ABRF ESRG 2006 study: Edman sequencing as a method for polypeptide quantitation.

The Edman Sequencing Research Group (ESRG) designs studies on the use of Edman degradation for protein and peptide analysis. These studies provide a means for participating laboratories to compare their analyses against a benchmark of those from other laboratories that provide this valuable service. The main purpose of the 2006 study was to determine how accurate Edman sequencing is for quantitative analysis of polypeptides. Secondarily, participants were asked to identify a modified amino acid residue, N-epsilon-acetyl lysine [Lys(Ac)], present within one of the peptides. The ESRG 2006 peptide mixture consisted of three synthetic peptides. The Peptide Standards Research Group (PSRG) provided two peptides, with the following sequences: KAQYARSVLLEKDAEPDILELATGYR (peptide B), and RQAKVLLYSGR (peptide C). The third peptide, peptide C*, synthesized and characterized by ESRG, was identical to peptide C but with acetyl lysine in position 4. The mixture consisted of 20% peptide B and 40% each of peptide C and its acetylated form, peptide C*. Participating laboratories were provided with two tubes, each containing 100 picomoles of the peptide mixture (as determined by quantitative amino acid analysis) and were asked to provide amino acid assignments, peak areas, retention times at each cycle, as well as initial and repetitive yield estimates for each peptide in the mixture. Details about instruments and parameters used in the analysis were also collected. Participants in the study with access to a mass spectrometer (MALDI-TOF or ESI) were asked to provide information about the relative peak areas of the peptides in the mixture as a comparison with the peptide quantitation results from Edman sequencing. Positive amino acid assignments were 88% correct for peptide C and 93% correct for peptide B. The absolute initial sequencing yields were an average of 67% for peptide (C+C*) and 65.6 % for peptide B. The relative molar ratios determined by Edman sequencing were an average of 4.27 (expected ratio of 4) for peptides (C+C*)/B, and 1.49 for peptide C*/C (expected ratio of 1); the seemingly high 49% error in quantification of Lys(Ac) in peptide C* can be attributed to commercial unavailability of its PTH standard. These values compare very favorably with the values obtained by mass spectrometry.

ABRF ESRG 2005 study: identification of seven modified amino acids by Edman sequencing.

Identification of modified amino acids can be a challenging part for Edman degradation sequence analysis, largely because they are not included among the commonly used phenylthiohydantion amino acid standards. Yet many can have unique retention times and can be assigned by an experienced researcher or through the use of a guide showing their typical chromatography characteristics. The Edman Sequencing Research Group (ESRG) 2005 study is a continuation of the 2004 study, in which the participating laboratories were provided a synthetic peptide and asked to identify the modified amino acids present in the sequence. The study sample provided an opportunity to sequence a peptide containing a variety of modified amino acids and note their retention times relative to the common amino acids. It also allowed the ESRG to compile the chromatographic properties and intensities from multiple instruments and tabulate an average elution position for these modified amino acids on commonly used instruments. Participating laboratories were given 2000 pmoles of a synthetic peptide, 18 amino acids long, containing the following modified amino acids: dimethyl- and trimethyl-lysine, 3-methyl-histidine, N-carbamyl-lysine, cystine, N-methyl-alanine, and isoaspartic acid. The modified amino acids were interspersed with standard amino acids to help in the assessment of initial and repetitive yields. In addition to filling in an assignment sheet, which included retention times and peak areas, participants were asked to provide specific details about the parameters used for the sequencing run. References for some of the modified amino acid elution characteristics were provided and the participants had the option of viewing a list of the modified amino acids present in the peptide at the ESRG Web site. The ABRF ESRG 2005 sample is the seventeenth in a series of studies designed to aid laboratories in evaluating their abilities to obtain and interpret amino acid sequence data.

A light-initiated, dark oxidation of bacteriochlorophyll from reaction center particles.

