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At least 541 records · Page 30Linked to original sources
PubMed2002

Tritrophic interactions between cabbage cultivars with different resistance and fertilizer levels, cruciferous aphids and parasitoids under field conditions.

Tritrophic interactions involving cabbage Brassica oleracea var. capitata cultivars +/- fertilizer, Brevicoryne brassicae (Linnaeus) and Myzus persicae (Sulzer), and the parasitoids Diaeretiella rapae (M'cIntosh) and Aphidius sp. were conducted in 1998 and 1999. Brevicorne brassicae was the dominant aphid species on all cultivars +/- fertilizer, except for some treatments in late season 1998. Ruby Ball (red-leaved with antixenosis factors for B. brassicae alates) +/- fertilizer was consistently less colonized by aphids in early stages of plant growth, although only significantly so compared with Derby Day (green-leaved, susceptible to aphids) without fertilizer for B. brassicae and Minicole (green-leaved with antibiosis factors for B. brassicae) with fertilizer for M. persicae. In early 1999, only B. brassicae was present and no significant differences between cultivars were seen. In the mid to late season 1998, the highest aphid infestations were usually found on Derby Day, although only significantly so for B. brassicae, in some treatments. In 1999, higher aphid infestations were observed on Derby Day in mid to late season and some significant differences were found for M. persicae as well as for B. brassicae. In both years, Ruby Ball had the greatest mummy:aphid ratios early season, with no consistent difference between the other cultivars. Later in the season, mummy:aphid ratios were generally highest on Minicole. Parasitism differed in seasonal occurrence and relative abundance. Diaeretiella rapae mummies were found earlier than Aphidius sp. There was evidence of a beneficial interaction between the degree of plant resistance and biological control in early to mid season.

Animals↗
PubMedDate not supplied

Maximal horizontal flight performance of hummingbirds: effects of body mass and molt.

Hovering and fast forward flapping represent two strenuous types of flight that differ in aerodynamic power requirement. Maximal capabilities of ruby-throated hummingbirds (Archilochus colubris) in hovering and forward flight were compared under varying body mass and wing area. The capability to hover in low-density gas mixtures was adversely affected by body mass, whereas the capability to fly in a wind tunnel did not show any adverse mass effect. Molting birds that lost primary flight feathers and reduced wing area also displayed mass loss and loss of aerodynamic power and flight speed. This suggests that maximal flight speed is insensitive to short-term perturbations of body mass but that molting is stressful and reduces the birds' speed and capacity for chase and escape. Hummingbirds' flight behavior in confined space was also investigated. Birds reduced their speeds flying through a narrow tube to approximately one-fifth of that in the wind tunnel and did not display differences under varying body mass and wing area. Hence, performance in the flight tube was submaximal and did not correlate with performance variation in the wind tunnel. For ruby-throated hummingbirds, both maximal mass-specific aerodynamic power derived from hovering performance in low-density air media and maximal flight velocity measured in the wind tunnel were invariant with body mass.

Animals↗
PubMed1993

Optical imaging of shock waves produced by a high-energy electromagnetic transducer.

An electromagnetic acoustic transducer (EMAT) and acoustic lens have been constructed to provide reproducible and well characterized acoustic shock waves in water. The peak pressure amplitude variation for the unfocused and focused shock waves was found to be within +/- 5%. The focal region has been characterized both optically and by conventional piezoelectric pressure measurements. Optical investigations centre around ruby laser illuminated Schlieren and focused shadow imaging. The temporal resolution of the optical technique was determined by the ruby laser pulse length of 20 ns, giving a spatial resolution of 30 microns for a shock front in water.

Acoustics↗
PubMed1992

Lasers in neurosurgery: a historical overview.

