Sequencing genomes from single cells by polymerase cloning.

Genome sequencing currently requires DNA from pools of numerous nearly identical cells (clones), leaving the genome sequences of many difficult-to-culture microorganisms unattainable. We report a sequencing strategy that eliminates culturing of microorganisms by using real-time isothermal amplification to form polymerase clones (plones) from the DNA of single cells. Two Escherichia coli plones, analyzed by Affymetrix chip hybridization, demonstrate that plonal amplification is specific and the bias is randomly distributed.

[Chronic post-mastectomy pain].

Background: Surgery results in chronic pain in 7-80 percent. One of the most studied is chronic post-mastectomy pain. The prevalence was 40-50 percent in studies performed abroad.

Identity, harm, and the ethics of reproductive technology.

The controversial question of whether a future child can be harmed by the use of reproductive technology turns on the way that the future child's identity is understood. As a result, analysis of the ethical and legal obligations to the children of reproductive technology that are based upon the possibility of such harm depends upon the conception of identity that is used. This paper reviews the contributions of two recent books, David DeGrazia's Human Identity and Bioethics (2005) and Philip Peters' How Safe is Safe Enough? (2004) to this area of inquiry.

[Patients' perioperative worries and experiences].

Aim: The aim of this study was to detect the main causes of patients' preoperative distress and their subjective experiences.

Methods: After institutional ethic committee approval and patients consent an anonymous questionnaire was randomly distributed on the first and second postoperative day. Its content were basic demography, preoperative anxiety level using five point verbal scale, main cause; of anxiety, the worst postoperative experience, postoperative pain intensity (11 point scale) and its treatment incidence of postoperative nausea and vomiting (PONV) and other side effects of the surgery and the anesthesia.

The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).

The National Institutes of Health's Mammalian Gene Collection (MGC) project was designed to generate and sequence a publicly accessible cDNA resource containing a complete open reading frame (ORF) for every human and mouse gene. The project initially used a random strategy to select clones from a large number of cDNA libraries from diverse tissues. Candidate clones were chosen based on 5'-EST sequences, and then fully sequenced to high accuracy and analyzed by algorithms developed for this project.

[Diclofenac 75mg. and 30 mg. orfenadine (Neodolpasse) versus placebo and piroxicam in postoperative analgesia after arthroscopy].

Purpose Of The Study: Arthroscopy is often performed in an out-patient department or as one-day surgery. Opioids often used as postoperative analgesics may have unwanted side effects that may postpone the patient's discharge from hospital. This study was designed to evaluate a substitute for the most frequently used opioid pethidine.

Comparison of the genome of the oral pathogen Treponema denticola with other spirochete genomes.

We present the complete 2,843,201-bp genome sequence of Treponema denticola (ATCC 35405) an oral spirochete associated with periodontal disease. Analysis of the T. denticola genome reveals factors mediating coaggregation, cell signaling, stress protection, and other competitive and cooperative measures, consistent with its pathogenic nature and lifestyle within the mixed-species environment of subgingival dental plaque.

Protein interaction mapping on a functional shotgun sequence of Rickettsia sibirica.

Protein interaction maps can reveal novel pathways and functional complexes, allowing 'guilt by association' annotation of uncharacterized proteins. To address the need for large-scale protein interaction analyses, a bacterial two-hybrid system was coupled with a whole genome shotgun sequencing approach for microbial genome analysis. We report the first large-scale proteomics study using this system, integrating de novo genome sequencing with functional interaction mapping and annotation in a high-throughput format.

Identifying genes for neuron survival and axon outgrowth in Hirudo medicinalis.

We have studied the molecular basis of nervous system repair in invertebrate (Hirudo medicinalis) nerve cells. Unlike in mammals, neurons in invertebrates survive injury and regrow processes to restore the connections that they held before the damage occurred. To identify genes whose expression is regulated after injury, we have used subtractive probes, constructed from regenerating and non-regenerating ganglia from the leech Hirudo medicinalis, to screen cDNA libraries made from whole leech CNS or from identified microdissected neurons.

Vascular endothelial growth factor in diabetic nephropathy.

Background: Vascular endothelial growth factor (VEGF) increases endothelial permeability. VEGF is produced in podocytes and functional receptors are located on endothelial glomerular cells. The aim of the current study in diabetic patients with normal renal function to various degrees of proteinuric nephropathy was therefore to unravel a possible role of the most important isoform VEGF(165) for albuminuria and to investigate the impact of therapy with an inhibitor of the renin-angiotensin system on VEGF(165) secretion.

Evidence for two types of low-energy charge transfer excitations in Sr2CuO3.

A comparative analysis of electron energy-loss spectroscopy (EELS) spectra for the 1D insulating cuprate Sr2CuO3 with transferred momentum q--> axially and radially to the chain axis allows one to elucidate the structure of the charge transfer gap in in-chain response. It is determined by the superposition of two types of excitations with different magnitudes of dispersion. The low-energy response with q--> radially to the chain direction, but yet within the plane of CuO4 plaquettes, exhibits also a dispersionless peak near 2 eV.

Single-cell analysis reveals cell-specific patterns of expression of a family of putative voltage-gated sodium channel genes in the leech.

To understand the molecular basis of nervous system function in the leech, Hirudo medicinalis, we have isolated four novel cDNAs encoding putative voltage-gated sodium (Na) channel alpha subunits, and have analyzed the expression of these genes in individual neurons of known function. To begin, degenerate oligonucleotide primers were used in combination with pre-existing cDNA libraries and reverse transcriptase-coupled polymerase chain reactions (RT-PCR). The putative leech Na channel cDNAs (LeNas) exhibit a higher degree of sequence homology to Na channel genes in other species than to voltage-gated calcium or potassium channel genes, including those expressed in leech.

