Erratum to: Development of novel multiplex microsatellite polymerase chain reactions to enable high-throughput population genetic studies of Schistosoma haematobium.

Unfortunately, the original version of this article [1], contained a mistake. In Table 1, the primers for Sh6 and Sh9 were included incorrectly. Instead of GGGATGTATGCAGACTTG TTGTTTGGCTGCAGTAAC and GCTGAGCTTGAGATTG CTTCTGTCCCATCGATACC they should have been Sh6 Forward Primer GGTGGATTACGCAATAG, Sh6 Reverse Primer TTTAATCAACCGGGTGTC and Sh9 Forward Primer GGGATGTATGCAGACTTG, Sh9 Reverse Primer TTGTTTGGCTGCAGTAAC respectively. A corrected version of Table 1 is included below

Development of novel multiplex microsatellite polymerase chain reactions to enable high-throughput population genetic studies of Schistosoma haematobium.

BACKGROUND: Human urogenital schistosomiasis caused by Schistosoma haematobium is widely distributed across Africa and is increasingly targeted for control and regional elimination. The development of new high-throughput, cost-effective molecular tools and approaches are needed to monitor and evaluate the impact of control programs on the parasite populations. Microsatellite loci are genetic markers that can be used to investigate how parasite populations change over time and in relation to external influences such as control interventions. FINDINGS: Here, 18 existing S. haematobium microsatellite loci were optimised to enable simultaneous amplification across two novel multiplex microsatellite PCR's, each containing nine loci. Methods were developed for the cost effective and rapid processing and microsatellite analysis of S. haematobium larval stages stored on Whatman-FTA cards and proved robust on miracidia and cercariae collected from Zanzibar and Niger. CONCLUSION: The development of these novel and robust multiplex microsatellite assays, in combination with an improved protocol to elute gDNA from Whatman-FTA fixed schistosome larval stages, enables the high-throughput population genetic analysis of S. haematobium. The molecular resources and protocols described here advance the way researchers can perform multi locus-based population genetic analyses of S. haematobium as part of the evaluation and monitoring of schistosomiasis control programmes.

Population genetics of Schistosoma haematobium: development of novel microsatellite markers and their application to schistosomiasis control in Mali.

The recent implementation of mass drug administration (MDA) for control of uro-genital schistosomiasis has identified an urgent need for molecular markers to both directly monitor the impact of MDA, for example to distinguish re-infections from uncleared infections, as well as understand aspects of parasite reproduction and gene flow which might predict evolutionary change, such as the development and spread of drug resistance. We report the development of a novel microsatellite tool-kit allowing, for the first time, robust genetic analysis of individual S. haematobium larvae collected directly from infected human hosts. We genotyped the parasite populations of 47 children from 2 schools in the Ségou region of Mali, the first microsatellite study of this highly neglected parasite. There was only limited evidence of population subdivision between individual children or between the two schools, suggesting that few barriers to gene flow exist in this population. Complex relationships between parasite reproductive success, infection intensity and host age and gender were identified. Older children and boys harboured more diverse infections, as measured by the number of unique adult genotypes present. Individual parasite genotypes had variable reproductive success both across hosts, a pre-requisite for evolutionary selection, and, phenotypically, in hosts of different ages and genders. These data serve as a baseline against which to measure the effect of treatment on parasite population genetics in this region of Mali, and the tools developed are suitable to further investigate this important pathogen, and its close relatives, throughout their range.

Isolation and characterization of the first polymorphic microsatellite markers for Schistosoma haematobium and their application in multiplex reactions of larval stages.

The ability of microsatellite loci to reveal genetic diversity within the trematode Schistosoma haematobium is demonstrated for the first time. Nine novel polymorphic microsatellite markers were isolated and their viability assessed on 36 S. haematobium adult worm individuals from three geographical populations. Allelic diversity and gene diversity ranged from two to seven and from 0.29 to 0.76, respectively, suggesting high variability between individuals and between unrelated populations. Three primers also amplified Schistosoma mansoni and two Schistosoma japonicum. The results suggest these primers are useful for population genetic analyses of S. haematobium.

