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Results for “Protein profiling”

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PubMed1989

Two-dimensional protein profiles of cultured stromal and epithelial cells from hyperplastic human prostate.

Studies were undertaken to compare and contrast the two-dimensional protein profiles of epithelial and stromal cells from hyperplastic human prostate to establish the protein composition of the two major cellular components of the prostate. Epithelial and stromal cells were isolated from human prostate obtained from patients undergoing open prostatectomy for benign prostatic hyperplasia (BPH). Proteins, isolated from the two cell populations and separated by two-dimensional (2D) electrophoresis, were analyzed by silver staining, fluorography of [35S]-methionine-labeled proteins, and immunoprotein blotting. Isolated prostatic epithelial cells, but not stromal cells, contained cytokeratin polypeptides 5, 6, 7, 8, 13, 14, 15, 16, 17, 18, and 19. Although vimentin could not be identified in silver stained 2D gels and fluorographs of cultured prostatic epithelial cells, a low level of immunoreactivity was noted following immunoblot analysis of epithelial cells proteins by the use of an anti-vimentin polyclonal. Vimentin was prominently expressed in cultured prostatic stromal cells and could be identified on silver stained 2D gels, fluorographs, and immunoblots of stroma-derived proteins. In addition, stromal marker proteins SM1, SM2, and SM3 were identified in 2D gels of stromal cells to distinguish them from epithelial cells. These studies demonstrate (1) the two-dimensional protein profile and cytokeratin polypeptide composition of cultured epithelial cells from hyperplastic human prostate and (2) the 2D protein profile of cultured prostatic stromal cells and identification of specific stromal marker proteins.

Cells, Cultured↗

PubMed1992

Lytic activities, protein profiles and morphologic characteristics of new bacteriophages isolated from canine and human Staphylococcus aureus strains.

The lytic activity, protein profile and morphology of five newly isolated phages from canine Staphylococcus aureus strains and one from a human S. aureus strain were compared with those of selected phages in the international phage sets (IPS). Five canine phages lysed 57 (76.0%) of 75 canine isolates of Staphylococcus aureus from Nigeria at routine test dilution (RTD) while 34 (IPS) phages typed only 31 (41.3%) strains at RTD or/and 100-RTD. The new human phage lysed 11 (14.7%) of 75 strains isolated from human diarrhoea. The new phages were readily propagated, specific in activity and stable during storage at 4 degrees C. Prominent proteins detected by SDS-PAGE indicated similarities between some of the phages but one canine phage was distinctly different, as was its morphology which was an isometric head with a short tail compared to oval heads and long tails which characterized others. IPS phages in the same serologic group had similar protein profiles but no correlation was observed with lytic groups. The use of protein profile and electron micrographs allowed classification of the phages into serogroups. It is concluded that the newly isolated canine phages could be very useful in typing Nigerian canine strains of S. aureus.

Animals↗

PubMed1984

[Changes in the serum protein profile during radiotherapy of the upper respiratory and digestive tracts].

Patients with a cancer of the upper airways or upper gastro-intestinal tract present a state of malnutrition as a result of the disease itself and, more importantly, as a result of its localisation. Loco-regional radiotherapy often leads to an aggravation of this state. The protein profile, consisting of nine serum proteins, was determined each week in 54 patients with cancer of the upper respirato-gastro-intestinal tract receiving radiotherapy. During the course of radiotherapy, the already altered nutritional state of these patients deteriorated further, as shown by a regular and significant downturn in the weight curve. The weekly monitoring of the protein profile showed a gradual and significant decrease in the levels of nutritional proteins (prealbumin, retinol binding protein, transferrin) and immunoglobulins (IgM, IgA) and a small variation in the levels of inflammatory proteins (haptoglobin, orosomucoid, C3 complement fraction, alpha 1-antitrypsin). The protein profile, established on the basis of carefully selected proteins, can provide useful information in the monitoring of a patient's nutritional state.

Adult↗

PubMed1998

Heterogeneity of protein profiles of Helicobacter pylori isolated from individual patients.

