Cost and effectiveness comparison of two methods for screening potential blood donors for anaemia in Vietnam.

AIM/OBJECTIVE: To compare the cost and effectiveness of Copper Sulphate (CS) and HemoCue (HC) methods for screening blood donors for anaemia. BACKGROUND: Robust information from developing countries about cost and effectiveness of anaemia screening methods for blood donors is scarce. In such countries there are widespread shortages of blood, so the most cost-effective method should maximise blood supply without compromising donor safety. METHODS: Economic data (e.g. staff time, equipment and buildings) were collected from direct observation of procedures and purchase data from Hanoi's Central Blood Bank administrative department. A framework for comparing the cost and effectiveness of anaemia screening methods was developed and a cost per effective (i.e. usable and accurate) test was generated for each method. RESULTS: Samples from 100 potential donors from the Hanoi Central Blood Bank (static) and 198 from two mobile units were tested. The mean probability of an ineffective anaemia test was 0·1 (0·05-0·2). The average cost of an HC test was $0·75 (static $0·61 and mobile $0·89) and a CS test was $0·31 (static $0·17 and mobile $0·45). The difference between static and mobile units was predominantly due to transport costs; the difference between the two methods was predominantly due to the HC microcuvettes. CONCLUSION: In this setting the CS yields greater value for money than the HC method for screening blood donors. The relative cost and effectiveness of CS and HC may be different in places with higher staff turnover, lower test accuracy, higher anaemia prevalence or lower workload than in Vietnam.

An experimental study of the putative mechanism of a synthetic autonomous rotary DNA nanomotor.

DNA has been used to construct a wide variety of nanoscale molecular devices. Inspiration for such synthetic molecular machines is frequently drawn from protein motors, which are naturally occurring and ubiquitous. However, despite the fact that rotary motors such as ATP synthase and the bacterial flagellar motor play extremely important roles in nature, very few rotary devices have been constructed using DNA. This paper describes an experimental study of the putative mechanism of a rotary DNA nanomotor, which is based on strand displacement, the phenomenon that powers many synthetic linear DNA motors. Unlike other examples of rotary DNA machines, the device described here is designed to be capable of autonomous operation after it is triggered. The experimental results are consistent with operation of the motor as expected, and future work on an enhanced motor design may allow rotation to be observed at the single-molecule level. The rotary motor concept presented here has potential applications in molecular processing, DNA computing, biosensing and photonics.

Isolation of a human tissue-type plasminogen-activator genomic DNA clone and its expression in mouse L cells.

We have isolated a cDNA clone corresponding to a substantial portion of the human tissue-type plasminogen activator (t-PA) protein. It encodes almost all of the protein B chain and part of the 3' untranslated region. We have used this clone to screen bacteriophage lambda and cosmid libraries of human genomic DNA. Several related genomic clones were isolated. One of these, a cosmid clone, carried approx. 40 kb of human DNA. Mapping experiments indicate that the region containing the protein-coding exons is approx. 20 kb in length. The cosmid, containing the t-PA gene and the aminoglycosyl-3'-phosphotransferase dominant-selection marker, was introduced into mouse L cells. Approximately half of the transformants were shown to produce human t-PA. We demonstrated that the fibrinolytic t-PA activity could be specifically quenched by anti-t-PA antibody and that the recombinant t-PA was of similar size (by SDS-polyacrylamide gel electrophoresis) to the t-PA produced by the human Bowes melanoma cell line. Our results suggest that the cosmid clone carries the whole t-PA coding region together with the regulatory elements necessary for its expression.

Inhibitors of blood platelet aggregation. Effects of some 1,2-benzisothiazol-3-ones on platelet responsiveness to adenosine diphosphate and collagen.

