriptions of the growth stages of mono; and dicotyledonous weed species
Authors: HESS M. , BARRALIS G. , BLEIHOLDER H. , BUHR L. , EGGERS TH. , HACK H. , STAUSS R.
Published: November 2003

The extended BBCH scale is a system for a uniform coding of phenologicaliy similar growth stages of all mono‐ and dicotyledonous plant species, based on the well known cereal code of Zadoks et al. (1974). The BBCH key is it decimal system, with 10 principal growth stages and up to 10 secondary ones, starting with seed germination, sprouting of perennials, progressing through leaf production and extension growth to flowering and senescence. Therefore, it can also be a suitable tool to define the growth stages of different weed species. To encourage further use of the BBCH scale in weed research, definitions of the codes have been more closely adapted to weeds. Possible problems are discussed and guidelines for correct use are given.
Serological characteristics of peanut allergy in children
Authors: Bernard H. , Paty E. , Mondoulet L. , Burks A. W. , Bannon G. A. , Wal J. M. , Scheinmann P.
Published: November 2003

Background:  Food challenge is considered an excellent clinical tool for the diagnosis of specific food allergy. However in the case of peanut allergy it may be difficult to perform because of the severity of the reactions. The quantitation of a specific immunoglobulin E (IgE) response to different peanut allergens could also contribute to the improvement of the diagnosis. We characterized the IgE response to a whole peanut protein extract and to Ara h 1 and Ara h 2 in different groups of patients classified according to the severity of their allergic reactions.

Methods:  Specific serum IgE were analyzed in 96 children by enzyme‐linked immunosorbent assay tests using a whole protein extract or purified peanut proteins and anti‐human IgE monoclonal antibodies labeled with acetylcholinesterase.

Results:  A parallel was observed between levels of peanut‐specific IgE and the classification in five groups and subgroups of patients upon increasing severity of symptoms, especially within the group of highest severity. Moreover, the highest frequency of positive response and the highest levels of specific IgE were observed with whole peanut protein extract.

Conclusion:  In a retrospective evaluation of peanut allergy in children, we have shown that quantitation of peanut‐specific IgE could be used to avoid a food challenge particularly in the case of severe reactions. When compared to Ara h 1 and Ara h 2, whole peanut protein extract appeared to be the most appropriate allergen to perform the test.
Allergenicity of varieties of soybean
Authors: Codina R. , Ardusso L. , Lockey R. F. , Crisci C. , Medina I.
Published: November 2003

Background:  Soybean hulls (SHs) cause respiratory allergies. This study investigates the allergenicity of soybean varieties (SVs) by in vivo and in vitro tests.

Methods:  Ten SVs were studied: (a) five with a proved clinical relevance (SVs 1, 2, 3, 4, 5), the last four with a ‘dull’ phenotype; (b) five of undetermined relevance, three of them (SVs 6, 7, 8) with a ‘shiny’ phenotype, and two (SVs 9 and 10) with a ‘dull’ phenotype. Extracts from all 10 SVs were used to skin prick test (SPT) 21 subjects sensitized to SHs. Positive and negative sera pools prepared from sera of subjects sensitized or not to SHs, respectively, were utilized to perform in vitro experiments (specific IgE and IgG4 determinations, SDS‐PAGE/IgE‐Western blot, and IgE‐inhibition).

Results:  In this study, it was found that 52.4, 52.4, 57.1, 71.4, 80.9, 42.9, 57.1, 71.4, 52.4, and 38.1% subjects had a positive SPT with SVs 1–10, respectively (P NS). Specific IgE values to SVs 1–10 obtained with the positive pool are 28.3, 26.4, 29.9, 28.3, 26.8, 4.8, 13.4, 6.7, 24.7, and 17.5% total counts bound, respectively; and specific IgG4 values 0.851, 0.818, 0.721, 1.609, 0.789, 0.617, 0.662, 0.0, 1.127, and 0.934 OD units, respectively; the microgram of protein required to produce 50% inhibition are 2.5, 3.7, 4.5, 2.4, 5, 39.8, 25.2, 25.1, 4.5, and 8.9, respectively. A 7‐kDa band is present in all SVs except in those with a ‘shiny’ phenotype.

