Plasmodium knowlesi is an intracellular malaria parasite whose natural vertebrate host is Macaca fascicularis (the ‘kra’ monkey); however, it is now increasingly recognized as a significant cause of human malaria, particularly in southeast Asia1,2. Plasmodium knowlesi was the first malaria parasite species in which antigenic variation was demonstrated3, and it has a close phylogenetic relationship to Plasmodium vivax4, the second most important species of human malaria parasite (reviewed in ref. 4). Despite their relatedness, there are important phenotypic differences between them, such as host blood cell preference, absence of a dormant liver stage or ‘hypnozoite’ in P. knowlesi, and length of the asexual cycle (reviewed in ref. 4). Here we present an analysis of the P. knowlesi (H strain, Pk1(A+) clone5) nuclear genome sequence. This is the first monkey malaria parasite genome to be described, and it provides an opportunity for comparison with the recently completed P. vivax genome4 and other sequenced Plasmodium genomes6-8. In contrast to other Plasmodium genomes, putative variant antigen families are dispersed throughout the genome and are associated with intrachromosomal telomere repeats. One of these families, the KIRs9, contains sequences that collectively match over one-half of the host CD99 extracellular domain, which may represent an unusual form of molecular mimicry.
The dissociation constants for the binding of Rhodobacter capsulatus cytochrome c2 and its K93P mutant to the cytochrome bc1 complex embedded in a phospholipid bilayer were measured by plasmon waveguide resonance spectroscopy in the presence and absence of the inhibitor stigmatellin. The reduced form of cytochrome c2 strongly binds to reduced cytochrome bc1 (Kd = 0.02 μM), but binds much more weakly to the oxidized form (Kd = 3.1 μM). In contrast, oxidized cytochrome c2 binds to oxidized cytochrome bc1 in a biphasic fashion with Kd values of 0.11 μM and 0.58 μM. Such a biphasic interaction is consistent with binding to two separate sites or conformations of oxidized cytochrome c2 and/or cytochrome bc1. However, in the presence of stigmatellin, we find that oxidized cytochrome c2 binds to oxidized cytochrome bc1 in a monophasic fashion with high affinity (Kd = 0.06 μM) and reduced cytochrome c2 binds less strongly (Kd = 0.11 μM) but ∼30 fold more tightly than in the absence of stigmatellin. Structural studies with cytochrome bc1, with and without the inhibitor stigmatellin, have lead to the proposal that the Rieske protein is mobile, moving between the cytochrome b and cytochrome c1 components during turnover. In one conformation, the Rieske protein binds near the heme of cytochrome c1, while the cytochrome c2 binding site is also near the cytochrome c1 heme but on the opposite side from the Rieske site, where cytochrome c2 cannot directly interact with Rieske. However, the inhibitor, stigmatellin, freezes the Rieske protein iron-sulfur cluster in a conformation proximal to cytochrome b and distal to cytochrome c1. We conclude from this that the dual conformation of the Rieske protein is primarily responsible for biphasic binding of oxidized cytochrome c2 to cytochrome c1. This optimizes turnover by maximizing binding of the substrate, oxidized cytochrome c2, when the iron-sulfur cluster is proximal to cytochrome b and minimizing binding of the product, reduced cytochrome c2, when it is proximal to cytochrome c1.
Stigmatellin; protein-protein interaction; binding constants; structural changes
Ruminant animals are carriers of Escherichia coli O157:H7, and the transmission of E. coli O157:H7 from cattle to the environment and to humans is a concern. It is unclear if diet can influence the survivability of E. coli O157:H7 in the gastrointestinal system or in feces in the environment. Feces from cattle fed bromegrass hay or corn silage diets were inoculated with E. coli O157:H7, and the survival of this pathogen was analyzed. When animals consumed bromegrass hay for <1 month, viable E. coli O157:H7 was not recovered after 28 days postinoculation, but when animals consumed the diet for >1 month, E. coli O157:H7 cells were recovered for >120 days. Viable E. coli O157:H7 cells in feces from animals fed corn silage were detected until day 45 and differed little with the time on the diet. To determine if forage phenolic acids affected the viability of E. coli O157:H7, feces from animals fed corn silage or cracked corn were amended with common forage phenolic acids. When 0.5% trans-cinnamic acid or 0.5% para-coumaric acid was added to feces from silage-fed animals, the E. coli O157:H7 death rate was increased significantly (17-fold and 23-fold, respectively) compared to that with no addition. In feces from animals fed cracked corn, E. coli O157:H7 death rates were increased significantly with the addition of 0.1% and 0.5% trans-cinnamic acid (7- and 13-fold), 0.1% and 0.5% p-coumaric acid (3- and 8-fold), and 0.5% ferulic acid (3-fold). These data suggest that phenolic acids common to forage plants can decrease viable counts of E. coli O157:H7 shed in feces.