A series of substituted 1,2-benzisothiazol-3-ones was synthesized, and the compounds were tested for ability to inhibit platelet aggregation induced by adenosine diphosphate and collagen in rats and guinea pigs ex vivo. Alkyl substituents at the 2-position bearing a basic group were necessary for ex vivo activity. Several of the compounds were potent inhibitors of adenosine diphosphate induced first-phase aggregation, but adverse toxicological findings terminated their further development. Preliminary studies suggested that inhibition of aggregation was not attributable to inhibition of prostanoid synthesis or to raised levels of cyclic 3',5'-adenosine monophosphate.

Effects of novel bile salts on cholesterol metabolism in rats and guinea-pigs.

Novel bile salts (quaternary ammonium conjugates) inhibited cholic acid binding and transport in everted ileal sacs in vitro. The cationic piperazine conjugate of lithocholic acid (di-iodide salt, compound 8, BRL 39924A) appeared most active, inhibiting binding by 29% and transport by 59% in guinea-pig ileum (200 microM). BRL 39924A also inhibited taurocholate uptake into guinea-pig ileal sacs and cholate uptake into rat ileal sacs and was selected for further study in vivo. In hyperlipidaemic rats, BRL 39924A significantly raised cholesterol 7 alpha-hydroxylase activity and decreased hepatic accumulation of exogenous cholic acid. HDL cholesterol concentration in the serum increased and the level of VLDL plus LDL cholesterol decreased. In hyperlipidaemic guinea-pigs. BRL 39924A lowered serum total cholesterol and triglyceride levels. Although metabolic changes were less than those achieved with the bile acid sequestrant, cholestyramine, the doses of BRL 39924A used were much lower (100-500 mg/kg body wt). Selective inhibition of receptor mediated bile acid uptake may be associated with local side-effects but these novel bile salts are useful pharmacological tools to examine the effects of receptor blockade on lipoprotein metabolism.

Circadian variation in human stature.

The aim of this study was to monitor the creep in stature due to compression and its recovery over 24 hr in eight adult males. Measurements of stature were made at nine times during the 24-hr cycle using a purpose built metal frame tilted 5 degrees to the vertical. Accessories for standardization of posture and prevention of unwanted muscular tension included a series of microswitches on the frame, cross-beams for controlling spinal curvatures, slit spectacles used in conjunction with a mirror for proper head alignment. A dead load BAYE micrometer recorded stature to 0.01 mm. A significant circadian rhythm was established, the trough to peak variation being 19.3 mm or 1.1% of overall stature. Peak stature was measured at 0730 on awakening and the trough occurred at midnight before assuming a recumbent posture for sleep. Altogether 71% of the height gained during the night was achieved in the first half of the night's sleep. Over 50% of the height loss in a day was lost within the first hour of rising, 80% being lost within 3 hr of arising: the rate of creep decelerated throughout the remainder of the waking day. It is concluded that the rate of change in creep throughout the day varies, being greatest in the morning whilst distension is most pronounced in the early hours of sleep.

Circadian variation in stature and the effects of spinal loading.

Using a method comparable with that of Eklund and Corlett (1984) stature was measured with an accuracy of I mm in eight young adults. The mean circadian variation was 19.3 mm (1.1% of stature). Fifty-four percent of the diurnal loss in stature occurred in the first hour after rising. Approximately 70% was regained during the first half of the night. With static shoulder loads (2.5-40 kg), increases in the rate of shrinkage with increasing weight were nonlinear. Repetitive lifting led to greater shrinkage than with equivalent static loading. Rest in Fowler's position gave more rapid regains in stature than post-exercise recovery in standing positions. The technique is therefore suitable for assessment of the effects of manual work with both occupational and therapeutic applications.

The current status of nutrition labelling in the United Kingdom.