Assignment of paternity groups without access to parental genotypes: multiple mating and developmental plasticity in squid.

We present a novel approach to investigating sibling relationships and reconstructing parental genotypes from a progeny array. The Bayesian method we have employed is flexible and may be applicable to a variety of situations in addition to the one presented here. While mutation rates and breeding population allele frequencies can be taken into account, the model requires relatively few loci and makes few assumptions. Paternity of 270 veined squid (Loligo forbesi) hatchlings from three egg strings collected from one location was assigned using five microsatellite loci. Paternal and maternal genotypes reconstructed for each of the three strings were identical, strongly indicating that a single female produced the strings that were fertilized by the same four males. The proportion of eggs fertilized was not equal between males in all three strings, with male 1 siring most offspring (up to 68% in string 1), through to male 4 siring the least (as low as 2.4% in string 1). Although temperature had a profound effect on incubation time, paternity did not affect this trait at 12 degrees C or 8 degrees C.

A case for minimally invasive coronary surgery as primary treatment for left anterior descending coronary artery disease.

OBJECTIVE: The introduction of minimally invasive coronary bypass surgery has allowed the application of multiple approaches to coronary artery disease. METHODS: Technologic developments have resolved patency and myocardial ischemic issues and increased surgical experience and training have combined to make more coronary bypass surgeons facile in minimally invasive surgical techniques. RESULTS: These advances, along with the decreased invasiveness, shortened recovery and lower cost, suggest the application of these techniques to the primary treatment of disease of the anterior descending artery. CONCLUSION: In selected circumstances such as these in which patient co-morbid risk factors would suggest high recurrence rates in complex lesions or total occlusion of the anterior descending coronary artery and present in cases of in-stent restenosis, primary application of minimally invasive bypass techniques may create long-term benefits for patients.

Clinical evaluation of the on-line Sensicath blood gas monitoring system.

BACKGROUND: Blood gas analysis is an integral part of the management of open heart surgery and post-surgical intensive care patients. This study was conducted to compare the results of on-line blood gas analysis using the new Sensicath (SC) optical sensor technology against standard blood gas assay. METHODS: Comparative blood gas analysis was performed on 45 postoperative cardiovascular surgical patients and 5 patients during cardiopulmonary bypass. Simultaneous samples were obtained. One sample was sent to the central hospital laboratory for measurement using a Radiometer ABL 500 blood gas analyzer and the second sample was drawn past the optical sensor. Comparisons between the two methods for pH, pO2 and pCO2 were analyzed using linear regression and the method of Bland and Altman for the intensive care unit samples (N = 451) while the T test was used to compare data points obtained during cardiopulmonary bypass (N = 57). Bias, accuracy and precision were also calculated. RESULTS: The regression lines (r2) for pH, pO2, and pCO2 and were 0.69, 0.94, and 0.68 with slopes of 1.000, 0.0957, and 1.052 respectively for postoperative surgical patients. During cardiopulmonary bypass, the T test did not show any significant difference between the Sensicath and the reference values for the arterial and venous pH, pO2 and pCO2. CONCLUSIONS: In summary, this study found the Sensicath to be an accurate, useful tool which permits immediate blood gas analysis without blood exposure and constitutes an advance in the field of intensive care monitoring.

Insect Na(+)/K(+)-ATPase.

Na(+)/K(+)-ATPase (sodium/potassium pump) is a P-type ion-motive ATPase found in the plasma membranes of animal cels. In vertebrates, the functions of this enzyme in nerves, heart and kidney are well characterized and characteristics a defined by different isoforms. In contrast, despite different tissue distributions, insects possess a single isoform of the alpha-subunit. A comparison of insect and vertebrate Na(+)/K(+)-ATPases reveals that although the mode of action and structure are very highly conserved, the specific roles of the enzyme in most tissues varies. However, the enzyme is essential for the function of nerve cells, and in this respect Na(+)/K(+)-ATPase appears to be fundamental in metazoan evolution.

