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PubMed1990

The ATPase activity of SecA is regulated by acidic phospholipids, SecY, and the leader and mature domains of precursor proteins.

The ATPase activity of SecA is stimulated by E. coli plasma membrane vesicles bearing SecY protein and a precursor protein such as proOmpA. This activity is termed "translocation ATPase". Liposomes alone can also stimulate SecA ATPase, but membrane proteins block this stimulation in native inner membranes. We define the stimulation of SecA ATPase by lipid as "SecA/lipid ATPase". SecA/lipid ATPase, translocation ATPase, and translocation into inner membrane vesicles require acidic phospholipids, suggesting an underlying unity of mechanism. ProOmpA and ATP stabilize liposome-bound SecA. Full SecA/lipid ATPase activity and stability are also seen when a mixture of a leader peptide and either OmpA or maltose binding protein (MBP) are added instead of proOmpA, while neither the leader peptide alone nor OmpA or MBP suffice. Cytosolic proteins in conjuction with a leader peptide are less active in this reaction, indicating that liposome-bound SecA protein recognizes both leader and mature domains.

Adenosine Triphosphatases↗

PubMed1976

Acute effects of oral and intravenous ethanol on rat hepatic enzyme activities.

1. Oral administration of ethanol (3 ml) of 95% in 12 ml total volume over a two day period) significantly decrease plasma glucose and insulin levels and the activities of two key gluconeogenic enzymes, pyruvate carboxylase (pyruvate: CO2 ligase (ADP), EC 6.4.1.1) and fructose diphosphatase, (D-Fru-1,6-P2 1-phosphohydrolase, EC 3.1.3.11), and one glycolytic enzyme, fructose-1,6-P2 aldolase (Fru-1,6-P2 D-glyceraldehyde-3-P lyase, EC 4.1.2.13). In each instance, the administration of 2400 mug daily of oral folate in conjuction with the ethanol prevented these alterations in carbohydrate metabolism. 2. Intravenous injection of ethanol produced a rapid decrease (within 10--15 min) in the activities of hepatic phosphofructokinase, (ATP:D-fructose-6-phosphate 6-phosphotransferase, EC 2.7.1.11), pyruvate kinase, (ATP:pyruvate phosphotransferase, EC 2.7.1.40), fructose diphosphatase and fructose-1,6-P2 aldolase. 3. Intravenous ethanol significantly increased hepatic cyclic AMP concentration approximately 60% within 10 min, while oral ethanol did not alter hepatic cyclic AMP concentrations. 4. These data confirm the known antagonism ethanol and folate and suggest that oral folate might offer a protective effect against hypoglycemia in rats receiving ethanol.

Administration, Oral↗

PubMed1979

Automated scene analysis of CT scans.

Since the advent of computed tomography, there has been an increasing realization that CT scans contain quantitative as well as qualitative information useful in the diagnostic process. Often however, the use of this information is impeded by the tedious manual outlining of the areas of interest in the scan. To alleviate this problem, we have developed a scene segmentation algorithm which will automatically delineate areas of interest in a CT scan. This procedure uses known information about the expected objects in the scan in conjuction with an algorithm to label those objects. The resultant scene segmentation divides the scan into four anatomical areas: skull, normal brain, high density lesions and CSF. After an area of interest is interactively selected by the clinician, volume, density or other quantitative measures may be computed. Limitations of the algorithm and its clinical applications are discussed.

Absorptiometry, Photon↗

PubMed1979

Inceptor and origin of DNA replication in lambdoid coliphages. II. The lambda DNA maximal replication system.

In pBR313-lambda dv hydrid plasmids a second system for initiation of DNA replication has been detected in lambdoid replicator DNAs (in the absence of the p0 promoter). The "maximal" (or "maxi") initiation system depends on the origin of replication (ori) sequence, in conjuction with the "inceptor" (ice) element located in the lambdoid cII genes. Only leftward, but not bidirectional, primer RNA synthesis seems to be initiated at ori in its newly defined boundaries, and it appears to be catalysed by dnaG-coded primase. Only if transcriptionally activated, will ori effectively initiate lambda specific, O and P-dependent "maximal" hybrid-plasmid replication. In addition, it will repress a complete lambda "minimal" initiation system in cis, i.e., if present on the same plasmid molecule. This newly discovered repressive activity of the ori system depends on only three factors: an intact left section of ori, the O product, and transcriptional activation of ori (rightward or leftward). A repressed minimal initiation system will regain its activity as soon as a segment carrying either part of the O gene or a promoter for transcriptional activation is delected from such a plasmid which was combining both the "mini" and "maxi" systems of lambda replication.

Bacteriophage lambda↗

PubMed1993

Rapid identification of proteins by peptide-mass fingerprinting.