This contribution to the history of laser applications to neurosurgical patients gives the background against which the subsequent developments took place. It covers the important facts regarding the theoretical formulations that led to the invention of the world's first laser in 1960. This was a pulsed ruby laser, which proved lacking in desirable surgical procedures, and at high powers was shown to be damaging to vital organs, such as the brain. It could be lethal to small animals. These very early tests of laser tissue and organ interaction included studies on protein in solution, cultured cells, brain, spinal cord and their surrounding tissues, and transplantable melanomas, as well as ependymoblastomas in mice. Fortunately, the continuous wave CO 2 laser came along in 1967 to replace both the pulsed ruby and neodymium-in-glass lasers. The CO 2 laser was quickly seen to possess surgical properties, namely, vaporization, cutting, hemostasis, and sterilization, without additional damage at a distance or remotely in time. Research studies on normal and pathologic tissues in and around the brain and spinal cord quickly and dramatically showed the potential for benefit to animal and human patients with experimental and naturally occurring neoplasms, burns, and decubitus ulcers. Lasers in neurosurgery are used largely for benign and malignant brain and spinal neoplasms. For benign tumors, debulking of tumor mass, and ablation of unwanted neoplasm without damaging adjacent, vital, functioning neural tissue, the laser adds another therapeutic adjunct and will, at times, aid in complete tumor removal. For malignant vascular growths, the laser will aid greatly in the safest possible excision with maximum hemostasis. Studies around the world, ongoing or planned, will surely extend the uses of lasers for neurosurgery into vascular, infectious, and reconstructive fields, particularly with the proliferation of laser instruments that exploit additional wavelengths into the larger infrared levels and even the shorter ultraviolet region. Experimental and, to some extent, clinical applications are testing the value of Nd:YAG, holmium:YAG, and erbium:YAG lasers in various surgical fields.

History, 20th Century↗
PubMed2002

Effect of dietary sorghum cultivars on the storage stability of broiler breast and thigh meat.

A total of 450 1-d-old male broiler chicks were fed a corn-soy-flax meal-based diet (control), two cultivars of sorghum--Ruby Red (low tannin content) and Valpo Red (high tannin content)--were used at 10% level. Birds were slaughtered at the end of 42-d feeding trial. Boneless, skinless breast and thigh muscles were separated and ground to make patties. Half of the breast and thigh meat patties were individually packaged in zipper bags, and 2-TBA-reactive substances (TBARS) and colors of the patties were determined after 0 and 7 d of storage at 4 C. The other half was cooked and vacuum-packaged. The vacuum-packaged patties were used to determine time-dependent volatile production and oxidative change during 12-h holding time before analyses. Thigh meat from broilers fed the Valpo Red cultivar produced lower TBARS than that from control at Day 0. The amounts of aldehydes and sulfur compounds of cooked breast meats were lower from chickens fed the Valpo Red cultivar than those fed the control or Ruby Red cultivar. Dietary Valpo Red cultivar improved the oxidative stability of breast meat 8 and 12 h after cooking. Dietary sorghum slightly improved the color a* stability of raw thigh meat patties. This result indicated that feeding sorghum to broilers could improve some measures of the storage stability of broiler meat, but sorghum with high tannin content was more effective than that with low tannin content.

Aldehydes↗
PubMed2002

Muscle fatty acid composition and thiobarbituric acid-reactive substances of broilers fed different cultivars of sorghum.

Two experiments were conducted to evaluate the production parameters, muscle fatty acid composition, and lipid stability of birds fed four different cultivars of sorghum. Experiment 1 used ruby red and valpo red, and Experiment 2 used kinsman and mason cultivars of sorghum. The basal diet was a corn, soy, and flax meal, and sorghum cultivars were substituted at 10 or 5% in Experiments 1 and 2. No significant difference was observed for BW, feed consumption or feed efficiency (P > 0.05). The total lipids were reduced (P < 0.05) in the leg muscle of broilers fed the mason cultivar. The TBA-reactive substances (TBARS) were lower (P < 0.05) in the leg muscle of birds fed the mason cultivar when compared to kinsman or control diet. The content of n-3 polyunsaturated fatty acids were higher (P < 0.05) in the breast and leg muscles of birds fed ruby red, kinsman, and mason cultivars of sorghum.

Animal Feed↗
PubMed1993

Proposal for rejection of Agrobacterium tumefaciens and revised descriptions for the genus Agrobacterium and for Agrobacterium radiobacter and Agrobacterium rhizogenes.