BACKGROUND—Endoscopic ultrasound (EUS) may be used for preoperative staging of gastro-oesophageal carcinoma but performance values given in the literature differ.
AIMS—To identify and synthesise findings from all articles on the performance of EUS in tumour, node, metastasis (TNM) staging of gastro-oesophageal carcinoma.
SOURCE—Published and unpublished English language literature, 1981-1996.
METHODS—Data on the staging performance of EUS were retrieved and evaluated. Summary receiver operator characteristic methodology was used for synthesis, and a summary estimate of performance, Q*, obtained. Multiple regression analysis was used to assess study validity and investigate reasons for differences in performance.
RESULTS—Twenty seven primary articles were assessed in detail. Thirteen supplied results for staging oesophageal cancer, 13 for gastric cancer, and four for cancers at the gastro-oesophageal junction. For gastric T staging, Q*=0.93 (95% confidence interval (CI) 0.91-0.95) and for oesophageal T staging, Q*=0.89 (95% CI 0.88-0.92). For gastro-oesophageal T staging, including cancers at the gastro-oesophageal junction, Q*=0.91 (95% CI 0.89-0.93). Inclusion of cases with non-traversable stenosis was found to slightly reduce staging performance. For N staging, Q*=0.79 (95% CI 0.75-0.83). In articles that compared EUS directly with incremental computed tomography, EUS performed better. None of the variables assessed in the regression analysis was significant using a Bonferroni correction. Three variables (anatomical location, traversability, and blinding) showed strong relationships for future research and validation.
CONCLUSIONS—EUS is highly effective for discrimination of stages T1 and T2 from stages T3 and T4 for primary gastro-oesophageal carcinomas. The failure rate of EUS from non-traversability of a stenotic cancer may be a limitation in some patient groups.
Keywords: endoscopic ultrasound; gastro-oesophageal cancer; TNM staging; systematic literature review; meta-analysis
Laser safety analysis; medical imaging; non-ionizing radiation; perception of risk; terahertz pulsed imaging; thermal effects
Modelling the interaction of terahertz(THz) radiation with biological tissueposes many interesting problems. THzradiation is neither obviously described byan electric field distribution or anensemble of photons and biological tissueis an inhomogeneous medium with anelectronic permittivity that is bothspatially and frequency dependent making ita complex system to model.A three-layer system of parallel-sidedslabs has been used as the system throughwhich the passage of THz radiation has beensimulated. Two modelling approaches havebeen developed a thin film matrix model anda Monte Carlo model. The source data foreach of these methods, taken at the sametime as the data recorded to experimentallyverify them, was a THz spectrum that hadpassed though air only.Experimental verification of these twomodels was carried out using athree-layered in vitro phantom. Simulatedtransmission spectrum data was compared toexperimental transmission spectrum datafirst to determine and then to compare theaccuracy of the two methods. Goodagreement was found, with typical resultshaving a correlation coefficient of 0.90for the thin film matrix model and 0.78 forthe Monte Carlo model over the full THzspectrum. Further work is underway toimprove the models above 1 THz.
Biological tissue; modelling; Monte Carlo; simulation; skin; terahertz
Recently published studies suggest thatterahertz pulsed imaging will have applications inmedicine and biology, but there iscurrently very little information about the opticalproperties of human tissue at terahertzfrequencies. Such information would be useful forpredicting the feasibility of proposedapplications, optimising acquisition protocols,providing information about variability ofhealthy tissue and supplying data for studies of theinteraction mechanisms. Research ethicscommittee approval was obtained, andmeasurements made from samples of freshlyexcised human tissue, using a broadbandterahertz pulsed imaging system comprisingfrequencies approximately 0.5 to 2.5 THz.Refractive index and linear absorptioncoefficient were found. Reproducibility wasdetermined using blood from one volunteer,which was drawn and measured on consecutivedays. Skin, adipose tissue, striatedmuscle, vein and nerve were measured (to date, from oneindividual). Water had a higher refractiveindex (2.04 ± 0.07) than any tissue.The linear absorption coefficient was higher formuscle than adipose tissue, as expectedfrom the higher hydration of muscle. As these samples camefrom a single subject, there is currentlyinsufficient statistical power to draw firmconclusions, but results suggest that in vivo clinical imaging will be feasible in certainapplications.
absorption; human tissue; medical imaging; refractive index; terahertz pulsed imaging
OBJECTIVES—CD4+ T cells sustain the chronic synovial inflammatory response in rheumatoid arthritis (RA). SB-210396/CE 9.1 is an anti-CD4 monoclonal antibody that has documented efficacy in RA when given intravenously. This study aimed to establish the safety and efficacy of the intra-articular administration of SB-210396/CE 9.1 compared with placebo, examining its mode of action using a combined imaging approach of arthroscopy, magnetic resonance imaging (MRI), and histology.