BACKGROUND: Developments in 'soft' ionisation techniques have revolutionized mass-spectro-metric approaches for the analysis of protein structure. For more than a decade, such techniques have been used, in conjuction with digestion b specific proteases, to produce accurate peptide molecular weight 'fingerprints' of proteins. These fingerprints have commonly been used to screen known proteins, in order to detect errors of translation, to characterize post-translational modifications and to assign diulphide bonds. However, the extent to which peptide-mass information can be used alone to identify unknown sample proteins, independent of other analytical methods such as protein sequence analysis, has remained largely unexplored. RESULTS: We report here on the development of the molecular weight search (MOWSE) peptide-mass database at the SERC Daresbury Laboratory. Practical experience has shown that sample proteins can be uniquely identified from a few as three or four experimentally determined peptide masses when these are screened against a fragment database that is derived from over 50 000 proteins. Experimental errors of a few Daltons are tolerated by the scoring algorithms, thus permitting the use of inexpensive time-of-flight mass spectrometers. As with other types of physical data, such as amino-acid composition or linear sequence, peptide masses provide a set of determinants that are sufficiently discriminating to identify or match unknown sample proteins. CONCLUSION: Peptide-mass fingerprints can prove as discriminating as linear peptide sequences, but can be obtained in a fraction of the time using less protein. In many cases, this allows for a rapid identification of a sample protein before committing it to protein sequence analysis. Fragment masses also provide information, at the protein level, that is complementary to the information provided by large-scale DNA sequencing or mapping projects.

Journal Article↗

PubMed2005

Structure-activity studies of new melanocortin peptides containing an aromatic amino acid at the N-terminal position.

Cyclic melanotropin peptides, designed with an aromatic amino acid substitution at the N-terminal position of the MT-II-type scaffold, were prepared by solid-phase peptide synthesis and evaluated for their ability to bind to and activate human melanocortin-1, -3, -4, and -5 receptors. The structure-activity studies of these MT-II analogues have identified a selective antagonist at the hMC4R (H-Phe-c[Asp-Pro-d-Nal(2')-Arg-Trp-Gly-Lys]-NH(2), pA(2)=8.7), a selective partial agonist at the hMC4R (H-d-Nal(2')-c[Asp-Pro-d-Phe-Arg-Trp-Gly-Lys]-NH(2), IC(50)=11nM, EC(50)=56nM), and a selective partial agonist at the hMC3R (H-d-Phe-c[Asp-Pro-d-Phe-Arg-Trp-Lys]-NH(2), IC(50)=3.7nM, EC(50)=4.9nM). Aromatic amino acid substitution at the N-terminus in conjuction with the expansion of the 23-membered cyclic lactam MT-II scaffold to a 26-membered scaffold by addition of a Gly residue in position 10 leads to melanotropin peptides with enhanced receptor selectivity.

Amino Acid Sequence↗

PubMed2005

The cost of steroid-free protocols in primary cadaver kidney transplantation.

Steroid-free immunosuppression regimens have been enjoying recent success in clinical transplantation. The use of antibodies required for such protocols can be an economic burden. We proposed to study their cost in our center. This retrospective study involved 147 consecutive patients subjected to 4 protocols of immunosuppression. The first received triple therapy. The second group received induction with basiliximab, whereas the third received Basiliximab plus cyclosporine (CSA) plus mycophenolate mofetil (MMF), and the fourth received Thymoglobulin plus CSA plus MMF in conjuction with only 4 days of steroid. Rejection episodes were treated with Solumedrol. Six-month charges were obtained from computerized records of the finance department, the in-house laboratories, and the transplantation service registry. All charges were expressed in 2004 dollars. Statistical analyses were obtained using chi-square, analysis of variance (ANOVA) and Kaplan-Meier tests. The 4 groups were similar with regard to donor and/or recipient gender, race, panel reactive antibodies, cold ischemia, dialysis requirements length of stay and readmission, graft survival, and function. Charges were significantly higher in the last 2 groups as compared with triple therapy.

Adult↗

PubMed1977

Camera-computer systems for rapid dynamic imaging studies.

The extraction of valuable diagnostic information from rapid, dynamic studies performed on a gamma camera requires that the data be recorded for latter analysis. If these data are to be available rapidly and if quantitative analysis is to be performed, it is imperative that a computer be used in conjuction with the gamma camera. This review describes the more important features of a nuclear medicine computer system both from the point of view of hardware and software. The various aspects that cause some limitations to be imposed on the capability of the system in order to carry out fast, dynamic studies are described and some possible solutions are suggested.

Analog-Digital Conversion↗

PubMed1977

Osmotic correction to elastic area compressibility measurements on red cell membrane.