The 16S rRNA sequences of seven representative Agrobacterium strains, eight representative Rhizobium strains, and the type strains of Azorhizobium caulinodans and Bradyrhizobium japonicum were determined. These strains included the type strains of Agrobacterium tumefaciens, Agrobacterium rhizogenes, Agrobacterium radiobacter, Agrobacterium vitis, Agrobacterium rubi, Rhizobium fredii, Rhizobium galegae, Rhizobium huakuii, Rhizobium leguminosarum, Rhizobium loti, Rhizobium meliloti, and Rhizobium tropici. A phylogenetic analysis showed that the 15 strains of Agrobacterium and Rhizobium species formed a compact phylogenetic cluster clearly separated from the other members of the alpha subclass of the Proteobacteria. However, Agrobacterium species and Rhizobium species are phylogenetically entwined with one another, and the two genera cannot be separated. In the Agrobacterium species, the strains of biovar 1, biovar 2, Agrobacterium rubi, and Agrobacterium vitis were clearly separated. The two biovars exhibited homogeneity in their phenotypic, chemotaxonomic, and phylogenetic characteristics, and two species should be established for the two biovars. We considered the nomenclature of the two biovars, and revised descriptions of Agrobacterium radiobacter (for the biovar 1 strains) and Agrobacterium rhizogenes (for the biovar 2 strains) are proposed. The name Agrobacterium tumefaciens is rejected because the type strain of this species was assigned to Agrobacterium radiobacter, and consequently the description of the genus Agrobacterium is revised.

Agrobacterium tumefaciens↗
PubMed2004

Isolation of a dehydrin cDNA from orange and grapefruit citrus fruit that is specifically induced by the combination of heat followed by chilling temperatures.

Dehydrins (DHNs; late embryogenesis abundant D-11) are a family of plant proteins induced in response to environmental stresses such as water stress, salinity and freezing or which occur during the late stages of embryogenesis. Previously, it was reported that citrus contains a small gene family encoding a unique class of dehydrins that differs from most other plant dehydrins in various respects, such as having an unusual K-segment similar to that of gymnosperms. In the present study, we identified by cDNA differential display analysis a 'Navel' orange 202-bp polymerase chain reaction (PCR) fragment, which encoded the typical plant angiosperm-type K-segment consensus sequence, and of which the expression was down-regulated by exposure to low oxygen levels. The full-length cDNA sequence of the orange DHN, designated csDHN (for Citrus sinensis DHN), was further isolated by 5'-and 3'-RACE; it had a total length of 933 bp and encoded a predicted polypeptide of 235 amino acids. In addition, the same 202-bp 'Navel' dehydrin PCR fragment was used to screen a 'Star Ruby' grapefruit flavedo cDNA library, and its full-length grapefruit homologue, designated cpDHN (for C. paradisi DHN) was isolated and found to have a total length of 1024 bp and to encode a predicted polypeptide of 234 amino acids. The defined orange and grapefruit DHN proteins were completely identical in the 196 amino acids of their N-terminus but differed in their C-terminus region. Overall, the csDHN and cpDHN proteins share 84% identity and contain the conserved dehydrin serine cluster (S-segment) and a putative nuclear localization signal, but csDHN has one conserved dehydrin K-segment consensus sequence, whereas cpDHN contains two dehydrin K-segments. Both csDHN and cpDHN represent single copy genes, in 'Navel' orange and 'Star Ruby' grapefruit genomes, respectively. We found that the cpDHN gene was consistently expressed in the fruit peel tissue at harvest, but that its message levels dramatically decreased during storage at either ambient or low temperatures. However, a pre-storage hot water treatment, given to enhance fruit-chilling tolerance, increased cpDHN mRNA levels during the first 3 weeks of cold storage at 2 degrees C, and enabled the message levels to be retained for up to a further 8 weeks of cold storage at 2 degrees C. The hot water treatment by itself had no inductive effect on cpDHN gene expression when the fruits were held at non-chilling temperatures. Other stresses applied to the fruit, such as wounding, UV irradiation, water stress, low oxygen and exposure to the stress hormone ethylene decreased DHN mRNA levels, whereas abscisic acid had no effect at all.

Journal Article↗
PubMed1977

Modification of pigmentation patterns in allophenic mice by the W gene.