During the past 10 years there has been considerable discussion about nutrition labelling of foodstuffs. In the United Kingdom, the Ministry of Agriculture, Fisheries and Food has published several proposals for guidelines and regulations. The European Community has more recently considered this subject and has issued their proposals for draft directives. There is now considerable agreement between the UK guidelines on nutrition labelling published in 1987 and EC proposals which are under discussion. Compulsory nutrition labelling is unlikely in the immediate future, except when a nutrition claim is made, when labelling is to be compulsory and must conform to one of several specified formats. In contrast, many consumer groups believe that nutrition labelling should be compulsory for all packaged foods. The Ministry of Agriculture, Fisheries and Food intends to control nutrition claims, such as claims of 'high' or 'low' levels of various nutrients, and definitions of these nutrient levels have been proposed for discussion. Whether to label foods according to 'total' or 'added' sugars content, or both, is also under discussion.

A survey of nutrition labelling of sugar-containing foods in the north of England in 1989.

A survey of the nutrition labelling of 880 varieties of foods on sale in three stores in Newcastle upon Tyne was undertaken in May-July 1989. The foods were chosen for investigation because they contained sugars, and they were categorised into 12 types of food. Some nutritional information was given for most foods but it seldom conformed to the format suggested by the Ministry of Agriculture, Fisheries and Food. The sugars content of the foods was seldom given. Even when nutritional claims were made (e.g. 'low sugar', 'high fibre'), nutritional information was often incomplete. The three stores differed in the extent to which their own-brand products were labelled for nutrient content. Tesco products were comprehensibly labelled, while own-brand products for sale in the other two stores were not, although both stores stated their intention to introduce comprehensive nutrition labelling. Two of the three stores had, or intended to have, obligatory sugar labelling. Despite this, there were many examples of nutrition labelling which was misleading. It is concluded that nutrition labelling should be compulsory and should conform to a format which specifies sugars content.

Forensic three-dimensional facial reconstruction: historical review and contemporary developments.

Despite varied attempts to achieve standardization in traditional techniques and the promotion of some newly developed ones, facial reconstruction remains on the threshold between art and science. It is the point at which science ends and the medical illustrator takes over that has led to most reservations over this branch of forensic anthropology. The purpose of this paper is to demonstrate that many techniques of facial reconstruction are prima facie questionable and to illustrate some possible solutions to the problems which are currently being explored by the Facial Reconstruction Project at the University of Sheffield (UK). The review includes 15 responses to a questionnaire which was offered to facial reconstruction experts and related specialists. The use of 3D color laser scanning equipment, collection of tissue depth measurements from CT scans and the development of a computer system for 3D forensic facial reconstruction, are described.

Abdominal aortic aneurysm: diagnosis, treatment, and implications for advanced practice nursing.

The nurse practitioner needs to acknowledge that not all patients are insightful. The patient who is naive, stoical, or in denial may not return to his previous level of health. Behavioral or environmental changes may be a necessary part of recovery. The advanced practice nurse, through her research and subsequent knowledge, can identify and implement holistic changes necessary for the maintenance of health and the development of appropriate health-seeking behaviors that lower the morbidity and mortality for such conditions as abdominal aortic aneurysms. Nurse practitioners play a vital role in research, prevention and early detection of major threats to wellness (Lawler and Schmidt, 1992). Gender-sensitive research regarding factors affecting recovery are also necessary as females respond differently to such conditions as renal failure (Carlson and Eisenstat, 1995).

On the differences between conventional and auditory spectrograms of English consonants.

A new tool for speech analysis is presented, operating in real-time and incorporating the analysing power of a contemporary auditory model to produce the familiar display of the speech spectrograph. This "auditory spectrograph" is used to analyse English consonant sounds and the results are compared with conventional wide and narrow band spectrograms. The auditory analyses are found to attach more visual weight to the acoustic cues associated with speech production and perception, and features that are either difficult or impossible to distinguish on conventional spectrograms are clarified.

SABRE: a bio-inspired fault-tolerant electronic architecture.