Conclusions:  The SVs with a ‘shiny’ phenotype contain less allergens than the other SVs studied, as determined by in vitro tests. However, SPT results with the SVs do not differ. Genetic screens should be devised to select plants with reduced, preferably absent, allergenicity, but with a high nutritional value, and this allergenicity should be studied utilizing in vivo and in vitro tests.
Future therapeutic options in food allergy
Authors: Eigenmann P. A.
Published: November 2003

Up to 5% of young children and 2% of adults suffer from food allergy. Among them many have immunoglobulin E (IgE)‐mediated food allergy, a condition with potentially fatal allergic reactions. Several studies have addressed possible definite treatment options for food allergy. Immunotherapy, by the oral route or by systemic injections shows promising preliminary results, but current interpretation of these therapeutic options are mostly handicapped by studies with insufficient scientific support, or by severe side‐effects. Currently, no studies can support pharmacotherapy. Finally, most promising results were recently published with anti‐IgE antibodies in a human trial, or various approaches in a mouse model of food allergy (chinese herbal medicine, specific modulation of the T cell response). Rapidly evolving findings might provide hope for a cure of food allergy in the near future.
Prediction of buckwheat allergy using specific IgE concentrations in children
Authors: Sohn M. H. , Lee S. Y. , Kim K.‐E.
Published: November 2003

Introduction:  Buckwheat (BW) is considered to be one of the most important food allergens, and positive skin tests are found in about 5% of Koreans. We investigated the positive and negative predictive values of BW‐specific IgE in subjects with a BW allergy in order to reduce the need for buckwheat challenge, which can be more riskier than other causes of food allergies.

Methods:  Twenty‐eight BW allergic subjects with symptoms after BW Open food challenge and 16 asymptomatic control subjects with positive skin test to BW were recruited. Serum samples from all patients were analyzed for BW‐specific IgE antibodies using the Pharmacia CAP System.

Results:  According to the receiver operator characteristic (ROC) analysis, the optimal cutoff level of BW‐specific IgE, as the definitions of serum BW‐specific IgE positive, was 1.26 kUA/l. With this selected cutoff level, the sensitivity, specificity, positive and negative predictive values were 93.10, 93.33, 79.75 and 97.96%, respectively.

Conclusion:  The use of the optimal cutoff level, 1.26 kUA/l, that simultaneously maximizes sensitivity and specificity, would be helpful for avoiding unnecessary risky challenge in children with a strong clinical history and skin test responses.
Clinical and immunological effects of low‐dose IFN‐α treatment in patients with corticosteroid‐resistant asthma
Authors: Simon H.‐U. , Seelbach H. , Ehmann R. , Schmitz M.
Published: November 2003

Background:  Interferon (IFN)‐α is a cytokine that possesses potent anti‐viral and immunoregulatory activities. We aimed to assess clinical and immunological effects of low‐dose IFN‐α in patients with severe corticosteroid‐resistant asthma with and without Churg–Strauss syndrome. There is currently no efficient pharmacological treatment available for this group of patients.

Methods:  We studied 10 patients with corticosteroid‐resistant asthma, in which 3×106 IU/day IFN‐α were administrated in addition to the prednisone dose given already before introduction of the cytokine therapy. The prednisone dose was gradually reduced dependent on the clinical situation and used as a clinical readout to evaluate the efficacy of the cytokine therapy. To distinguish between IFN‐α‐ and prednisone‐mediated immunological changes, the corticosteroid dose was kept constant for at least 2 weeks upon introduction of the cytokine therapy in seven patients. The effects of treatment on clinical and immunological parameters were measured at 2–4 weeks and 5–10 months depending on the availability of the patient.

Results:  Interferon‐α treatment rapidly improved the clinical situation as assessed by lung function parameters and required prednisone dose. Important immunological changes included: decreased leukocyte numbers, increased relative numbers of CD4+ T cells, increased differentiation of T helper (Th)1 cells, and increased expression of interleukin (IL)‐10 in peripheral blood mononuclear cells.