BACKGROUND: In order to characterize the diversity of Helicobacter pylori (H. pylori) in infected individuals, 10 colonies of H. pylori were isolated from the gastric juice of 25 patients with gastroduodenal diseases (total 250 isolates). METHODS: Protein profiles of isolates were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Results were confirmed by Western blotting (immunoblotting) test using rabbit antisera against three different strains of H. pylori. RESULTS: The protein profiles of 18 of 25 cases (72%) showed a single type of H. pylori with the same polypeptide pattern. In contrast, heterogeneity in the protein profiles was seen in isolates from seven cases (28%). Two differing H. pylori types with two very different polypeptide patterns were found in 10 isolates from one case. In six patients, the protein profiles of isolates were found to have variations in their polypeptides between molecular weights of 30,000 (30K) and 14K, which are thought to be associated with bacterial membrane protein. In some isolates, a polypeptide band of the 16K was missing. Each of three different antisera confirmed differences among the distinct isolates from individual patients. CONCLUSIONS: These findings suggest that more than one antigenically different strain of H. pylori may exist in same infected individuals.

Animals↗

PubMed2004

Cell-specific protein profiling in Arabidopsis thaliana trichomes: identification of trichome-located proteins involved in sulfur metabolism and detoxification.

Metabolite, protein, and transcript analysis at the cellular level gives unparalleled insight into the complex roles tissues play in the plant system. However, while capillary electrophoresis and PCR amplification strategies make the profiling of metabolites and transcripts in specific cell types possible, the profiling of proteins in small samples represents a bottleneck. Here for the first time protein profiling has been achieved in a specific plant cell type: The application of specific cell sampling and shotgun peptide sequencing (nano LC/MS/MS) resulted in the identification of 63 unique proteins from pooled Arabidopsis trichome cells. A complete S-adenosylmethionine pathway cluster, two S-adenosylmethionine synthase isoforms, a glutathione S-conjugate translocator and other proteins involved in sulfur metabolism and detoxification are shown to be present in these cells, in agreement with previous work done at the level of trichome transcript analysis. The technology described here brings the simultaneous identification and localization of physiologically relevant cellular proteins within reach.

Arabidopsis↗

PubMed2002

Lung-stage protein profile and antigenic relationship between Ascaris lumbricoides and Ascaris suum.

The protein profile and antigenic properties of lung-stage larvae of Ascaris lumbricoides and A. suum were studied using 2-dimensional electrophoresis and immunoblot analysis, respectively. The protein profiles of the 2 parasites were identical except for the presence of only 1 major protein spot specific for each. There was a complete cross-reactivity between the 2 parasites at the immunological level, and no specific antigen was recognized using specific antibody raised against the 2 parasites in rabbits.

Animals↗

PubMed2005

Comparative protein profiles of Salmonella and Escherichia coli.

Protein profiles of selected Salmonella serovars were compared with E. coli to identify genus specific protein(s) for Salmonella. The PDP formed of different Salmonella serovars were compared with E. coli O78 when subjected to SDS-PAGE yielded 11, 15, 15, 11 and 14 bands in S. Bareilly, S. Gallinarum, S. Typhimurium and S. Weltevreden and E. coli O78 respectively. The bands produced were compared with each other. It was found that S. Weltevreden shared 7 bands with E. coli O78, A protein of molecular weight 20.89 kDa was found in all Salmonella serovars, but not in E. coli O78 suggesting its genus specific attribute.

Bacterial Proteins↗

PubMed2000

Hypobiosis induction alters the protein profile of Ostertagia ostertagi (Nematoda: Trichostrongylidae).

The appearance of variations in the protein profile of Ostertagia ostertagi (Stiles, 1892) infective larvae (L3), which were induced by hypobiosis triggering factors, was evaluated by means of SDS-PAGE and densitometric analysis. Area integration analyses of their protein profiles was carried out between 66 and 77 kDa. Important quantitative variations were identified in the protein levels of the induced larvae, where a 5.25 fold increase compared to the control was observed. Two 75.4 and 70 kDa protein bands were found which exceeded the control profile by 4.5 and 44 fold, respectively. This fact suggests that the changes brought about in the process of hypobiosis induction are restricted. This work demonstrates changes at a molecular level corresponding with biological changes induced by conditions causing O. ostertagi hypobiosis.

Animals↗

PubMed1984

[The concept of protein profile].

From their own experience of the simultaneous immunonephelometry of eight serum proteins, the authors propose a definition of protein profile from the point of view both of laboratory technique and interpretation. The assay should be performed quickly and the results expressed diagrammatically in normalised values, in such a way that the relative variation in protein levels can be easily visualised. A protein profile should thus comprise a minimum of proteins to be measured, chosen according to protein physiopathology and the type of abnormality under investigation. Interpretation is based on inter-protein correlations that may appear or disappear depending on the underlying physiopathology. This makes it possible to study inter-protein variation and thus avoid the probabilistic "interpretation" of single proteins taken in isolation. From this approach syndromes can be divided into two groups, elementary or complex. The authors provide examples of each, and propose an interpretation model based on this dichotomy.