When reaction center particles from Rhodopseudomonas spheroides strain R26 are illuminated and then extracted with methanol, about one-third of the extracted bacteriochlorophyll slowly becomes oxidized, The oxidation does not occur under anaerobic conditions or in the absence of the detergent lauryldimethylamine oxide. Alkaline conditions also prevent the reaction. A dark interval between illumination and extraction delays the onset of bacteriochlorophyll oxidation in a predictable way. These results are consistent with the hypothesis that illumination generates a reaction initiator which is fairly stable in methanol but decays with a half-life of about 4.5 min in reaction center particles after illumination ceases.

Effects of oxidants and reductants on the efficiency of excitation transfer in green photosynthetic bacteria.

The efficiency of energy transfer in chlorosome antennas in the green sulfur bacteria Chlorobium vibrioforme and Chlorobium limicola was found to be highly sensitive to the redox potential of the suspension. Energy transfer efficiencies were measured by comparing the absorption spectrum of the bacteriochlorophyll c or d pigments in the chlorosome to the excitation spectrum for fluorescence arising from the chlorosome baseplate and membrane-bound antenna complexes. The efficiency of energy transfer approaches 100% at low redox potentials induced by addition of sodium dithionite or other strong reductants, and is lowered to 10-20% under aerobic conditions or after addition of a variety of membrane-permeable oxidizing agents. The redox effect on energy transfer is observed in whole cells, isolated membranes and purified chlorosomes, indicating that the modulation of energy transfer efficiency arises within the antenna complexes and is not directly mediated by the redox state of the reaction center. It is proposed that chlorosomes contain a component that acts as a highly quenching center in its oxidized state, but is an inefficient quencher when reduced by endogenous or exogenous reductants. This effect may be a control mechanism that prevents cellular damage resulting from reaction of oxygen with reduced low-potential electron acceptors found in the green sulfur bacteria. The redox modulation effect is not observed in the green gliding bacterium Chloroflexus aurantiacus, which contains chlorosomes but does not contain low-potential electron acceptors.

Quality of life in long-term survivors of testicular cancer: a population-based case-control study.

PURPOSE: To evaluate quality of life and social problems in long-term survivors of testicular cancer. PATIENTS AND METHODS: In 1998, 71 testicular cancer survivors (cases) identified from the Calvados General Tumor Registry were enrolled onto a case-control study. One hundred nineteen healthy control subjects (controls), matched by age and location of residence, were selected at random from electoral rolls. Three self-administered questionnaires were used: two health-related quality-of-life questionnaires (Short Form-36 and European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 core questionnaires) and one life situation questionnaire. Specific questions concerning sexuality were also added. RESULTS: With a mean follow-up of 11 years, health-related quality-of-life scores did not differ significantly between cases and controls, nor did general symptom scores. Psychosocial problems were reported equally by cases and controls. Cases reported more modification of sexual life (P =.04) with decreased sexual enjoyment (P <.01), decreased desire (P =.02), and infertility (P <.01). Cases did not report more divorce than controls; they reported fewer changes in relationships with friends (P =.03). Although a similar proportion of cases and controls were at work, cases expressed less ambitious professional plans (P =.002). Cases had greater difficulty in borrowing from banks (P <.001). CONCLUSION: French long-term survivors of testicular cancer do not express more impairment of health-related quality of life or familial or professional life in comparison with healthy men. They did have more sexual life problems and found difficulty in borrowing from banks. This information should be used by practitioners to help their patients cope with their disease and return to normal life.

Radiation therapy of carcinomas of the skin of nose and nasal vestibule: a report of 1676 cases by the Groupe Europeen de Curiethérapie.