[Progress in surgical treatment may improve the quality of life in patients with breast surgery for malignant tumors].

Breast cancer surgery (BCS) is a common procedure performed in women. Chronic postmastectomy pain (PMP) has been reported in as many as 22-72 percent of patients. There are no published reports in Czech Republic.

Metabolic effects of hypothermia and its neuroprotective effects on the recovery of metabolic and electrophysiological function in the ischemic retina in vitro.

Objective: Reduction in energy usage has been investigated as the mechanism by which hypothermia provides protection during ischemia. We describe experiments using hypothermia in the rabbit retina in vitro that show a correlation between hypothermia-induced reductions in energy usage and neuroprotection.

Methods: We examined energy metabolism and electrophysiological function under control/nonischemic conditions during 1 or 2 hours of "ischemia" (induced by decreasing glucose from 6 to 1 mmol/L and oxygen from 95 to 15%) and during 3 to 4 hours of "return-to-control" conditions.

Radiation hybrid map, physical map, and low-pass genomic sequence of the canine prcd region on CFA9 and comparative mapping with the syntenic region on human chromosome 17.

Progressive rod-cone degeneration (prcd) is a canine retinal disease that maps to the centromeric end of CFA9 in a region of synteny with the distal part of HSA17q. As such, prcd has been postulated as the only animal model of RP17, a human retinitis pigmentosa locus that maps to 17q22. In an effort to establish more detailed regions of synteny between dog CFA9 and the HSA17q-ter region, we created a robust gene-enriched CFA9-RH08(3000) map with 34 gene-based markers and 12 microsatellites, with the highest resolution and number of markers for the centromeric end of CFA9.

[Infiltration of local anesthetics into the thyroid gland capsule for surgery and the postoperative period].

In a prospective randomized study the hypothesis was tested whether infiltration of the thyroid capsule by a local anaesthetic will reduce the haemodynamic response to surgical trauma, consumption of anaesthetics and opioids during surgery and will shorten the time of arousal. A total of 64 patients indicated for planned goitre surgery were divided at random into a control group (C, n = 32) and experimental group (LA, n = 32). The preoperative medication and anesthesia did not differ in the two groups.

Combined microsurgical and endovascular management of complex intracranial aneurysms.

Objective: The disciplines of microneurosurgery and cranial base surgery have reached maturity, and technical advances in the surgical management of aneurysms are limited. Although most aneurysms can be clipped microsurgically or coiled endovascularly, a subset of patients may require a combined approach. A consecutive series of patients with aneurysms in one surgeon's cerebrovascular practice was reviewed retrospectively to analyze strategies for integrating microsurgical and endovascular techniques in the management of complex aneurysms.

New type of anaesthesia with sevofluran improves surgical and post-surgical course of burns treatment (IGA 6116/2).

The objectives of this open-perspective clinical study were to test the effect of a new type of anaesthesia using sevofluran during surgical treatment of patients with burns, and to compare it with that of ketamin, at present the most frequently used anaesthetic. The study, conceived as a pilot study, was performed on 10 paediatric and 8 adult patients with 2nd degree burns covering between 7% and 35% of body surface. Owing to highly significant differences in parameters in favour of sevofluran, it was recommended by our ethical committee not to increase the number of patients.

Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.

The National Institutes of Health Mammalian Gene Collection (MGC) Program is a multiinstitutional effort to identify and sequence a cDNA clone containing a complete ORF for each human and mouse gene. ESTs were generated from libraries enriched for full-length cDNAs and analyzed to identify candidate full-ORF clones, which then were sequenced to high accuracy. The MGC has currently sequenced and verified the full ORF for a nonredundant set of >9,000 human and >6,000 mouse genes.

The genome sequence of the malaria mosquito Anopheles gambiae.

Anopheles gambiae is the principal vector of malaria, a disease that afflicts more than 500 million people and causes more than 1 million deaths each year. Tenfold shotgun sequence coverage was obtained from the PEST strain of A. gambiae and assembled into scaffolds that span 278 million base pairs.

The Brucella suis genome reveals fundamental similarities between animal and plant pathogens and symbionts.

The 3.31-Mb genome sequence of the intracellular pathogen and potential bioterrorism agent, Brucella suis, was determined. Comparison of B.

A physical map of the mouse genome.

A physical map of a genome is an essential guide for navigation, allowing the location of any gene or other landmark in the chromosomal DNA. We have constructed a physical map of the mouse genome that contains 296 contigs of overlapping bacterial clones and 16,992 unique markers. The mouse contigs were aligned to the human genome sequence on the basis of 51,486 homology matches, thus enabling use of the conserved synteny (correspondence between chromosome blocks) of the two genomes to accelerate construction of the mouse map.

Effect of agroclavine on NK activity in vivo under normal and stress conditions in rats.

Agroclavine is a natural, clavine type of ergot alkaloid with D1 dopamine and a-adrenoceptor agonistic properties. We showed previously that in vitro agroclavine enhances natural killer (NK) cell activity, increases interleukin-2 and interferon-gamma production and prolongs the survival time of tumor-bearing mice. The aim of this study was 1) to test the effect of agroclavine on NK activity in vivo, and 2) to assess the potential toxicity of high doses of agroclavine on cardiac and liver functions using creatine kinase MB (CKMB) and alanine aminotransferase (ALT) as biochemical markers in normal and stressed animals.