Mouse embryos heterozygous at the W locus were combined with embryos which were wild type at this locus but homozygous for albino. The resulting allophenics displayed an unusual pigmentation phenotype consisting of entirely white fur and ruby-coloured eyes. Microscopic examination showed the eye pigment to be located exclusively in the retinal epithelium, which was a mosaic of black and white sectors. This ruby-eyed white pattern corresponds to what would have been expected for WWCC in equilibrium wwcc mosaics but not for WwCC in equlibrium wwcc mice. WW mice are black-eyed whites, but Ww mice have black eyes and black fur, except for a small ventral white spot. These results suggest that melanocytes of the Ww genotype, although capable of producting normally pigmented fur in Ww animals, fail to populate hair follicles when the competition with wwcc (albino) melanocytes that are wild type at the W locus. The genotype of these WwCC in equilibrium wwcc alophenes was proved by progeny testing. This is apparently the first report of a single gene change affecting the competitive ability of cells in allophenic mice, and suggests that such changes may play a significant role in the clonal selection of embryonic cells during development.

Animals↗
PubMed1975

Carbohydrate catabolism of selected strains in the genus Agrobacterium.

Radiorespirometric and enzyme analyses were used to reveal the glucose-catabolizing mechanisms functioning in single strains of seven presumed Agrobacterium species. The Entner-Doudoroff and pentose cycle pathways functioned in A. radiobacter, A. tumefaciens, A. rubi, and A. rhizogenes. Whereas both catabolic pathways were utilized to an almost equal degree in the A. radiobacter and A. tumefaciens strains, use of the Entner-Doudoroff pathway predominated in the A. rubi and A. rhizogenes strains. A stellulatum catabolized glucose almost solely through the Entner-Doudoroff pathway. In A. pseudotsugae and A. gypsophilae, glucose was metabolized mainly through the Emden-Meyerhof-Parnas pathway; the pentose phosphate pathway was also utilized.

Aldehyde-Lyases↗
PubMed1967

Deoxyribonucleic acid homology and taxonomy of Agrobacterium, Rhizobium, and Chromobacterium.

Hybridization experiments were carried out between high molecular weight, denatured, agar-embedded deoxyribonucleic acid (DNA) and homologous, nonembedded, sheared, denatured (14)C-labeled DNA from a strain of Agrobacterium tumefaciens and Rhizobium leguminosarum (the reference strains) in the presence of sheared, nonembedded, nonlabeled DNA (competing DNA) from the same or different nomen-species of Agrobacterium, Rhizobium, Chromobacterium, and several other organisms. Percentage of DNA homology was calculated from the results. The findings are discussed in relation to previous taximetric studies, present classification schemes, and guanine-cytosine content of the DNA. Strains of A. tumefaciens, A. radiobacter, A. rubi, A. rhizogenes, R. leguminosarum, and R. meliloti exhibited a mean percentage of DNA homology greater than 50 with the two reference strains. A. tumefaciens, A. radiobacter, and A. rubi were indistinguishable on the basis of DNA homology, with strain variations for this group involving up to 30% of their base sequences. The remainder of the organisms studied fall into at least six distinct genetic groups: (i) R. (Agrobacterium) rhizogenes, which is more homologous to R. leguminosarum than to the A. tumefaciens-A. radiobacter group; (ii) R. leguminosarum; (iii) R. meliloti; (iv) R. japonicum, which has a mean DNA homology of some 38 to 45% with the reference strains; (v) Chromobacterium, which is as genetically remote from the reference strains as, for example, Pseudomonas; and (vi) A. pseudotsugae strain 180, which has a DNA homology with A. tumefaciens and R. leguminosarum of only about 10%. Since this latter homology value is similar to what was found after hybridizations between the reference strains and organisms such as Escherichia coli and Bacillus subtilis, A. pseudotsugae should definitely be removed from the genus.

Bacillus subtilis↗
PubMed1983

Laser-induced thrombi in rat mesenteric vessels and antithrombotic drugs.