A retrospective multicentric analysis of the results of irradiation of 1676 carcinomas of the skin of the nose and nasal vestibule was performed by the Groupe Européen de Curiethérapie (Tunis, May 1986). Overall local control was 93% with a minimum follow-up of 2 years. Local control is dependent on the tumor size (diameter less than 2 cm: 96%, 2-3.9 cm: 88%, greater than or equal to 4 cm: 81%), and tumor site (external surface of the nose: 94%, vestibule: 75%). Local control was independent of histology for smaller tumors, but for those larger than 4 cm, basal cell carcinomas were more frequently controlled than squamous cell carcinomas. Recurrent tumors are less frequently controlled than those being treated for the first time (88 vs. 95%). There were few complications and cosmetic results were generally satisfactory. The results of implantation, orthovoltage and megavoltage irradiation are compared with respect to local control, complication rate and cosmesis. Implantation is usually the treatment of choice for vestibular tumors but for those of the external surface the choice depends on the tumor diameter. Implantation and orthovoltage irradiation are equivalent for tumors less than 2 cm. For those from 2 to 3.9 cm, the results of orthovoltage irradiation may be satisfactory in a selected population but implantation is usually more suitable for tumors with rapidly changing contour.(ABSTRACT TRUNCATED AT 250 WORDS)

Is prophylactic para-aortic irradiation worthwhile in the treatment of advanced cervical carcinoma? Results of a controlled clinical trial of the EORTC radiotherapy group.

From November 1977 to July 1981, 441 patients with cervical carcinoma were randomized between pelvic irradiation and pelvic and para-aortic irradiation. Included were patients with stage I and IIB with proximal vaginal and/or parametrial involvement with positive pelvic lymph nodes either on lymphangiogram or at surgery, and stage IIB with distal vaginal and/or parametrial involvement and III regardless of pelvic node status on lymphangiogram. Patients with clinically or surgically involved para-aortic nodes were not included. The external beam dose to the para-aortic area was fixed at 45 Gy. There was no statistically significant difference between the two treatment arms in terms of local control, overall distant metastases and survival with no evidence of disease (NED), although the incidence of para-aortic metastases and distant metastases without tumor at pelvic sites was significantly higher in patients receiving pelvic irradiation alone (pelvic group). The 4-year NED survival rate was 51%. The incidence of severe digestive complications was significantly higher in patients receiving para-aortic irradiation (para-aortic group). Routine para-aortic irradiation for all high risk patients with cervical carcinoma is of limited value, but patients with a high probability of local control can benefit from extended field irradiation, despite an increase in severe digestive complications.

Prostate cancer treated by radiotherapy: a multivariate study.

According to respective proportions of evolutive status groups, results of multivariate studies are difficult to interpret. Among the 1099 cases of local form of prostate cancer, treated by radiotherapy from 1975 to 1982 in 16 French Anticancer Institutes, we can observe two homogeneous status groups of patients: disease-free survivors (285 cases) and patients who died of prostate cancer (278 cases). These correspond to 51% of the whole population. Among other things, they are comparable in size, for age at the beginning of radiotherapy and for delay between histologic diagnostic and radiotherapy. We chose to analyse them using multivariate analysis. To take survival into account, we used a Cox model and Kaplan-Meier curves; the group deceased of prostate cancer was further analyzed by a tree-structured regression method. The Cox model and the Kaplan-Meier curves confirmed two main explicative factors: Stage (p < 0.0001) and tumor grade (p < 0.001). Poorer evolution occurs in extracapsular forms and grade I has better survival than others. The tree-structured regression method indicates two other pejorative factors: hormonotherapy prior to radiotherapy and the presence of cardiovascular pathology. Though the pelvic dose does not appear to be a main explicative factor, it seems to improve survival and delay between radiotherapy and recurrence or metastasis in some categories of cases. Other factors such as tumor dose, age and delay between diagnosis and radiotherapy were not found to be significant. These results cannot be extended to the whole population for which they do not constitute a predictive study. We consider them as "baseline data".(ABSTRACT TRUNCATED AT 250 WORDS)

A model for recording mercury release from an amalgam surface.

The release rate of mercury from a conventional, a dispersed phase or a spherical high copper content amalgam under static conditions in stimulated or artificial saliva has been measured and found to decrease approximately exponentially with time. A higher initial release rate was observed for mercury in stimulated saliva than in artificial saliva during the in vitro experiments. In a pilot study corrosion current was recorded from an amalgam specimen immersed in saliva, and was found to decrease approximately exponentially with time. After brushing the surface of the amalgam during two subsequent periods, in a manner similar to toothbrushing, an increase in corrosion current was measured indicating the removal of loosely bound corrosion products. A model describing the periodical mercury release from an amalgam surface has been proposed.