Blood Protein Electrophoresis↗

PubMed1992

Effects of maternal marginal zinc deficiency on myelin protein profiles in the suckling rat and infant rhesus monkey.

In the current study, the effects of marginal Zn deficiency on myelin protein profiles in neonatal rats and rhesus monkeys were investigated. Following mating, rats were fed a Zn-adequate diet, ad libitum (50 micrograms Zn/g; 50 Zn AL), or a marginal Zn diet (10 micrograms Zn/g) from day 0 (10 Zn d0) or day 14 (10 Zn d14) of gestation to day 20 postnatal. An additional group of dams was restricted-fed the control diet to the food intake of the 10 Zn d0 group (50 Zn RF). Day 20 pup plasma and liver Zn concentrations in the 10 Zn groups were lower than in the 50 Zn groups. In a parallel experiment, rhesus monkeys were fed a Zn-adequate ad libitum diet (100 micrograms Zn/g) or a marginal Zn diet (4 micrograms Zn/g diet; MZD) throughout gestation and lactation. Day 30 monkey infant plasma and liver Zn levels were similar in the MZD and control groups. Rat brain and monkey brain cortex weights were similar among the dietary groups. The amount of myelin recovered (mg protein/g brain) from day 20 rat pups from the 10 Zn groups was lower than that recovered from the 50 Zn rat pups. Myelin recovery from the MZD and control monkey infants was similar. When myelin protein profiles were characterized, it was found that the percentages of high-molecular-weight (HMW) proteins and Wolfgram protein were higher, whereas the percentages of small and large basic proteins were lower in myelin from the 10 Zn d0 and 50 Zn RF pups compared to the distribution in the 50 Zn AL rat pups. Results for the 10 Zn d0 and 10 Zn d14 pups were similar for all of the parameters studied. The percentage of HMW proteins was higher and that of basic protein lower in myelin from MZD monkey infants compared to the percentage of these proteins in myelin from controls. Although the interpretation of the rat data is complicated because of the anorexia associated with Zn deficiency, the observed changes in monkey myelin protein profiles provide strong evidence that maternal Zn deficiency affects myelination in the offspring.

Animals↗

PubMed1984

The effects of feed form, protein profile, energy level, and gender on broiler performance in warm (26.7 C) environments.

Two dietary energy levels, 3032 and 3109 kcal ME/kg, and four protein levels of approximately 17, 18, 19 and 20%, were used to formulate eight grower-finisher rations used in four feeding regimens used in both mass and crumble form fed to broilers in a warm (26.7 C) environment. The protein levels were established by specific lysine: energy ratios with methionine plus cystine at a constant percentage of lysine. Best overall broiler performance was obtained with a feeding regimen using the higher energy level with high-protein profile in crumble form; feed conversion for this regimen was improved 5.4% and body weight 3.2% over the poorest performing diet, which was the lower energy diet with low-protein profile in mash form. Overall, crumbling improved feed conversion 1.5% and increased body weight 2.2%, the high-protein profile improved feed conversion 2.0%, and increasing metabolizable energy from 3032 to 3109 kcal/kg improved feed conversion 2.2%. Feed conversion for males was 3.3% better than for females.

Animal Feed↗

PubMed1989

Epidemiologic marker system for Citrobacter diversus using outer membrane protein profiles.

Investigations of nursery outbreaks of Citrobacter diversus sepsis and meningitis have been hampered by lack of adequate epidemiologic markers for the organism. We studied outer membrane protein profiles from clinical isolates of C. diversus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine whether this method might be useful in the epidemiologic differentiation of strains. Paired cerebrospinal fluid isolates from each of three separate nursery outbreaks of C. diversus meningitis, paired isolates from the vagina of a postpartum woman and the cerebrospinal fluid of her newborn infant, one isolate from an infant with pneumonia and two from colonized nursery cohorts, and 30 epidemiologically unrelated clinical isolates were included. Eleven distinct profiles were differentiated by the presence or absence of five outer membrane proteins. Complete concordance of profiles was observed for epidemiologically related isolates. Unrelated epidemic strains had outer membrane protein profiles distinct from one another. Biotyping complemented determination of outer membrane protein profiles; the two markers differentiated each of the five epidemic strains from all but one of 30 unrelated nonepidemic isolates. Determination of outer membrane protein profiles is potentially useful in epidemiologic investigations of disease caused by C. diversus.

