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Evidence for the existence of hypothetical proteins in human bronchial epithelial, fibroblast, amnion, lymphocyte, mesothelial and kidney cell lines.

The human genome maybe limited to about 30000 genes whereas the human proteome may be represented by a rough estimate of one million proteins. A legion of proteins have been described and information about these structures are readily available in data banks. There remains, however, a large series of unknown or hypothetical proteins (HPs). Many of them have been predicted from nucleic acid sequences only and are therefore named predicted or HPs. Carrying out "protein hunting" by generating large maps of human cell lines, we aimed to find and identify HPs and provide an analytical tool thereof. Cell lysates from human bronchial epithelial, fibroblast, amnion, lymphocyte, mesothelial and kidney cell lines were prepared and proteins run on two-dimensional gel-electrophoresis (2DE) with in-gel digestion and mass spectrometrical analysis using the MALDI-TOF principle.16 HPs were found in these cell lines and some show cell-specific expressional patterns. HPs belong to several protein classes including structural, signaling, transcriptional/translational, chaperone-related and others. We furthermore provide analytical data i.e. pIs that were often different from predicted values in data banks.A list of HPs has been shown to really exist in several human cell lines thus contributing to knowledge on protein machineries and cascades. Observed and predicted pI values are given representing an analytical tool along with unambiguous identification of protein spots by mass spectrometry independent of antibody availability and specificity thus complementing established methods.

Amnion↗

Lansoprazole: an update of its place in the management of acid-related disorders.

Lansoprazole is an inhibitor of gastric acid secretion and also exhibits antibacterial activity against Helicobacter pylori in vitro. Current therapy for peptic ulcer disease focuses on the eradication of H. pylori infection with maintenance therapy indicated in those patients who are not cured of H. pylori and those with ulcers resistant to healing. Lansoprazole 30 mg combined with amoxicillin 1g, clarithromycin 250 or 500mg, or metronidazole 400 mg twice daily was associated with eradication rates ranging from 71 to 94%, and ulcer healing rates were generally >80% in well designed studies. In addition, it was as effective as omeprazole- or rabeprazole-based regimens which included these antimicrobial agents. Maintenance therapy with lansoprazole 30 mg/day was significantly more effective than either placebo or ranitidine in preventing ulcer relapse. Importantly, preliminary data suggest that lansoprazole-based eradication therapy is effective in children and the elderly. In the short-term treatment of patients with gastro-oesophageal reflux disease (GORD), lansoprazole 15, 30 or 60 mg/day was significantly more effective than placebo, ranitidine 300 mg/day or cisapride 40 mg/day and similar in efficacy to pantoprazole 40 mg/day in terms of healing of oesophagitis. Lansoprazole 30 mg/day, omeprazole 20 mg/day and pantoprazole 40 mg/day all provided similar symptom relief in these patients. In patients with healed oesophagitis. 12-month maintenance therapy with lansoprazole 15 or 30 mg/day prevented recurrence and was similar to or more effective than omeprazole 10 or 20 mg/day. Available data in patients with NSAID-related disorders or acid-related dyspepsia suggest that lansoprazole is effective in these patients in terms of the prevention of NSAID-related gastrointestinal complications, ulcer healing and symptom relief. Meta-analytic data and postmarketing surveillance in >30,000 patients indicate that lansoprazole is well tolerated both as monotherapy and in combination with antimicrobial agents. After lansoprazole monotherapy commonly reported adverse events included dose-dependent diarrhoea, nausea/vomiting, headache and abdominal pain. After short-term treatment in patients with peptic ulcer, GORD, dyspepsia and gastritis the incidence of adverse events associated with lansoprazole was generally < or = 5%. Similar adverse events were seen in long-term trials, although the incidence was generally higher (< or = 10%). When lansoprazole was administered in combination with amoxicillin, clarithromycin or metronidazole adverse events included diarrhoea, headache and taste disturbance. In conclusion, lansoprazole-based triple therapy is an effective treatment option for the eradication of H. pylori infection in patients with peptic ulcer disease. Preliminary data suggest it may have an important role in the management of this infection in children and the elderly. In the short-term management of GORD, lansoprazole monotherapy offers a more effective alternative to histamine H2-receptor antagonists and initial data indicate that it is an effective short-term treatment option in children and adolescents. In adults lansoprazole maintenance therapy is also an established treatment option for the long-term management of this chronic disease. Lansoprazole has a role in the treatment and prevention of NSAID-related ulcers and the treatment of acid-related dyspepsia; however, further studies are needed to confirm its place in these indications. Lansoprazole has emerged as a useful and well tolerated treatment option in the management of acid-related disorders.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Advantages and drawbacks of immunoaffinity columns in analysis of mycotoxins in food.