Metal release from dental biomaterials.

Levels of corrosion products released from dental alloys in natural or synthetic saliva, i.e. from amalgams, cobalt, gold, nickel, iron, or titanium based alloys have been surveyed. The amounts of Ag, Au, Cd, Co, Cr, Cu, Hg, Mo, Ti or Ni released from such alloys, either in vitro or in vivo during animal tests or during clinical usage have been compiled. The quantities released have been adapted to a 'standard restored man' with a specified number of restorations or a specified construction with a defined surface area, and compared to man's food and drink intake of similar elements. This was done as one approach to a security analysis of wearing dental alloys. In view of the assessment of extensive corrosion testing using electrochemical methods, rather scarce information seems presently available pertinent to release kinetics of specific elements in various biological environments like saliva or saliva substitutes. Several examinations indicate that mercury released from amalgams could be a substantial part of man's daily intake of this element, e.g. in the initial period following insertion, as well as on a long-term basis. From a copper amalgam cadmium could be released in vitro in amounts close to food and drink intake. The mercury release from the amalgam surface seems to be strongly influenced by the interaction of mechanical forces, e.g. chewing, and seems to be released according to a cyclic pattern. From a base metal alloy with high nickel content nickel could be released in vitro at the same level as from food and drink intake. However, from cobalt based alloys the nickel release seems insignificant in this context. The elemental release from the other alloys seemed to be far below the intake of similar elements from food and drink. Release under static and dynamic conditions has been discussed.

Corrosion of a stainless steel with low nickel content under static conditions.

The pattern of selective release of chromium, iron and molybdenum from a non-prepassivated ferritic stainless steel, with the designation SS 2326 and a nickel content of 0.3%, has been measured in artificial saliva under static conditions using nuclear tracer and ESCA techniques. The ICP-method was used to detect nickel which was found only in one case. The release rate of chromium, iron and molybdenum showed a strong time-dependent decrease as a consequence of chromium enrichment in the surface. This was shown by ESCA-measurements. This kinetics is characteristic for a passivating alloy i.e. self passivation. In view of the low nickel content of the SS 2326 steel compared to conventional stainless steel, the material may possess potential for use as a dental or orthopaedic biomaterial.

Liarozole--a novel treatment approach for advanced prostate cancer: results of a large randomized trial versus cyproterone acetate. Liarozole Study Group.

OBJECTIVES: To compare the efficacy of oral liarozole, the first retinoic acid metabolism-blocking agent (RAMBA) to be developed as differentiation therapy for human solid tumors, with that of cyproterone acetate (CPA), an antiandrogen for the treatment of metastatic prostate cancer. Liarozole promotes differentiation of cancer cells by increasing the intratumoral levels of retinoic acid. METHODS: A total of 321 patients with metastatic prostate cancer in relapse after first-line endocrine therapy entered a Phase III international multicenter study (recruitment from February 1992 to August 1994) comparing liarozole (300 mg two times daily) with CPA (100 mg two times daily). RESULTS: Accounting for differences in baseline prognostic factors, the adjusted hazard ratio for survival was 0.74 in favor of liarozole (P = 0.039), indicating a 26% lower risk of death than in patients treated with CPA. Median crude (unadjusted) survival time was the same in the liarozole group as in the CPA group (10.3 months). More patients showed a PSA response (at least 50% reduction in PSA from baseline) when treated with liarozole (20%) than with CPA (4%) (P < 0.001). Prostate-specific antigen (PSA) responders had a median survival benefit of 10 months over nonresponders, irrespective of treatment (hazard ratio 0.43; P = 0.0018). PSA response was apparent within 3 months in approximately 90% of patients who responded. Pain improved more in the liarozole group than in the CPA group (P = 0.03). PSA responders had lower median pain scores than nonresponders (1.7 versus 2.5) and better quality of life (median Functional Living Index-Cancer score 108 versus 98) at end point, ie, treatment discontinuation, as well as throughout the treatment period. Among the most frequently occurring adverse events in the liarozole group were dry skin (51% of patients), pruritus (25%), rash (16%), nail disorders (16%), and hair loss (15%). These adverse events were generally mild to moderate in severity and did not affect the overall quality of life score. There were no detectable effects of either treatment on vital signs such as blood pressure, heart rate, electrocardiogram, and body weight. CONCLUSIONS: Liarozole is superior to CPA in terms of PSA response, PSA progression, and survival, and is capable of maintaining patients' quality of life. The observed adverse events were mild to moderate in nature. These results show that liarozole is a possible treatment option after first-line endocrine therapy has failed.