In a recent article (Biophys. J. 16:585, 1976), we reported measurements of the elastic area compressibility modulus or red cell membranes using micropipette aspiration on osmotically preswollen red cells. Subsequently, we have analyzed the effects of osmotic and hydrostatic pressure driving forces across the cell membrane in conjuction with the mass conservation equation; we find that the change in cell volume due to the reversible movement of water out of the cell can produce one-third of the movement of the cell projection in the pipette tip. Since the actual volume changes is too small to measure directly (about 1% of the total cell volume), we have used an indirect experimental method to provide critical evaluation of the analysis of cell volume change versus applied pressure; this is based on the model that the change in cell volume is inversely proportional to the cellular osmotic strength. We have increased the cellular cation concentration with a drug, nystatin, and measured the elastic area compressibility modulus corrected for osmotic volume changes as a function of cellular osmotic strength. We find that the corrected elastic are compressibility modulus is independent of cellular osmotic strength, which supports the model and calculated correction for the osmotic effect. The elastic area compressibility modulus is 450 dyn/cm at 25 degrees C instead of 300 dyn/cm, determined previously.

Biological Transport↗

PubMed1997

Control of phobic behavioral responses by rhodopsin-induced photocurrents in Chlamydomonas.

Both phototactic and photophobic responses of Chlamydomonas are mediated by a visual system comprising a rhodopsin photoreceptor. Suction pipette recordings have revealed that flash stimulation causes calcium currents into the eyespot and the flagella. These photocurrents have been suggested to be the trigger for all behavioral light responses of the cell. But this has never been shown experimentally. Here we describe a detection technique that combines electrical and optical measurements from individual algae held in a suction pipette. Thus it is possible to record photocurrents and flagellar beating simultaneously and establish a direct link between the two. We demonstrate that in Chlamydomonas only the photoreceptor current in conjuction with a fast flagellar current constitutes the trigger for photophobic responses. Within the time of the action-potential-like flagellar current, the flagella switch from forward to backward swimming, which constitutes the beginning of the photoshock reaction. The switch is accompanied by a complex frequency change and beating pattern modulation. The results are interpreted in terms of a general model for phototransduction in green algae (Chlorophyceae).

Animals↗

PubMed1977

A nitrosamide-specific detector for use with high-pressure liquid chromatography.

A highly specific automated method for identification of nitrosamides has been developed for use in conjuction with reversed-phase high-pressure liquid chromatography. The method is based on the well-known Griess reagent, and is capable of sensitivities of 0.5 nmoles of injected nitrosamide in most cases. The method may be used for analysis of nitrite, nitrosamides, nitrosocarbamates, and alkylnitrites.

Alkylation↗

PubMed1977

Serum iron concentration and transferrin saturation in the diagnosis of iron deficiency in children: normal developmental changes.

A group of 359 healthy children and 49 adults were studied for the purpose of estimating the normal limits for serum iron concentration and transferrin saturation. The 144 children and seven adults who has any other laboratory evidence of iron deficiency (abnormal values of serum ferritin, free erythrocyte protoporphyrin, hemoglobin concentration, or mean corpuscular volume) were excluded. In evaluating the 215 children and 42 adults who met the criteria to be considered normal we found that serum iron concentration and transferrin saturation were significantly lower in children between the ages of 0.5 and 12 years than in adults. We conclude that in children between the ages of 0.5 and 12 years, a transferrin saturation of less than 16% constitutes good evidence of iron deficiency only in conjuction with anemia and low mean corpuscular volume.

Adolescent↗

PubMed1975

Post-obstructive diuresis.

Massive diuresis after relief of urinary tract obstruction has been divided into 3 categories: 1) salt, 2) urea and 3) water. Measurements of the urinary electrolytes and urine osmolality establish the character of the diuresis and facilitate in the fluid management of these patients. A clinical example of the 2 most common varieties, salt and urea, is presented in conjuction with the method of fluid replacement.

Diabetes Insipidus↗

PubMed2001

A pilot survey of blood lead levels in various types of workers in the United Arab Emirates.

The aim of the present study is to determine the effect of blood lead on the health of industrial workers in United Arab Emirates (UAE). This is a cross-sectional pilot survey of blood lead levels (BLL) of 100 industrial workers (exposed) and 100 non-industrial workers (nonexposed), matched for age, sex and nationality selected from Al-Ain, Abu-Dhabi Emirate. Industrial workers had significantly higher mean of BLL (77.5+/-42.8 microg/dl and median 80.9 microg/dl) than non-industrial workers (19.8+/-12.3 microg/dl and median 11.0 microg/dl). In the present study, reported symptoms among industrial workers were strongly associated with BLL nausea/vomiting, muscular symptoms, dizziness, fatigue, irritability, memory disturbances, insomnia and allergic conjuctivitis, rhinitis and dermatitis. Furthermore, the present study revealed that industrial workers had higher prevalence of respiratory symptoms for phlegm, shortness of breath and diagnosed asthma. In conclusion, this study determined that occurrence of certain symptoms might be associated with lead exposure among industrial workers.