A number of countries are setting legislations on mycotoxins. In order to reduce dispute between importing and exporting countries, the analytical data should be as comparable as possible, especially when levels are close to the regulatory limits. The present trend in the analysis of mycotoxins is to use immunoaffinity column (IAC) as a clean-up and enrichment technique, and Association of Official Analytical Chemists and European Union have validated methods which address a few food commodities. This study describes our experience using both conventional and IAC approaches in the analysis of three mycotoxins. Aflatoxins (AFs): Aflatoxin G1 has been detected by liquid-liquid partitioning methods with HPLC detection as false-positive in some maize. On IACs, this compound behaves as an AF, lowering the amount of the AFs trapped. The problem was solved using either TLC or HPLC with detection in the Kobra cell. Depending on the additives to food during the processing and cooking, the AFs might appear as an opened ring not recognised by the antibody. Fumonisins (FB): Compounds interfering with the FB's antibodies were also observed while analysing breakfast cereals leading to underestimation of FB. Ochratoxin A (OTA): Depending on the food composition and extraction techniques, OTA is underestimated with IAC in some breakfast cereals and coffee. These data strengthen the necessity to validate methods using IAC for each complex matrix.

Aflatoxin B1↗

[Vertically transmitted bacteremias: to treat or not to treat?].

OBJECTIVE: To analyze the incidence, etiology and management of infants born in 1977 with vertically transmitted bacteremia or suspected early neonatal sepsis. PATIENTS AND METHODS: The total number of newborn infants in this period was 2,365. We revised the clinical histories of the infants diagnosed with bacteremia and classified them into two groups: a) those with vertically transmitted bacteremias, according to the recommendations of the Castrillo Group, and b) those with suspected early onset neonatal sepsis in whom blood culture was positive and analytical data suggested bacterial infection but who showed no clinical symptoms of vertically transmitted sepsis. Birthweight, sex, gestational age, risk factors for neonatal infection, clinical signs and laboratory tests suggestive of bacterial infection and microbiological agents were analyzed. The clinical and analytical evolution of the treated and untreated newborn infants was studied. RESULTS: The newborn infants were diagnosed with vertically transmitted bacteremia (an incidence of 4.2x1,000 live newborn infants) and 17 were diagnosed with suspected early onset neonatal sepsis (7.8x1,000 live newborns). All the infants had risk factors for neonatal sepsis. The most common of them was prolonged membrane rupture (>- 18 hours) due to which sepsis screening was carried out (hemogram, C-reactive protein at 12 and 36-48 hours of life, and blood culture). In both groups the most commonly isolated microorganism was group B streptococcus, which was found in 30% of vertically transmitted bacteremias and in 41.2% of suspected early onset neonatal sepsis. All the newborn infants with suspected sepsis and two with vertically transmitted bacteremia were treated without incident. The remaining eight infants with untreated vertically transmitted bacteremia were followed-up clinically ana analytically for one year, and remained asymptomatic. CONCLUSIONS: The most common microorganism in vertically transmitted bacteremia and suspected early onset neonatal sepsis was group B streptococcus. None of the untreated infants developed late sepsis or meningitis. Our findings suggest that non-treatment of asymptomatic infants with vertically transmitted bacteremias is appropriate as long and close clinical surveillance is maintained.

Age Factors↗

Multicountry estimation of dietary boron intake.