Redox regulation of energy transfer efficiency in antennas of green photosynthetic bacteria.

The efficiency of energy transfer from the peripheral chlorosome antenna structure to the membrane-bound antenna in green sulfur bacteria depends strongly on the redox potential of the medium. The fluorescence spectra and lifetimes indicate that efficient quenching pathways are induced in the chlorosome at high redox potential. The midpoint redox potential for the induction of this effect in isolated chlorosomes from Chlorobium vibrioforme is -146 mV at pH 7 (vs the normal hydrogen electrode), and the observed midpoint potential (n = 1) decreases by 60 mV per pH unit over the pH range 7-10. Extraction of isolated chlorosomes with hexane has little effect on the redox-induced quenching, indicating that the component(s) responsible for this effect are bound and not readily extractable. We have purified and partially characterized the trimeric water-soluble bacteriochlorophyll a-containing protein from the thermophilic green sulfur bacterium Chlorobium tepidum. This protein is located between the chlorosome and the membrane. Fluorescence spectra of the purified protein indicate that it also contains groups that quench excitations at high redox potential. The results indicate that the energy transfer pathway in green sulfur bacteria is regulated by redox potential. This regulation appears to operate in at least two distinct places in the energy transfer pathway, the oligomeric pigments in the interior of the chlorosome and in the bacteriochlorophyll a protein. The regulatory effect may serve to protect the cell against superoxide-induced damage when oxygen is present. By quenching excitations before they reach the reaction center, reduction and subsequent autooxidation of the low potential electron acceptors found in these organisms is avoided.

Left atrial and right ventricular cardiac myxoma. A case report.

A case is presented with a tumour in the left atrium as well as in the right ventricle. During the initial investigation of the atrial myxoma, the ventricular tumour was overlooked and a second operation was necessary. Once the diagnosis of myxoma is made, a second synchronous tumour should always be carefully sought.

Förster energy transfer in chlorosomes of green photosynthetic bacteria.

Energy transfer properties of whole cells and chlorosome antenna complexes isolated from the green sulfur bacteria Chlorobium limicola (containing bacteriochlorophyll c), Chlorobium vibrioforme (containing bacteriochlorophyll d) and Pelodictyon phaeoclathratiforme (containing bacteriochlorophyll e) were measured. The spectral overlap of the major chlorosome pigment (bacteriochlorophyll c, d or, e) with the bacteriochlorophyll a B795 chlorosome baseplate pigment is greatest for bacteriochlorophyll c and smallest for bacteriochlorophyll e. The absorbance and fluorescence spectra of isolated chlorosomes were measured, fitted to gaussian curves and the overlap factors with B795 calculated. Energy transfer times from the bacteriochlorophyll c, d or e to B795 were measured in whole cells and the results interpreted in terms of the Förster theory of energy transfer.

Determination of uranium in rocks by instrumental activation-analysis using epithermal neutrons.

A rapid non-destructive neutron-activation method for the determination of uranium in rocks is described. The method is based on activation with epithermal neutrons and subsequent measurement of the 74 keV gamma-ray of (239)U. Results given for some standard rocks are in good agreement with literature data. The precision of the method is about 5 % and the limit of detection is of the order of 0.1 ppm in silicate rocks.

Normal concentrations of chromium in serum and urine--a TRACY project.