Bacterial Outer Membrane Proteins↗

PubMed2004

Relationship between interaction sites in the gut, hydrophobicity, mucosal immunomodulating capacities and cell wall protein profiles in indigenous and exogenous bacteria.

AIMS: To investigate whether there is a relationship between interaction sites in the gut, hydrophobicity, mucosal immunomodulating capacities and cell wall protein profiles in lactobacilli, bifidobacteria and enterococci. METHODS AND RESULTS: Hydrophobicity, cell wall protein profiles and sites of interaction in the gut (by using fluorescein isothiocyanate-labelled bacteria) were determined for Lactobacillus casei, L. acidophilus, L. fermentum, Bifidobacterium bifidum, B. animalis and Enterococcus faecalis. We also determined the number of immunoglobulin (Ig)A+, tumour necrosis factor (TNF)alpha+, interleukin (IL)-6+ and IL-10+ cells after oral administration of the above bacteria to BALB/c mice. All strains assessed were found to interact with the sites of induction of the immune response in the gut. No correlation with hydrophobicity was observed. When some strains at certain doses were administered to mice, bacterial translocation to liver was observed. The oral administration of indigenous (104 cells day(-1)) and exogenous (107 cells day(-1)) bifidobacteria and lactobacilli for 5 consecutive days activated the systemic and intestinal mucosal immune response in a strain-specific way, independently whether the strain was indigenous or exogenous in relation to the host. The differences in the immunopotentiating capacity of the various strains might be related to the differences in their cell wall protein profiles. CONCLUSIONS: Indigenous bacteria activated the mucosal immune response at a dose significantly smaller than the one required for probiotic exogenous bacteria. However, probiotic exogenous bacteria can be used at high concentrations in fermented dairy products with a great impact on the immune system, favouring its immunomodulation. SIGNIFICANCE AND IMPACT OF THE STUDY: The immunomodulation capacity of probiotic bacteria is strain specific and independent of the specificity of the host. The ability of certain strains to down-regulate the production and release of IL-6 by IL-10 may have potential implications in their use in cases in which cytokine deregulation or excessive production at the mucosal level can be the cause of tissue damage.

Animals↗

PubMed2000

Efficacy of a potentized homeopathic drug (Arsenicum-Aalbum-30) in reducing cytotoxic effects produced by arsenic trioxide in mice: IV. Pathological changes, protein profiles, and content of DNA and RNA.

OBJECTIVE: To examine if the potentized homeopathic drug Arsenicum Album-30 can help restore the damage produced in protein profiles, DNA and RNA contents in liver and testis as a result of arsenic treatment in mice. DESIGN: Sets of mice were injected with arsenic trioxide, one set was fed with Ars. Alb-30, another with Alcohol-30 and the final set was fed neither. The gel electrophoretic protein profiles and DNA and RNA contents in these three sets were studied. METHODS: Protein profiles were studied by SDS-PAGE method; the DNA and RNA contents were assayed by the standard methods through diphenylamine and orcinol reactions respectively. RESULTS: arsenic trioxide injection produced some pathological conditions, drastic changes (mainly reduction of protein bands) in protein sub-fractions, reduced DNA and RNA contents in both liver and testis; Ars. Alb-30-fed arsenic-intoxicated mice showed revival and restoration in both liver and testis as revealed by gel patterns and quantitative assay of DNA and RNA. CONCLUSION: Efficacy of the homeopathic drug Ars. Alb-30 in reducing arsenic-induced damage to protein and nucleic acids is substantiated and the mechanism of action of the homeopathic drug through expression of regulatory genes inferred.

Analysis of Variance↗

PubMed2006

MALDI-TOF serum protein profiling for the detection of breast cancer.