Dietary Reference Intakes are not yet established for boron (B), a naturally occurring trace element in the human diet. Estimated dietary B intakes provide useful information for planning and assessing diets in healthy populations. The countries selected for this study represent a wide variety of dietary patterns and have adequate nutrient databases (with the exception of B) and food consumption data. Large-scale nationwide survey data were provided by the US (1989-1991) and Germany (1985-1989). Survey data from rural agricultural communities of Mexico and Kenya were provided by the Human Nutrition Collaborative Research Support Program (1983-1986). A B nutrient database was created to include B concentrations for the foods consumed in each country. It incorporates B analytical data from various sources in the US, Finland, UK, Italy, Japan, and China. Each person's average daily B intake was estimated by linking the B database with the survey food records. Average dietary intake estimates were then generated for various age and sex groups. The estimates for adults in the US, Germany, Mexico and Kenya, respectively, are 1.11, 1.72, 2.12, and 1.95 mg B/d for males and 0.89, 1.62, 1.75, and 1.80 mg B/d for females. Foods that are major contributors to the B intake of each country were also identified.

Adolescent↗

Use of a sample translation technique to minimize adverse effects of laser irradiation in surface-enhanced Raman spectrometry.

Surface-enhanced Raman spectroscopy (SERS) has proven to be a very powerful tool in the analysis of a wide range of compounds. However, continuous irradiation of the laser beam over the SERS substrate can promote the gross decomposition of the sample analytes and significantly broaden and diminish the intensities of observed spectral bands. In addition, the incident radiation can promote thermal or photolytic fragmentation of analytes, thereby altering the observable bands and possibly leading to a misinterpretation of analytical data. Finally, chemical or morphological changes in the SERS substrate are possible. This work presents the use of a sample translation technique (STT) as a means to minimize these adverse effects. By spinning the sample rapidly, the effective residence time of analytes and substrate within the irradiated zone is dramatically decreased without reduction of spectral acquisition time or the density of analyte in the zone. The technique is studied by acquiring SERS spectra of Naproxen USP, riboflavin, folic acid, Rhodamine 6G, and 4-aminothiophenol using silver islands on glass and silver-poly(dimethylsiloxane) composite substrates under various spinning and stationary conditions. In all cases, spectra show improvements upon spinning at laser powers as low as 4.2 (+/- 0.1) mW. Specific differences in the appearance of the spectra and the potential use of STT for improved SERS qualitative and quantitative determinations are presented.

Aniline Compounds↗

Automated storage of gas chromatography-mass spectrometry data in a relational database to facilitate compound screening and identification.

This paper describes a database containing massspectra from gas chromatography-mass spectrometry(GC-MS) measurements as a tool for easy screening for multiple compounds. In this way additional compounds can be reported from the same run together with routine pesticide monitoring with little effort. The relevant analytical data from the GC-MS measurements are transferred automatically to a database. Search algorithms in the database, containing the US EPA and Dutch NEN GC-MS identification criteria as standard settings, are used to identify compounds in the data. Screening of samples analysed in our laboratory show the ubiquitous presence of--up until now in monitoring largely overlooked--compounds in surface waters in The Netherlands. Most frequently found compounds include TAED (complexing agent), 2-methyl quinoline (industrial solvent), atrazin and desethylatrazin (pesticide and degradation product), caffeine (human consumption), surfinol-104 (anti foaming agent), HHCB (Galaxolide) and AHTN (Tonalide; fragrances). The database can also be used to quickly search a large number of datafiles for rare contaminants. This way, some interesting compounds such as pentoxifilin (a pharmaceutical) and Irgarol 1051 (an antifouling compound) were found.

Database Management Systems↗

Dietary fat and cancer: consistency of the epidemiologic data, and disease prevention that may follow from a practical reduction in fat consumption.