A method to induce microthrombi in small mesenteric arteries (phi 15-25 microns) has been developed to study platelet reactions and to investigate antithrombotic drugs. A high power water immersion interference contrast system, based on a Leitz Orthoplan microscope, was used. In Nembutal-anesthetized male Wistar rats and in fawn hooded bleeder rats, vascular lesions were produced with a Hadron-512-Ruby-bio-laser or with a Coherent CR-2 supergraphite ion laser (Argon laser). The ruby laser produced intravascular precipitates consisting of proteins and erythrocytes which usually stuck to the vessel wall and on which platelets immediately adhered, transformed with pseudopode formation and formed loose aggregates. The Argon laser induced vascular damage, usually without intravascular heat precipitates which also led to platelet adhesion, transformation and aggregation. Fibrin threads rarely formed in the early platelet thrombi. Thrombus formation was significantly reduced in this model by dipyridamole (10 mg/kg orally), by Ditazole (50 mg/kg orally), heparin (100 IU/kg i.v.), Molsidomine (50 mg/kg orally), Nafazatrom (Bay G 6575, 10 mg/kg i.v. and orally), Ticlopidine (100 and 200 mg/kg orally) and Tioxaprofen (EMD 26644, 10 mg/kg orally). Thrombus formation was also significantly reduced in fawn hooded bleeder rats.

Animals↗
PubMed1997

Transient hovering performance of hummingbirds under conditions of maximal loading.

Maximal load-lifting capacities of six ruby-throated hummingbirds (Archilochus colubris) were determined under conditions of burst performance. Mechanical power output under maximal loading was then compared with maximal hovering performance in hypodense gas mixtures of normodense air and heliox. The maximal load lifted was similar at air temperatures of 5 and 25 degrees C, and averaged 80% of body mass. The duration of load-lifting was brief, of the order of 1 s, and was probably sustained via phosphagen substrates. Under maximal loading, estimates of muscle mass-specific mechanical power output assuming perfect elastic energy storage averaged 206 W kg-1, compared with 94 W kg-1 during free hovering without loading. Under conditions of limiting performance in hypodense mixtures, maximal mechanical power output was much lower (131 W kg-1, five birds) but was sustained for longer (4 s), demonstrating an inverse relationship between the magnitude and duration of maximum power output. In free hovering flight, stroke amplitude and wingbeat frequency varied in inverse proportion between 5 and 25 degrees C, suggesting thermoregulatory contributions by the flight muscles. Stroke amplitude under conditions of maximal loading reached a geometrical limit at slightly greater than 180 degrees. Previous studies of maximum performance in flying animals have estimated mechanical power output using a simplified actuator disk model without a detailed knowledge of wingbeat frequency and stroke amplitude. The present load-lifting results, together with actuator disc estimates of induced power derived from hypodense heliox experiments, are congruent with previous load-lifting studies of maximum flight performance. For ruby-throated hummingbirds, the inclusion of wingbeat frequency and stroke amplitude in a more detailed aerodynamic model of hovering yields values of mechanical power output 34% higher than previous estimates. More generally, the study of performance limits in flying animals necessitates careful specification of behavioral context as well as quantitative determination of wing and body kinematics.

Animals↗
PubMed2006

Variations in content of active ingredients causing drug interactions in grapefruit juice products sold in California.

The content of the active ingredients of grapefruit juice, naringin, naringenin and bergapten, was determined in 20 different commercial products of grapefruit juice sold in California. These included Minute Maid, Dole, Tropicana, Ocean Spray, Ralps, Albertson, Stater Bros, Vons, Langers, etc. The concentrations of naringin, naringenin and bergapten in grapefruit juice were assayed by specific HPLC methods. Naringin was found to be the most abundant flavonoid in grapefruit juice products, followed by naringenin and bergapten. The content of naringin varied among the products, ranging from 104 mg/l (Tropicana ruby red) to 628 mg/l (Ralphs white frozen concentrate). The mean contents of naringin in ruby red (158 +/- 66 [SD] mg/l) and pink (279 +/- 123 mg/l) grapefruit juice products were significantly lower than white (481 +/- 94 mg/l) (p <0.005) grapefruit juice products. Content of naringenin also varied from brand to brand and ranged from 3.9 mg/l (Vons white frozen concentrate) to 31.2 mg/l (Tree Sweet pink). Bergapten content was very low in grapefruit juice products ranging from 0 (not detectable) to 5.5 mg/l. There were no significant differences in naringenin and bergapten contents among the three types of grapefruit juice products. The information gained from this study would be useful in predicting the likelihood of grapefruit juice-drug interactions.