BACKGROUND: Proteomic expression profiling has been suggested as a potential tool for the early diagnosis of cancer and other diseases. The objective of our study was to assess the feasibility of this approach for the detection of breast cancer. MATERIALS AND METHODS: In a randomized block design pre-operative serum samples obtained from 78 breast cancer patients and 29 controls were used to generate high-resolution MALDI-TOF protein profiles. The spectra generated using C8 magnetic beads assisted mass spectrometry were smoothed, binned and normalized after baseline correction. Linear discriminant analysis with double cross-validation, based on principal component analysis, was used to classify the protein profiles. RESULTS: A total recognition rate of 99%, a sensitivity of 100%, and a specificity of 97.0% for the detection of breast cancer were shown. The area under the curve of the classifier was 98.3%, which demonstrates the separation power of the classifier. The first 2 principal components account for most of the between- group separation. CONCLUSIONS: Double cross-validation showed that classification could be attributed to actual information in the protein profiles rather than to chance. Although preliminary, the high sensitivity and specificity indicate the potential usefulness of serum protein profiles for the detection of breast cancer.

Adult↗

PubMed1976

Further characterization of rat liver mitochondrial fractions. Lipid composition and synthesis, and protein profiles.

1. Heavy and light mitochondrial fractions obtained by differential centrifugation were further characterized with respect to their lipid composition and synthesis and protein profiles, as seen by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 2. The light mitochondrial fraction was rich in total lipids, phospholipids and cholesterol. The cardiolipin content, however, was low. 3. Rates of [3H]glycerol incorporation into phospholipids of heavy mitochondria and microsomal fractions were almost identical. On the other hand, incorporation into the individual phospholipids in light mitochondria was about 4-6 times higher. Incorporation into cardiolipin of light mitochondria was about 10-fold higher than in the heavy mitochondria. 4. Analysis of protein profiles by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis showed that the pattern obtained for the light mitochondria was similar to that for heavy mitochondria. However, the light fraction was relatively poor in high-molecular-weight proteins and rich in low-molecular-weight proteins. The microsomal protein profile was altogether different. 5. The significance of these findings is discussed in relation to mitochondrial biogenesis.

Animals↗

PubMed1998

Mother's milk protein profile, a possible biomarker for human exposure to persistent insecticides.

The electrophoretic protein profile and residue levels of selected persistent insecticides were investigated in 160 mother's milk samples representing 20 different locations in Egypt. Nine major protein bands were detected in all of the samples. These protein bands were designated as lactoferrin, albumin, SIgA heavy chain, casein I, casein II, SIgA light chain, casein III, lysozyme and alpha-lactalbumin. Residue levels of DDT and its metabolites as well as lindane and its other hexachlorocyclohexane isomers were determined using electron capture gas chromatography and confirmed by gas chromatography/mass spectrometric analysis. Samples containing relatively higher residue levels of the DDT group (DDT, DDE and DDD) showed significant effects on the levels of lysozyme and alpha-lactalbumin bands relative to samples with low or no residue levels. On the other hand, the casein subunits were mostly affected by the residue levels of hexachlorocyclohexane isomers (alpha, beta, gamma and delta isomers). The two patterns showed characteristic dose response correlation suggesting that the protein profile of human milk may serve as a quick biomarker for exposure to persistent insecticides.

Albumins↗

PubMed2001

Protein profiles of the Chinese hamster ovary cells in the resting and proliferating stages.

Identification and characterization of the proteins that regulate the transition from the resting stage (G0) through G1 to S phase of the cell cycle are of central importance to understand the control of cell proliferation and chromosome replication. Unlike in lower organisms, where relatively small numbers of key factors are involved in this process, the factors involved in the same control mechanisms in mammalian systems are much more complex. Furthermore, accumulating lines of evidence now suggest that the nuclear matrix and chromatin organization also play an essential role for the cell cycle control in mammalian cells. To gain a better understanding of the overall dynamics and changes of the protein factors in the context of matrix/chromatin organization, we examined the protein profiles of the Chinese hamster ovary (CHO) cells in different cell cycle compartments. The methods used in this study included subcellular fractionations (cytosol, nuclear extraction, chromatin, and nuclear matrix), two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), silver staining, and immunoblotting. As expected, significant changes of protein profiles were observed when cells entered into proliferating stages from G0. Among approximately 1200 protein spots analyzed by 2-D PAGE, at least 12 showed marked increase or decrease at this transitional period. Further cell-cycle progression from G1 to S phase showed less dramatic changes of overall protein protile. However, the profile of certain proteins showed rather dramatic changes of their subcellular localization during this transitional period. In particular, the levels of proliferating cell nuclear antigen (PCNA) in the nuclear matrix and chromatin dramatically increased in mid-G1 and in the beginning of S phase, respectively, while the overall PCNA level was relatively constant throughout the cell cycle.

Animals↗