Conclusion:  Interferon‐α treatment was associated with dramatic improvements in the condition of patients with corticosteroid‐resistant asthma with and without Churg–Strauss syndrome. Potential mechanisms of action include the establishment of a correct Th1/Th2 balance and the induction of the anti‐inflammatory IL‐10 gene.
An unusual amide synthetase (CouL) from the coumermycin A1 biosynthetic gene cluster from Streptomyces rishiriensis DSM 40489
Authors: Schmutz Elisabeth , Steffensky Marion , Schmidt Jürgen , Porzel Andrea , Li Shu‐Ming , Heide Lutz
Published: November 2003

The aminocoumarin antibiotic coumermycin A1 produced by Streptomyces rishiriensis DSM 40489 contains two amide bonds. The biosynthetic gene cluster of coumermycin contains a putative amide synthetase gene, couL, encoding a protein of 529 amino acids. CouL was overexpressed as hexahistidine fusion protein in Escherichia coli and purified by metal affinity chromatography, resulting in a nearly homogenous protein. CouL catalysed the formation of both amide bonds of coumermycin A1, i.e. between the central 3‐methylpyrrole‐2,4‐dicarboxylic acid and two aminocoumarin moieties. Gel exclusion chromatography showed that the enzyme is active as a monomer. The activity was strictly dependent on the presence of ATP and Mn2+ or Mg2+. The apparent Km values were determined as 26 µm for the 3‐methylpyrrole‐2,4‐dicarboxylic acid and 44 µm for the aminocoumarin moiety, respectively. Several analogues of the pyrrole dicarboxylic acid were accepted as substrates. In contrast, pyridine carboxylic acids were not accepted. 3‐Dimethylallyl‐4‐hydroxybenzoic acid, the acyl component in novobiocin biosynthesis, was well accepted, despite its structural difference from the genuine acyl substrate of CouL.
Mutations in the C‐terminal domain of ALSV (Avian Leukemia and Sarcoma Viruses) integrase alter the concerted DNA integration process in vitro
Authors: Moreau Karen , Faure Claudine , Violot Sébastien , Verdier Gérard , Ronfort Corinne
Published: November 2003

Integrase (IN) is the retroviral enzyme responsible for the integration of the DNA copy of the retroviral genome into the host cell DNA. The C‐terminal domain of IN is involved in DNA binding and enzyme multimerization. We previously performed single amino acid substitutions in the C‐terminal domain of the avian leukemia and sarcoma viruses (ALSV) IN [Moreau et al. (2002). Arch. Virol.147, 1761–1778]. Here, we modelled these IN mutants and analysed their ability to mediate concerted DNA integration (in an in vitro assay) as well as to form dimers (by size exclusion chromatography and protein–protein cross‐linking). Mutations of residues located at the dimer interface (V239, L240, Y246, V257 and K266) have the greatest effects on the activity of the IN. Among them: (a) the L240A mutation resulted in a decrease of integration efficiency that was concomitant with a decrease of IN dimerization; (b) the V239A, V249A and K266A mutants preferentially mediated non‐concerted DNA integration rather than concerted DNA integration although they were found as dimers. Other mutations (V260E and Y246W/ΔC25) highlight the role of the C‐terminal domain in the general folding of the enzyme and, hence, on its activity. This study points to the important role of residues at the IN C‐terminal domain in the folding and dimerization of the enzyme as well as in the concerted DNA integration of viral DNA ends.
Oxygen tension regulates the expression of a group of procollagen hydroxylases
Authors: Hofbauer Karl‐Heinz , Gess Bernhard , Lohaus Christiane , Meyer Helmut E. , Katschinski Dörte , Kurtz Armin
Published: November 2003