Minimally invasive direct coronary artery bypass grafting: experimental and clinical experiences.

BACKGROUND: This communication briefly details the goals, indications, surgical approaches, and limitations of minimally invasive direct coronary artery bypass grafting (MIDCABG). The experimental experiences from various institutions are summarized. METHODS: The clinical experiences of 72 consecutive MIDCABG procedures performed at our institutions between June 5, 1995, and August 13, 1996, were analyzed. We have divided patients into two groups. Group A consists of healthy low-risk patients with single lesions of the left anterior descending coronary artery or the right coronary artery, or with both lesions of both arteries. Group B consists of high-risk patients who had major contraindications to conventional cardiopulmonary bypass procedures. There were 55 patients in group A and 17 patients in group B. Using The Society of Thoracic Surgeons preoperative predicted risk module, group A had a 1% predicted mortality versus 4% in group B. RESULTS: The 30-day mortality was 2% in group A and 6% in group B. The mean postoperative length of stay was 4 days for group A and 5 1/2 days for group B. Short-term follow-up of the survivors appears promising, and 81% of patients were angina free at the time of last follow-up. CONCLUSIONS: The MIDCABG techniques are still developing. The short-term results during the learning period appear to be quite good, but long-term results remain yet to be seen. The addition of new equipment to facilitate construction of the anastomosis will enhance application and results. The lessons learned from these approaches are already being applied to other areas of cardiac surgery including valve replacement and the repair of congenital heart defects.

Revascularization using angioplasty and minimally invasive techniques documented by thermal imaging.

Minimally invasive direct coronary artery bypass grafting offers mortality and morbidity advantages to selected patients. To broaden indications for such, an appropriate and combined disciplinary approach using angioplasty and minimally invasive direct coronary artery bypass grafting is described in a patient requiring reoperative grafting. Documentation of patency of new left internal mammary artery-to-left anterior descending artery anastomoses performed without the use of cardiopulmonary bypass was obtained intraoperatively using a Thermal Imaging Camera.

A single isoform of the Na+/K(+)-ATPase alpha-subunit in Diptera: evidence from characterization of the first extracellular domain.

The first extracellular domain of the alpha-subunit of the Na+/K+-ATPase (sodium/potassium pump) is functionally important, affecting sensitivity of the enzyme to cardiac glycosides (e.g. ouabain) and being implicated in the transport of K+. This domain is also variable among mammalian isoforms of the alpha-subunit. Using PCR, we have isolated from seven insect species with contrasting physiologies a DNA fragment containing this region, in order to help determine whether tissue-specific expression might be associated with isoforms encoded by a gene family, as it is in mammals. A single sequence (with one ORF) characteristic of Na+/K+-ATPase was obtained from genomic DNA of each species. Only the fragment from Manduca sexta contained an intron, but at a location different to that found in mammals. For all Diptera so far characterized, the species phylogeny is the same as the alpha-subunit gene phylogeny (based on the sequences of the first extracellular domain and flanking transmembrane domains). The results strongly indicate a single, ouabain-sensitive isoform of the alpha-subunit of Na+/K+-ATPase is present in Diptera.

The Vale of Leven health promotion project.

A two stage health promotion programme is in progress in the Vale of Leven in Dunbartonshire. The first stage has been completed within a local factory (Polaroid UK Limited), the largest private employer in the district. A total of 1205 employees, representing 87% of the workforce, took part in the programme and were initially screened for coronary artery risk factors. Blood pressure, serum cholesterol, body composition and aerobic fitness were measured and smoking habits determined. Aspects of lifestyle were assessed by questionnaire. All employees whose initial cholesterol concentration was greater than 6.5 mM were given simple dietary advice and their cholesterol concentration thereafter remeasured. Eighty-two per cent of these men and 72% of these women succeeded in reducing their cholesterol, the men by a mean of 1.3 mM, the women by a mean of 0.7 mM. The health initiatives undertaken within the factory at the same time as screening are also described in this paper.