METHODS—Thirteen RA patients with active, resistant knee synovitis, were randomised to intra-articular injection of placebo (n=3), 0.4 mg (n=3) or 40 mg (n=7) of anti-CD4 after sequential dynamic gadolinium enhanced MRI, followed by same day arthroscopy and synovial membrane biopsy. Imaging and arthroscopic synovial membrane sampling were repeated at six weeks. This study used a unique region of interest (ROI) analysis mapping the MRI area analysed to the specific biopsy site identified arthroscopically, thus providing data for all three modalities at the same synovial membrane site.
RESULTS—12 patients completed the study (one placebo treated patient refused further MRI). Arthroscopic improvement was observed in 0 of 2 placebo patients but in 10 of 10 patients receiving active drug (>20% in 6 of 10). Improvement in MRI was consistently observed in all patients of the 40 mg group but not in the other two groups. A reduction in SM CD4+ score was noted in the 40 mg group and in the 0.4 mg group. Strong correlations both before and after treatment, were identified between the three imaging modalities. Intra-articular delivery of SB-210396/CE 9.1 was well tolerated.
CONCLUSIONS—SB-210396/CE 9.1 is safe when administered by intra-articular injection. A trend toward efficacy was found by coordinated MRI, arthroscopic, and histological imaging, not seen in the placebo group. The value of ROI analysis was demonstrated.
Adherence to hydroxyapatite (HA) was examined as a method to concentrate bacteria from foods. Using HA at a level of 10% and suspensions of an Escherichia coli strain containing 10(9), 10(6), and 10(3) cells per ml, kinetic studies revealed that maximum adherence was attained within 5 min for all cell concentrations and that comparable log reductions (1.0 to 1.5) of cells in suspension were seen regardless of initial cell concentration. Eleven species of spoilage and pathogenic bacteria were found to adhere to HA, with seven species adhering at proportions of greater than 95%. Fluorescent viability staining revealed that cells bound to HA remained viable. There was greater than 92% adherence of indigenous bacteria to HA from three of five 1:10 dilutions of ground beef, indicating promise for the use of HA for concentrating bacteria from meat and other food samples.
We have developed a model system to assess the influence of earthworm activity on the transfer of plasmid pJP4 from an inoculated donor bacterium, Pseudomonas fluorescens C5t (pJP4), to indigenous soil microorganisms. Three different earthworm species (Lumbricus terrestris, Lumbricus rubellus, and Aporrectodea trapezoides), each with unique burrowing, casting, and feeding behaviors, were evaluated. Soil columns were inoculated on the surface with 10(8) cells per g of soil of the donor bacterium, and after a 2-week incubation period, donor, transconjugant, and total bacteria were enumerated at 5-cm-depth intervals. Transconjugants were confirmed by use of colony hybridization with a mer gene probe. In situ gene transfer of plasmid pJP4 from P. fluorescens C5t to indigenous soil bacteria was detected in all inoculated microcosms. In the absence of earthworms, the depth of recovery was limited to the top 5 cm of the column, with approximately 10(3) transconjugants per g of soil. However, the total number of transconjugants recovered from soil was significantly greater in microcosms containing either L. rubellus or A. trapezoides, with levels reaching about 10(5) CFU/g of soil. In addition, earthworms distributed donor and transconjugant bacteria throughout the microcosm columns, with the depth of recovery dependent on the burrowing behavior of each earthworm species. Donor and transconjugant bacteria were also recovered from earthworm casts and inside developing cocoons. Transconjugant bacteria from the indigenous soil microflora were classified as belonging to Acidovorax spp., Acinetobacter spp., Agrobacterium spp., Pasteurella spp., Pseudomonas spp., and Xanthomonas spp.