Cross-Sectional Studies↗

PubMed1998

Detection of adenovirus outbreak at a municipal swimming pool by nested PCR amplification.

In July 1995 an outbreak of pharyngoconjuctivitis caused by adenoviruses occurred among athletes participating in a swimming contest in a town in southern Greece (Peloponnese). At least 80 persons displayed symptoms of the illness, with the predominant ones being high fever, sore throat, conjuctivitis, headache, and abdominal pain. Poor chlorination was probably the cause of the outbreak (residual chlorine <0.2 mg/l), as after hyperchlorination the spread of adenoviruses stopped. Rapid detection of adenoviruses in the municipal swimming pool water by nested polymerase chain reaction (PCR) amplification allowed quick control of the outbreak.

Adenoviridae↗

PubMed2003

[Osseous lithiasis: an unusual condition].

Bone lithiasis is an uncommon disease unfrequent reported in the literature. Its origin is supported on urothelium and conjuctive tissue bone metaplasia. High suspicious index is needed to establish its diagnosis; where radiological signs and extracorporeal lithotripsy resistence drive to these uncommon entity. Definitive diagnosis is based on histological exam. We report a 49-year-old patient with left obstructive uropathy due to a high ureteral lumbar lithiasis who precised ureterolithectomy and partial ureterectomy after extracorporeal litotripsy failure. Histological exam confirmed bone lithiasis and squamous metaplasia. Epidemiologic, diagnostic and therapeutic aspects are briefly reviewed in literature. Bone calculus treatment includes lithectomy and urothelium resection to avoid posterior recurrence. Possible association to tumoral transformation suggests close evolutive follow-up.

Humans↗

PubMed1998

Determination of nickel in saliva by electrothermal atomic absorption spectrometry using various chemical modifiers with Zeeman-effect background correction.

The profile of nickel signal using electrothermal atomic absorption spectrometry with deuterium and Zeeman-effect background correction is presented. The Zeeman effect system of background correction offered definitive advantages and therefore was used for the determination of nickel in saliva in the presence of various isomorphous metals. The highest nickel absorbance values corresponded at 200, 300, 300, 300, 600, and 200 ng of Tb, Mg, Sm, Lu, Tm, and Pd, respectively. On the other hand, the addition of Eu, Er, and Ho decreased the nickel signal. The presence on each modifier alone does not eliminate the matrix interference. However, the use of 200 ng of Pd in conjuction with 300 ng of Lu has a higher sensitivity, offers an advantage against interference from the background of saliva matrix and produces good recoveries (98 to 102% from unspiked and spiked saliva samples). The limit of detection was 0.11 micrograms/L for a characteristic mass of 16.6 pg of nickel using Pd-Lu as modifier. The within-batch precision varied between 0.8 and 1.5% relative standard deviations. The analysis of thirty samples of whole saliva gave an average of 0.81 +/- 0.30 of micrograms/L of Ni (range from 0.5 to 2.0 micrograms/L of Ni). The agreement between the observed and certified values obtained from a Seronorm Blood Serum Standard Reference Material was good.

Adult↗

PubMed2000

Value of pedigree/clinical data, immunohistochemistry and microsatellite instability analyses in reducing the cost of determining hMLH1 and hMSH2 gene mutations in patients with colorectal cancer.

The aim of this study was to evaluate the significance of pedigree/clinical data, immunohistochemistry (IHC) and microsatellite instability (MI) analyses in the reduction of costs of constitutional hMLH1 and hMSH2 gene mutation diagnosis in patients with colorectal cancers (CRC). Pedigree/clinical data were evaluated on a series of 168 patients with CRC, including 43 consecutive sporadic late-onset and 25 consecutive, definitive or suspected hereditary non-polyposis colorectal cancer (HNPCC) cases, examined by IHC and MI analyses. In the latter group, 6/25 (24%) constitutional mutations were found. We detected no germline mutations in the sporadic late-onset patients. The lowest costs (880 Euro/mutation detected) were achieved by performing pedigree/clinical data (for exclusion of late-onset sporadic CRC) in conjuction with IHC only. In this model 1/6 (17%) mutations was missed. Additional preselection by IHC and MI analyses before sequencing was required to detect all mutations. In this approach, which seems to be the most effective in the search for hMLH1 and hMSH2 gene mutation, the cost was 1767 euro/mutation detected.

Adaptor Proteins, Signal Transducing↗