In this study, we have characterized the influence of hypoxia on the expression of hydroxylases crucially involved in collagen fiber formation, such as prolyl‐4‐hydroxylases (Ph4) and procollagen lysyl‐hydroxylases (PLOD). Using the rat vascular smooth muscle cell line A7r5, we found that an hypoxic atmosphere caused a characteristic time‐dependent five‐ to 12‐fold up‐regulation of the mRNAs of the two P4h α‐subunits [αI (P4ha1) and αII (P4ha2)] and of two lysylhydroxylases (PLOD1 and PLOD2). These effects of hypoxia were mimicked by the iron‐chelator deferoxamine (100 µm) and by cobaltous chloride (100 µm). The hypoxic induction of these genes was also seen in the mouse juxtaglomerular As4.1 cell line and mouse hepatoma cell line Hepa1 but was almost absent in the mutant cell line Hepa1C4, which is defective for the hypoxia‐inducible transcription factor 1 (HIF‐1). In addition, the enzyme expression was induced by hypoxia in mouse embryonic fibroblasts but not in embryonic fibroblasts lacking the HIF‐1α subunit. These findings indicate that hypoxia stimulates the gene expression of a cluster of hydroxylases that are indispensible for collagen fiber formation. Strong indirect evidence, moreover, suggests that the expression of these enzymes during hypoxia is coordinated by HIF‐1.
The distinct nucleotide binding states of the transporter associated with antigen processing (TAP) are regulated by the nonhomologous C‐terminal tails of TAP1 and TAP2
Authors: Bouabe Hicham , Knittler Michael R.
Published: November 2003

The transporter associated with antigen processing (TAP) delivers peptides into the lumen of the endoplasmic reticulum for binding onto major histocompatibility complex class I molecules. TAP comprises two polypeptides, TAP1 and TAP2, each with an N‐terminal transmembrane domain and a C‐terminal cytosolic nucleotide binding domain (NBD). The two NBDs have distinct intrinsic nucleotide binding properties. In the resting state of TAP, the NBD1 has a much higher binding activity for ATP than the NBD2, while the binding of ADP to the two NBDs is equivalent. To attribute the different nucleotide binding behaviour of NBD1 and NBD2 to specific sequences, we generated chimeric TAP1 and TAP2 polypeptides in which either the nonhomologous C‐terminal tails downstream of the Walker B motif, or the core NBDs which are enclosed by the conserved Walker A and B motifs, were reciprocally exchanged. Our biochemical and functional studies on the different TAP chimeras show that the distinct nucleotide binding behaviour of TAP1 and TAP2 is controlled by the nonhomologous C‐terminal tails of the two TAP chains. In addition, our data suggest that the C‐terminal tail of TAP2 is required for a functional transporter by regulating ATP binding. Further experiments indicate that ATP binding to NBD2 is important because it prevents simultaneous uptake of ATP by TAP1. We propose that the C‐terminal tails of TAP1 and TAP2 play a crucial regulatory role in the coordination of nucleotide binding and ATP hydrolysis by TAP.
Prevention of thermal inactivation and aggregation of lysozyme by polyamines
Authors: Kudou Motonori , Shiraki Kentaro , Fujiwara Shinsuke , Imanaka Tadayuki , Takagi Masahiro
Published: November 2003

Proteins tend to form inactive aggregates at high temperatures. We show that polyamines, which have a relatively simple structure as oligoamids, effectively prevent thermal inactivation and aggregation of hen egg lysozyme. In the presence of additives, including arginine and guanidine (100 mm), more than 30% of 0.2 mg·mL−1 lysozyme in sodium phosphate buffer (pH 6.5) formed insoluble aggregates by heat treatment (98 °C for 30 min). However, in the presence of 50 mm spermine or spermidine, no aggregates were observed after the same heat treatment. The residual activity of lysozyme after this heat treatment was very low (< 5%), even in the presence of 100 mm arginine and guanidine, while it was maintained at ≈ 50% in the presence of 100 mm spermine and spermidine. These results imply that polyamines are new candidates as molecular additives for preventing the thermal aggregation and inactivation of heat‐labile proteins.
A differential scanning calorimetry study of tetracycline repressor
Authors: Kędracka‐Krok Sylwia , Wasylewski Zygmunt
Published: November 2003