Previous studies indicated that dispersal of S. carpocapsae may be enhanced in soil with earthworms. The objective of this research was to determine and compare the effects of earthworms on dispersal of other Steinernema spp. Vertical dispersal of Steinernema carpocapsae, S. feltiae, and S. glaseri was tested in soil columns in the presence and absence of earthworms (Lumbricus terrestris). Dispersal was evaluated by a bioassay and by direct extraction of nematodes from soil. Upward dispersal of S. carpocapsae and S. feltiae increased in the presence of earthworms, whereas upward dispersal of S. glaseri was not affected by earthworms. No significant differences were detected in downward dispersal of S. carpocapsae and S. feltiae in soil with earthworms compared to soil without earthworms. Downward dispersal of S. glaseri, however, was greater in soil without earthworms relative to soil with earthworms. In soil void of earthworms, dispersal of S. glaseri was greatest followed by dispersal of S. carpocapsae. The presence of earthworm burrows in soil did not influence nematode dispersal. Nematodes were recovered from the surface, interior, and casts of earthworms. Therefore, nematodes may have a phoretic association with earthworms.
earthworm; entomophyllic nematode; nematode; nematode dispersal; Steinernema spp.
The relationship between tumour growth, insulin status, blood lipids and adipose linoleic acid (LA, reflecting long-term LA intake) was studied in 19 Jewish women suffering from early and advanced stages (ES and AS) of ovarian and endometrial tumours. Blood insulin in patients with ES tumours was four times higher than the control value in cancer-free subjects, but fell to normal levels at AS and after ES surgery (PES). Tumours and abdominal adipose tissue (AAT) had 4-6 and 1.4-1.7 times as much insulin as non-cancerous control organs. Serum total cholesterol (CHOL) and LDL-cholesterol were high at ES, dropped below normal at AS, but normalised at PES, while HDL-cholesterol increased after ES surgery. Linoleic acid in subcutaneous adipose tissue (SAT) was high in controls (26.4 + 1.5% of total fatty acids), but lower in cancer patients (20.5 + 3.7%, P < 0.05), while palmitic acid showed the opposite change. The results suggest mobilisation of glucose, cholesterol and linoleic acid for the supply of energy and structural lipids to rapidly multiplying tumour cells and possibly for prostaglandin synthesis. They also raise the question of whether the high linoleic acid intake by the Jewish population in Israel predisposes individuals to tumour development.
Dispersal of the nematode Steinernema carpocapsae (All strain), applied on the top or the bottom of soil columns, was tested in the presence or absence of two earthworm species, Lumbricus terrestris or Aporrectodea trapezoides. Nematode dispersal was estimated after a 2-week period with a bioassay against the greater wax moth, Galleria mellonella. Vertical dispersal of nematodes was increased in the presence of earthworms. When nematodes were placed on the surface of soil columns, significantly more nematodes dispersed to the lower half of the columns when either earthworm species was present than when earthworms were not present. When nematodes were placed on the bottom of soil columns, significantly more nematodes dispersed to the upper half of the columns when L. terrestris was present than when A. trapezoides was present or in the absence of earthworms. Because nematodes were found on the exterior and in the interior of earthworms, nematode dispersal may be enhanced by direct contact with the earthworms.
dispersal; earthworm; nematode; nematode dispersal; Steinernema carpocapsae; Steinernematidae
A standard lyophilised triple cryoprecipitate preparation, stabilised by the addition of Synthamin 17, was heat treated at 60 degrees C for 48 hours. The total protein content, factor VIII concentration, and factor VIII recovery were not affected by the heat treatment procedure. Heat treatment did not influence the reconstitution characteristics of the freeze dried preparation and there were no side effects during or after administration. The mean in vivo rise of factor VIII from infused heat treated triple cryoprecipitate was 2.5 (SD 0.9)%/unit/kg with a half life of 13.1 (3.1) hours. These results compare favourably with those obtained using non-heated triple cryoprecipitate. Cryoprecipitate can be heat treated without adversely influencing factor VIII recovery, and the ability to prepare a heat treated cryoprecipitate means that a small pool high yield factor VIII preparation can again be used in routine clinical practice.
Interferon production was demonstrated by the goldfish-derived CAR cell line in response to infection by goldfish virus-2. Supernatants of infected cultures provided antiviral protection to CAR cells and another cell line derived from goldfish, ABIII. The protective factor retained activity after ultracentrifugation, dialysis, freezing and thawing, acid treatment (pH 2), or heating to 56 degrees C but was sensitive to trypsin. Supernatants of infected cultures did not affect adsorption of virus. Previous studies have shown that replication of goldfish virus type 2 is enhanced by pretreatment of cultures with subcytotoxic concentrations of carbaryl. In the present study, pesticide-treated cultures were found to synthesize reduced levels of interferon.