As electronic devices become increasingly complex, ensuring their reliable, fault-free operation is becoming correspondingly more challenging. It can be observed that, in spite of their complexity, biological systems are highly reliable and fault tolerant. Hence, we are motivated to take inspiration for biological systems in the design of electronic ones. In SABRE (self-healing cellular architectures for biologically inspired highly reliable electronic systems), we have designed a bio-inspired fault-tolerant hierarchical architecture for this purpose. As in biology, the foundation for the whole system is cellular in nature, with each cell able to detect faults in its operation and trigger intra-cellular or extra-cellular repair as required. At the next level in the hierarchy, arrays of cells are configured and controlled as function units in a transport triggered architecture (TTA), which is able to perform partial-dynamic reconfiguration to rectify problems that cannot be solved at the cellular level. Each TTA is, in turn, part of a larger multi-processor system which employs coarser grain reconfiguration to tolerate faults that cause a processor to fail. In this paper, we describe the details of operation of each layer of the SABRE hierarchy, and how these layers interact to provide a high systemic level of fault tolerance.

Transcriptional profiling of the bladder in urogenital schistosomiasis reveals pathways of inflammatory fibrosis and urothelial compromise.

Urogenital schistosomiasis, chronic infection by Schistosoma haematobium, affects 112 million people worldwide. S. haematobium worm oviposition in the bladder wall leads to granulomatous inflammation, fibrosis, and egg expulsion into the urine. Despite the global impact of urogenital schistosomiasis, basic understanding of the associated pathologic mechanisms has been incomplete due to the lack of suitable animal models. We leveraged our recently developed mouse model of urogenital schistosomiasis to perform the first-ever profiling of the early molecular events that occur in the bladder in response to the introduction of S. haematobium eggs. Microarray analysis of bladders revealed rapid, differential transcription of large numbers of genes, peaking three weeks post-egg administration. Many differentially transcribed genes were related to the canonical Type 2 anti-schistosomal immune response, as reflected by the development of egg-based bladder granulomata. Numerous collagen and metalloproteinase genes were differentially transcribed over time, revealing complex remodeling and fibrosis of the bladder that was confirmed by Masson's Trichrome staining. Multiple genes implicated in carcinogenesis pathways, including vascular endothelial growth factor-, oncogene-, and mammary tumor-related genes, were differentially transcribed in egg-injected bladders. Surprisingly, junctional adhesion molecule, claudin and uroplakin genes, key components for maintaining the urothelial barrier, were globally suppressed after bladder exposure to eggs. This occurred in the setting of urothelial hyperplasia and egg shedding in urine. Thus, S. haematobium egg expulsion is associated with intricate modulation of the urothelial barrier on the cellular and molecular level. Taken together, our findings have important implications for understanding host-parasite interactions and carcinogenesis in urogenital schistosomiasis, and may provide clues for novel therapeutic strategies.

Stable isotope probing with 15N achieved by disentangling the effects of genome G+C content and isotope enrichment on DNA density.

Stable isotope probing (SIP) of nucleic acids is a powerful tool that can identify the functional capabilities of noncultivated microorganisms as they occur in microbial communities. While it has been suggested previously that nucleic acid SIP can be performed with 15N, nearly all applications of this technique to date have used 13C. Successful application of SIP using 15N-DNA (15N-DNA-SIP) has been limited, because the maximum shift in buoyant density that can be achieved in CsCl gradients is approximately 0.016 g ml-1 for 15N-labeled DNA, relative to 0.036 g ml-1 for 13C-labeled DNA. In contrast, variation in genome G+C content between microorganisms can result in DNA samples that vary in buoyant density by as much as 0.05 g ml-1. Thus, natural variation in genome G+C content in complex communities prevents the effective separation of 15N-labeled DNA from unlabeled DNA. We describe a method which disentangles the effects of isotope incorporation and genome G+C content on DNA buoyant density and makes it possible to isolate 15N-labeled DNA from heterogeneous mixtures of DNA. This method relies on recovery of "heavy" DNA from primary CsCl density gradients followed by purification of 15N-labeled DNA from unlabeled high-G+C-content DNA in secondary CsCl density gradients containing bis-benzimide. This technique, by providing a means to enhance separation of isotopically labeled DNA from unlabeled DNA, makes it possible to use 15N-labeled compounds effectively in DNA-SIP experiments and also will be effective for removing unlabeled DNA from isotopically labeled DNA in 13C-DNA-SIP applications.