International variations and national time trends in disease rates suggest major associations between dietary fat and several important cancers. In contrast, case-control and cohort studies of dietary fat in relation to the same cancers generally report weak associations, or have failed to detect any association with fat intake. This study was undertaken in an attempt to understand the apparent discrepancy between these observations. The results provide an insight into the magnitude of cancer risk reduction that may follow from a practical reduction in dietary fat. Regression analyses of international variations in cancer incidence rates were used to estimate relative risks (RR) as a function of fat intakes for both males and females. These analyses focused on cancers of the breast, colon, rectum, ovary, and endometrium in females, and colon, rectum, and prostate cancers in males. Ages 55-69 and 30-44 were considered in order to compare RR estimates between an older and younger age group, and between post- and pre-menopausal women. Corresponding RR estimates were also calculated, based on the regression of changes in disease rates from the mid-1960s to 1980 on changes in dietary fat, using data from several countries. A strong degree of consistency with the RR estimates from international comparisons was observed. The international regression analyses were also used to project changes in cancer rates among Japanese migrants to the United States. A high level of consistency with the observed disease-rate changes was noted. Similarly, the international data analyses were used to project RRs for the fat intake categories used in specific case-control and cohort studies, while acknowledging measurement error in individual dietary assessment. Although certain exceptions are noted, considerable consistency was found between the aggregate and analytic data results, leaving open the strong possibility that a practical reduction in dietary fat could result in a major reduction in the incidence of several prominent cancers in the United States and in other nations having high fat consumption.

Adult↗

Inactivation of Bacillus subtilis glutamine synthetase by metal-catalyzed oxidation.

Instability of Bacillus subtilis glutamine synthetase in crude extracts was attributed to site-specific oxidation by a mixed-function oxidation, and not to limited proteolysis by intracellular serine proteases (ISP). The crude extract from B. subtilis KN2, which is deficient in three intracellular proteases, inactivated glutamine synthetase similarly to the wild-type strain extract. To understand the structural basis of the functional change, oxidative modification of B. subtilis glutamine synthetase was studied utilizing a model system consisting of ascorbate, oxygen, and iron salts. The inactivation reaction appeared to be first order with respect to the concentration of unmodified enzyme. The loss of catalytic activity was proportional to the weakening of subunit interactions. B. subtilis glutamine synthetase was protected from oxidative modification by either 5 mM Mn2+ or 5 mM Mn2+ plus 5 mM ATP, but not by Mg2+. The CD-spectra and electron microscopic data showed that oxidative modification induced relatively subtle changes in the dodecameric enzyme molecules, but did not denature the protein. These limited changes are consistent with a site-specific free radical mechanism occurring at the metal binding site of the enzyme. Analytical data of the inactivated enzyme showed that loss of catalytic activity occurred faster than the appearance of carbonyl groups in amino acid side chains of the protein. In B. subtilis glutamine synthetase, the catalytic activity was highly sensitive to minute deviations of conformation in the dodecameric molecules and these subtle changes in the molecules could be regarded as markers for susceptibility to proteolysis.

Bacillus subtilis↗

Advances in tumor subclone formation and mechanisms of growth and invasion.

Tumor subclones refer to distinct cell populations within the same tumor that possess different genetic characteristics. They play a crucial role in understanding tumor heterogeneity, evolution, and therapeutic resistance. The formation of tumor subclones is driven by several key mechanisms, including the inherent genetic instability of tumor cells, which facilitates the accumulation of novel mutations; selective pressures from the tumor microenvironment and therapeutic interventions, which promote the expansion of certain subclones; and epigenetic modifications, such as DNA methylation and histone modifications, which alter gene expression patterns. Major methodologies for studying tumor subclones include single-cell sequencing, liquid biopsy, and spatial transcriptomics, which provide insights into clonal architecture and dynamic evolution. Beyond their direct involvement in tumor growth and invasion, subclones significantly contribute to tumor heterogeneity, immune evasion, and treatment resistance. Thus, an in-depth investigation of tumor subclones not only aids in guiding personalized precision therapy, overcoming drug resistance, and identifying novel therapeutic targets, but also enhances our ability to predict recurrence and metastasis risks while elucidating the mechanisms underlying tumor heterogeneity. The integration of artificial intelligence, big data analytics, and multi-omics technologies is expected to further advance research in tumor subclones, paving the way for novel strategies in cancer diagnosis and treatment. This review aims to provide a comprehensive overview of tumor subclone formation mechanisms, evolutionary models, analytical methods, and clinical implications, offering insights into precision oncology and future translational research.

Humans↗

The amelogenin problem: a comparison of purified enamel matrix proteins.