1 The effect of the surfactant, Cetomacrogol 1000, on the absorption of insulin across the rectal mucosa has been studied. 2 Rectal administration of microenemata containing Cetomacrogal 1000 and insulin causes a rise in the plasma concentration of insulin and a consequent fall in the blood glucose concentration in diabetic and non-diabetic rats. 3 The hypoglycaemic response is dependent on both the concentration of surfactant and the dose of insulin administered. 4 The results suggest that the transport of insulin across the rectal mucosa is facilitated by Cetomacrogal 1000.
A clinical report of a 30-year-old woman who developed acute hepatic failure in the fifth month of pregnancy is presented. L-dopa administration resulted in a marked improvement in both level of consciousness and electroencephalogram. The literature dealing with L-dopa therapy in hepatic encephalopathy is reviewed.
Three patients with fever and malaise, one of whom also had joint pains, were extensively investigated before their condition was attributed to dental sepsis. Each patient recovered fully after appropriate dental treatment. Dental sepsis should be added to the list of possible causes of pyrexia of undetermined origin, and a routine dental examination should be carried out in each case.
Waterworth, Pamela M., Lockey, Eunice, Berry, E. M., and Pearce, Helen M. (1974).Thorax, 29, 432-436. A critical investigation into the antibiotic sterilization of heart valve homografts. Sterility tests of homograft heart valves sterilized by antibiotics were found to be invalidated by carry-over of the antibiotics. A method was devised to test the efficacy of different antibiotic mixtures using membrane filters. Fifty-five organisms, including 13 species, were tested, and sterility was achieved with only 35.
The relationship of plasma calcium levels to changes in plasma specific gravity, total protein, and albumin induced by venous stasis was investigated. Factors were derived for adjusting calcium results to offset the effects of variation in protein concentration and thus to make them of increased discriminatory value to the clinician. The validity of an existing specific gravity correction has been substantiated, but a more exact adjustment of 0·23 mg/100 ml of calcium for every 0·001 change in specific gravity is proposed. We recommend for automated laboratories that the factor based on albumin be used: 0·09 mg/100 ml of calcium should be subtracted from the total calcium value for every increase of 0·1 g in albumin above 4·6 g/100 ml, and a corresponding addition should be made for values of albumin below 4·6 g/100 ml.
Using a calcium specific electrode, it has been shown that the ionized calcium concentration does not alter with prolonged venous stasis.
Dietary triacylglycerols containing palmitic acid in the sn-2 position might impair insulin release and increase plasma glucose.
We used a cross-over designed feeding trial in 53 healthy Asian men and women (20–50 years) to test this hypothesis by exchanging 20% energy of palm olein (PO; control) with randomly interesterified PO (IPO) or high oleic acid sunflower oil (HOS). After a 2-week run-in period on PO, participants were fed PO, IPO and HOS for 6 week consecutively in randomly allocated sequences. Fasting (midpoint and endpoint) and postprandial blood at the endpoint following a test meal (3.54 MJ, 14 g protein, 85 g carbohydrate and 50 g fat as PO) were collected for the measurement of C-peptide, insulin, glucose, plasma glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1, lipids and apolipoproteins; pre-specified primary and secondary outcomes were postprandial changes in C-peptide and plasma glucose.
Low density lipoprotein cholesterol was 0.3 mmol/l (95% confidence interval (95% CI)) 0.1, 0.5; P<0.001) lower on HOS than on PO or IPO as predicted, indicating good compliance to the dietary intervention. There were no significant differences (P=0.58) between diets among the 10 male and 31 female completers in the incremental area under the curve (0–2 h) for C-peptide in nmol.120 min/l: GM (95% CI) were PO 220 (196, 245), IPO 212 (190, 235) and HOS 224 (204, 244). Plasma glucose was 8% lower at 2 h on IPO vs PO and HOS (both P<0.05).
Palmitic acid in the sn-2 position does not adversely impair insulin secretion and glucose homeostasis.
Within the limits of this study, it was found that 5 ppm of cobalt was adequate to give good levels of vitamin B12. The vitamin B12 precursor 5,6-dimethylbenzimidazole was found to be adequate at 10 ppm in the absence of aeration. In the presence of aeration, a zero level of precursor was found to be most desirable. The analysis of variance showed aeration to be highly significant, and the aeration and precursor interaction to be significant. No other significant effects were observed.
The patterns of growth and vitamin formation by Propionibacterium shermanii in whey were similar to the patterns established in other substrates. The vitamin formation was observed during the latter part of the fermentation after the organism approached maximal growth. Lactose utilization by the organism corresponded to the logarithmic-growth phase of the organism. Analyses of the dried culture showed a large increase of vitamin B12 in the fermentation solids compared with unfermented dried whey. A feed analysis showed a notable increase of protein and a large decrease in nitrogen-free extract of the dried fermentation solid compared with dried whey.