Stable isotope probing with 15N2 reveals novel noncultivated diazotrophs in soil.

Biological nitrogen fixation is a fundamental component of the nitrogen cycle and is the dominant natural process through which fixed nitrogen is made available to the biosphere. While the process of nitrogen fixation has been studied extensively with a limited set of cultivated isolates, examinations of nifH gene diversity in natural systems reveal the existence of a wide range of noncultivated diazotrophs. These noncultivated diazotrophs remain uncharacterized, as do their contributions to nitrogen fixation in natural systems. We have employed a novel 15N2-DNA stable isotope probing (5N2-DNA-SIP) method to identify free-living diazotrophs in soil that are responsible for nitrogen fixation in situ. Analyses of 16S rRNA genes from 15N-labeled DNA provide evidence for nitrogen fixation by three microbial groups, one of which belongs to the Rhizobiales while the other two represent deeply divergent lineages of noncultivated bacteria within the Betaproteobacteria and Actinobacteria, respectively. Analysis of nifH genes from 15N-labeled DNA also revealed three microbial groups, one of which was associated with Alphaproteobacteria while the others were associated with two noncultivated groups that are deeply divergent within nifH cluster I. These results reveal that noncultivated free-living diazotrophs can mediate nitrogen fixation in soils and that 15N2-DNA-SIP can be used to gain access to DNA from these organisms. In addition, this research provides the first evidence for nitrogen fixation by Actinobacteria outside of the order Actinomycetales.

Diversity of Planctomycetes in soil in relation to soil history and environmental heterogeneity.

Members of the Planctomycetes, which were once thought to occur primarily in aquatic environments, have been discovered in soils on five continents, revealing that these Bacteria are a widespread and numerically abundant component of microbial communities in soil. We examined the diversity of Planctomycetes in soil samples obtained from experimental plots at an agricultural site in order to assess the extent of Planctomycetes diversity in soil, to determine whether management effects such as past land cover and compost addition affected the composition of the Planctomycetes community, and to determine whether the observations made could provide insight into the ecological distribution of these organisms. Analysis of Planctomycetes 16S rRNA gene sequences revealed a total of 312 +/- 35 unique phylotypes in the soil at the site examined. The majority of these Planctomycetes sequences were unique, and the sequences had phylogenetic affiliations that included all major lineages in the Planctomycetaceae, as well as several novel groups of deeply divergent Planctomycetes. Both soil management history and compost amendment had significant effects on the Planctomycetes diversity, and variations in soil organic matter, Ca2+ content, and pH were associated with variations in the Planctomycetes community composition. In addition, Planctomycetes richness increased in proportion to the area sampled and was correlated with the spatial heterogeneity of nitrate, which was associated with the soil management history at the orchard site examined. This report provides the first systematic assessment of the diversity of Planctomycetes in soil and also provides evidence that the diversity of this group increases with area as defined by the general power law description of the taxon-area relationship.

A tissue-type plasminogen activator mutant with prolonged clearance in vivo. Effect of removal of the growth factor domain.

The complete cDNA for human tissue-type plasminogen activator (t-PA) was cloned and sequenced. A mutant was constructed by using in vitro site-specific mutagenesis to delete the region encoding the growth factor domain (amino acids 51-87 inclusive). Normal and mutant t-PA species were produced using two mammalian expression systems (in human HeLa cells and mouse C127 cells). The clearance of mutant and normal t-PA from plasma was examined in vivo using a guinea pig model. Mutant t-PA derived from HeLa or C127 cells was cleared much more slowly than the cognate normal t-PA. The potential role of the growth factor domain in the recognition of t-PA by the hepatic clearance mechanism is discussed.