Using a combination of gel filtration and DEAE-cellulose chromatography, together with small-scale preparative polyacrylamide gel electrophoresis, we isolated five proteins (amelogenins) from demineralized bovine fetal dental enamel matrix. These purified proteins were characterized by amino acid analysis and gel electrophoresis. Comparisons of these data with those of other workers suggest that, although there are similarities in the published data between components of comparable electrophoretic mobility, there are gross differences in the reported amino acid compositions. It is suggested that these differences are due not to separative problems arising from reversible aggregations, but to inadequate comparisons of the electrophoretic and amino acid analytical data.

Amelogenesis↗

Development of a quality-assessed agrichemical database for monitoring anthropogenic impacts on ground-water quality.

The Quality-Assessed Agrichemical Contaminant Database for Nebraska Ground Water is a unique repository of nitrate and pesticide data collected by federal, state, and local agencies. Each contaminant concentration in the database has been evaluated based upon well-defined criteria that address completeness of the well-attribute data, analytical method and field and laboratory quality control practices and assigned to one of five quality levels. The quality assessment level always accompanies the contaminant concentration so that the end-user knows the quality assurance effort expended in the acquisition of the data, can select comparable data, and choose data whose quality assurance effort is commensurate with project objectives. The database can be viewed and queried on-line; downloaded in its entirety; or imported to a spreadsheet or a geographic information system. Setting criteria for data quality and assessing the level of quality have resulted in significant increases in the percentages of high quality (Levels 3-5) nitrate and pesticide data. These high quality data presently constitute 52% of the nitrate and 55% of the pesticide data.

Agrochemicals↗

[Severe chickenpox pneumonia].

OBJECTIVE: Retrospective study of the epidemiological, clinical, diagnostic and therapeutic data corresponding to adult patients with chickenpox pneumonia that needed admission in the Intensive Care Unit (ICU) in the last 10 years. MATERIAL AND METHODS: The diagnosis was established through clinicoradiological criteria along the chickenpox progress. Assessed parameters are underlying diseases, pregnancy, smoking habits, analytical data, progress toward adult respiratory distress syndrome (ARDS), need for mechanical ventilation, and pharmacological treatment. RESULTS: They were studied 8 patients (7 women and 1 man) with an average age of 30 years (range: 25-38). Only one of the patients showed underlying pathology (chronic B hepatitis and alcoholism), and another patient was pregnant with 24 weeks of gestation. All of them were smokers. The respiratory symptoms appeared between the second and fourth day from the onset of the exanthema. All the patients showed dyspnea and in seven of them it was associated to dry cough. The chest x-ray at the time of the admission showed in all the patients a bilateral interstitial infiltrate of basal predominance. In 7 of the 8 cases less than 150,000 platelets/mm3 were observed, in 5 patients an increase of transaminases was detected, and all the patients showed elevation of LDH. In 6 patients hypoxemia (PaO2<60 mmHg) was detected with need in 3 of them of orotracheal intubation and mechanical ventilation. These 3 patients progressed toward ARDS. All the cases studied were treated with intravenous aciclovir. None of the hospitalized patients died. CONCLUSIONS: Chickenpox pneumonia is a serious complication that can force hospitalization in the ICU. The severity of the disease can range from the observation of asymptomatic radiological alterations up to ARDS. Mortality is high, but is probably declining in recent years due to the early antiviral treatment and to the application of appropriate support measures.

Adult↗

Biomechanical modeling of anterior spine instrumentation in AIS.

This study is part of a larger project regarding the development of a Spine Surgery Simulator (S3), which has shown good results for posterior instrumentation surgeries. The aim was to develop a biomechanical model for the anterior instrumentation of the scoliotic spine. A biomechanical model using flexible mechanism was developed and surgical manoeuvres (instrumentation, rod installation and compression) were reproduced. Validation of the model was done by comparing the results for the instrumented part of the spine to the post-operative data (analytical Cobb angles in the frontal and sagittal planes, plane of maximum deformity, etc.). To date, surgeries of four patients operated by thoracotomy were reproduced. Preliminary results show that anterior instrumentation of the scoliotic spine can be adequately modelled using pre-operative geometric data and using mechanical properties from literature. Once validated with a larger sample of cases, the anterior instrumentation model could be implemented into S3 and used by orthopaedic surgeons to test various instrumentation strategies in virtual reality before performing the actual surgery.