Tetracycline repressor (TetR), which constitutes the most common mechanism of bacterial resistance to an antibiotic, is a homodimeric protein composed of two identical subunits, each of which contains a domain possessing a helix–turn–helix motif and a domain responsible for binding tetracycline. Binding of tetracycline in the protein pocket is accompanied by conformational changes in TetR, which abolish the specific interaction between the protein and DNA. Differential scanning calorimetry (DSC) and CD measurements, performed at pH 8.0, were used to observe the thermal denaturation of TetR in the absence and presence of tetracycline. The DSC results show that, in the absence of tetracycline, the thermally induced transitions of TetR can be described as an irreversible process, strongly dependent on scan rate and indicating that the protein denaturation is under kinetic control described by the simple kinetic scheme: , where k is a first‐order kinetic constant, N is the native state, and D is the denatured state. On the other hand, analysis of the scan rate effect on the transitions of TetR in the presence of tetracycline shows that thermal unfolding of the protein can be described by the two‐state model: . In the proposed model, TetR in the presence of tetracycline undergoes co‐operative unfolding, characterized by an enthalpy change (ΔHcal = 1067 kJ·mol−1) and an entropy change (ΔS = 3.1 kJ·mol−1).
Mechanisms of motor vehicle accidents that predict major injury
Authors: Palanca Sylvia , Taylor David McD , Bailey Michael , Cameron Peter A
Published: November 2003

Objective: To assess whether prehospital triage guidelines, based on mechanistic criteria alone, accurately identify victims of motor vehicle accidents (MVA) with major injury.

Methods: Retrospective analysis of the Royal Melbourne Hospital trauma database. Mechanisms analysed were those outlined by the American College of Surgeons Committee on Trauma and Advanced Trauma Life Support/Early Management of Severe Trauma prehospital triage guidelines.

Results: There were 621 MVA analysed, 253 with major injury (40.7%). Multivariate logistic regression indicated prolonged extrication time (P < 0.0001), cabin intrusion (P = 0.047), high speed (P = 0.003) and ejection from vehicle (P = 0.04) were statistically associated with major injury. Vehicle rollover and fatality in the same vehicle were not statistically associated with major injury.

Conclusions: These data suggest that existing guidelines for the prehospital triage of MVA victims, based on mechanistic criteria alone may need revision.
How secure are Australian emergency departments?
Authors: Merfield Eve
Published: November 2003

Objective: To ascertain the level of security arrangements in Australian EDs.

Methods: A postal survey to the directors of all Australasian College for Emergency Medicine (ACEM) accredited EDs in Australia. A score based on the ACEM design guidelines for EDs was given. A maximum score of 12 correlated with compliance with the 12 specific ACEM recommendations. Questions regarding satisfaction with security and occurrence of a major incident within the prior five years were included.

Results: There was an 88% response rate to the questionnaire. Of the respondents, 63% were unhappy with the level of security in their ED and 81% reported the occurrence of a major incident within five years. The average score out of 12 was 8.39 (SD 2.4). A total of 7% scored 12/12 and 21% scored 6 or less. There was no difference in scores when the ED were grouped by age, locality or annual attendances.

Conclusions: The vast majority of Australian EDs do not comply with all ACEM recommendations on security and most department directors are unhappy with the existing level of security in their departments. This is of concern in light of the negative impact of violence on the well‐being of ED staff.
AUTOLOGOUS FAT INJECTION LARYNGOPLASTY FOR UNILATERAL VOCAL FOLD PARALYSIS
Authors: Havas Thomas E. , Priestley K. Jocelyn
Published: November 2003

Background:  Fat injection laryngoplasty has been used at the Sydney Voice Clinic for selected cases of unilateral vocal fold paralysis since 1989.

Methods:  Forty‐five consecutive cases deemed suitable for treatment by this technique are presented in this paper.

Results:  Mean follow up for this group of patients was 33 months. Over the period of follow up, 39 of the 45 patients achieved normal or near normal voice, with four patients requiring additional surgical intervention.

Conclusion:  Fat injection laryngoplasty is a quick, simple, inexpensive and reliable procedure, with few complications and good long‐term results in suitable selected cases of unilateral vocal fold paralysis.