The validity of "normal" concentrations for total chromium in serum and urine (S-Cr and U-Cr, respectively) in papers published mainly in the last decade were evaluated and graded by two investigators according to TRACY criteria. The results were in close agreement. Because of possible contamination during sampling from stainless steel needles, the description of the sample collection method was considered important. Documentation of analytical quality control was emphasized. The chromium concentrations were categorized according to nonoccupational conditions that could influence the levels. Eighty-seven publications reporting chromium concentrations in blood and 58 on U-Cr were evaluated, 53 dealing with S-Cr and 41 with U-Cr being found suitable for the TRACY project. In selected publications the arithmetic mean values presented for S-Cr and U-Cr in individuals with no known exposure were within the following ranges: 1-3 nmol.1-1 for S-Cr and 2-10 nmol.1-1 or 0.2-1 mumol.mol creatinine-1 for U-Cr.

Antimony in lung, liver and kidney tissue from deceased smelter workers.

Tissue concentrations of antimony in lung, liver, and kidney tissue from a group of deceased smelter workers from northern Sweden have been compared with those of a group of persons without occupational exposure from a nearby area. Neutron activation analysis was used to determine the antimony concentration of lung tissue from exposed workers; these concentrations were 12-fold higher than those of referents (p less than 0.001). For lung tissue there was no tendency towards decreased antimony concentrations with time (up to 20 a) after the cessation of exposure, and this result indicates a long biological half-time. The highest values were found for workers who had worked for many years at the roasters and in the arsenic and selenium departments. There was no significant difference between the antimony concentration of the lung tissue from workers who had died of lung cancer and those of persons who died of other malignancies, cardiovascular disease, or other causes. This finding does not however rule out the possibility of a role for antimony in the etiology of lung cancer among smelter workers since multiple factors may have been operating. The antimony concentration of the liver tissue and the kidney cortex did not differ from the corresponding values of the reference group; this finding indicates either a short biological half-time or insignificance for the systemic distribution of antimony.

Lung retention of antimony and arsenic in hamsters after the intratracheal instillation of industrial dust.

Airborne factory dust (with a volume median diameter of 5.0 micron and a geometric standard deviation of 2.1 micron) from a Swedish copper smeltery contained antimony (Sb) (1.6 weight %) and arsenic (As) (19 weight %). The dust was neutron activated and intratracheally instilled in hamsters. In vivo measurements of lung clearance were undertaken of the radionuclides 76As, 122Sb, and 124Sb. Comparison was made with pure substances of antimony trioxide (Sb2O3) and arsenic trioxide (As2O3). Two phases were recognized in the clearance curves. The approximate half-time for the initial phase was 13 h for As2O3, 20 h for arsenic dust, about 40 h for Sb2O3, and about 30 h for antimony dust. The second phase had an approximate half-time of 20-40 d for Sb2O3 and antimony dust. Because of the short physical half-life of 76As, the second phase of the lung clearance was not possible to follow for AS2O3 and arsenic dust. The observed differences in clearance were primarily related to the solubility of the dust particles in saline, while particle size seemed to be less important in this instillation experiment. The low solubility of antimony in factory dust combined with a long biological half-time may be of importance in explaining the observed lung accumulation of antimony in exposed workers. The greater solubility and shorter biological half-time restricted the lung retention of arsenic.

International project for producing reference values for concentrations of trace elements in human blood and urine--TRACY.

In assessing the concentrations of toxic metals, such as cadmium, chromium, and mercury, in human blood and urine samples to determine whether they are abnormal or not, reliable reference values are needed from populations of nonoccupationally exposed subjects. Numerous publications present concentrations claimed to be typical for the study populations, but they can differ by up to an order of magnitude for a particular element. This is the consequence of general problems that are related to the definition of the reference groups, and the sampling and analytical procedures used, and that make it difficult to define typical and unbiased values. An international group of experts now establishes criteria and procedures to evaluate publications containing information on the concentrations of metals in tissues and body fluids for reference populations. These evaluations have been compiled in a data base (TRACY).