Adolescent↗

Determination of pesticides in water by cone-shaped membrane protected liquid phase microextraction prior to micro-liquid chromatography.

A new sample pre-treatment technique termed cone-shaped membrane liquid phase microextraction (CSM-LPME) was developed and combined with micro-liquid chromatography (micro-LC) for the determination of selected pesticides in water samples. Four pesticides (hexaconazole, procymidone, quinalphos and vinclozolin) were considered as target analytes. Several important extraction parameters such as types of extraction solvent, agitation rate, pH value, total exposure time and effect of salt and humic acids were optimized. Enrichment factors of > 50 folds were easily achieved within 20 min of extraction. The analytical data demonstrated relative standard deviations for the reproducibility of the optimized CSM-LPME method ranging from 6.3 to 7.5%. The correlation coefficients of the calibration curves were at least 0.9995 across a concentration range of 2-100 microg/L. The detection limits for all the analytes were found to be in the range of 1.1-1.9 microg/L.

Analytic Sample Preparation Methods↗

The role of science in Codex standards.

The key principle in the development of Codex standards is to ensure that the decision-making process is based on sound science. The approval for the use of food additives and other chemicals in foods should be made only when adequate scientific data is available. The same principle applies to the development of maximum tolerable (or residue) levels for contaminants in foods. Using the General Standards for Contaminants and Toxins in Foods as an example, the following criteria are considered, when developing recommendations and standards: Toxicological information; Analytical data; Intake data; Fair trade considerations; Technological considerations; and Risk assessment and risk management considerations. The Codex Alimentarius Commission has repeatedly emphasized the use of risk analysis approach, in particular the use of risk assessment in conducting the safety evaluation of food additives agricultural and veterinary chemicals and environmental and industrial contaminants in foods. In this respect, the well known efforts of the FAO/WHO Joint Expert Committee on Food Additives (JECFA) and the FAD/WHO Joint Meeting on Pesticide Residues (JMPR) provide the much needed information and recommendations for the risk assessment of specific chemicals. The risk assessment approach is also used in the safety evaluation of foods derived from modern biotechnology as well as in the quantitative assessment of microbiological risks in foods.

Agriculture↗

Polycyclic aromatic hydrocarbons in Norwegian forest soils: impact of long range atmospheric transport.

Levels of nine selected polycyclic aromatic hydrocarbons, PAHs, in surface soils from areas in southern and central Norway are presented. Levels in central Norway are generally low, while the southern Norway soils are about ten-fold higher with respect to 4 and 5 ring PAHs. Comparison with air quality data indicates long-range atmospheric transport to be the major source of the excess 4 and 5 ring PAHs in the south. Analyses of peat cores from ombrotrophic bogs support this assumption, and these provide a potentially useful approach for temporal studies of atmospheric PAH deposition. Analytical data for naphthalene in soils depend very much on the sampling and storage of the samples before analysis.

Journal Article↗

Primary epiploic appendagitis: US and CT findings.

A retrospective review is presented of seven cases of epiploic appendagitis, with surgical confirmation in one case. The main clinico-analytical data and the US and CT findings are described, as well as the histopathologic features in the sole case that underwent surgical resection. We also calculated the frequency of this entity in patients undergoing emergency abdominal US on clinical suspicion of diverticulitis. In all seven cases the clinico-analytical evidence was nonspecific (localized acute abdominal pain and slight leukocytosis), mimicking in six cases the clinical presentation of sigmoid diverticulitis and in one case that of acute appendicitis. US imaging findings were characteristic: a hyperechoic mass localized under the point of maximum pain, adjacent to the anterior peritoneal wall and fixed during deep breathing. In none of the cases did color Doppler US show flow. CT findings were also typical and showed a mass with a peripheral hyperattenuated rim surrounding an area of fatty attenuation. Overall 7.1 % of patients investigated to exclude sigmoid diverticulitis finally showed findings of primary epiploic appendagitis. Primary epiploic appendagitis thus shows characteristic US and CT findings that allow its diagnosis and follow-up. This entity is much more frequent than previously reported, especially in patients referred for US to exclude sigmoid diverticulitis.

Abdominal Pain↗