5-Methoxypsoralen↗
PubMed1999

Rapid healing of gingival incisions by the helium-neon diode laser.

Fifty-eight extraction patients had one of two gingival flap incisions lased with a 1.4 mw helium-neon (670 nm) diode laser for 30 seconds (fluence = 0.34 J/cm2). Healing rates were evaluated clinically and photographically. Sixty-nine percent of the irradiated incisions healed faster than the control incisions. No significant difference in healing was noted when patients were compared by age, gender, race, and anatomic location of the incision. This study concludes that helium-neon diode lasers, at the previously mentioned energy level, increase the rate of gingival wound healing in 69 percent of patients, without any side effects. For the last 30 years, low-power lasers in dentistry have appeared to stimulate healing rates and increase the rate of repair of injured tissue. Helium-neon and similar lasers emit light in the red (600-700 nm) spectrums and produce energy densities (fluences) below 20 Joules/cm2. They have been studied in a variety of animal tissue culture and human evaluations to determine their ability to increase the rates of wound healing by biostimulation. Over the last three decades, researchers have found that ruby and gas helium-neon (low-power laser radiation) have a biostimulatory effect on living tissue. Studies show that under specific conditions, red spectrum laser light speeds the healing of wounds. Photons from the red light lasers, which include ruby lasers (694 nm), helium-neon gas lasers (632 nm), and helium-neon diode lasers (650-670 nm), appear to stimulate rapid epithelialization and fibroblast (collagen) proliferation in animal and human tissue cultures. Low-power lasers have been reported to reduce post-extraction pain and swelling and to increase rates of wound healing (including scar formation, phagocytosis) in cell culture, animal, and human clinical studies. The new, compact, and inexpensive (under $50) helium-neon diode lasers have produced similar effects. These FDA Class IIIa lasers have no hazards associated with them, although one should avoid direct exposure to the eye for a prolonged period of time. In the past, many biostimulation studies using red spectrum lasers produced confusing data and conflicting results. Some studies reported that the biostimulation effect did not occur in all cases of laser irradiation, while other research reported that it did. Results seem to depend on the delivery of appropriate energy fluence levels (between 1 and 20 J/cm2) and the type of laser (wavelength) used. Several of these studies never described the levels of laser energy used to promote the described biostimulatory results. This caused controversy when determining whether or not helium-neon lasers influence wound healing. Studies suggest that low-power laser exposure can significantly increase the healing rate during the first few days of the healing process; however, studies do not show appreciable net benefit as compared to controls toward the end of a two-week wound repair cycle. The increased healing effect appears to be centered around the early, most sensitive stages of the healing process. Several studies showed optimum tissue healing rates at helium-neon laser exposure levels between 1 J/cm2 and 20 J/cm2. Laser-enhanced biostimulation has been reported to produce metabolic changes within the cells. This results in faster cell division, rapid matrix production (increased collagen, myofibroblasts, etc.), and cell movement. There have been few controlled studies using adequate numbers of human subjects in identifying the beneficial effects of helium-neon laser biostimulation. Ethical concerns, bulky equipment, and problems with biased study designs have frustrated a practical evaluation of laser biostimulation for general dental practice. A recently published "preliminary" study involving 52 patients was designed to reduce these issues. The purpose of this study is to complement the above research and to evaluate whether helium-neon diode laser radiation at average fluences of 0.34 J/c

Adult↗
PubMed1983

Enhancement of lettuce yield by manipulation of light and nitrogen nutrition.

Several levels of photosynthetic photon flux density (PPFD) were tested for effects on growth of 4 cultivars of lettuce (Lactuca sativa L.) under controlled-environment conditions. Growth of 'Salad Bowl', 'Bibb', and 'Ruby' was greater at 932 micromoles s-1 m-2 than at < or = 644 micromoles s-1 m-2 under a 16-hour photoperiod. Thirty mM NO3- or 5 mM NH4+ + 25 mM NO3- increased leaf dry weight while reducing leaf chlorosis in 'Salad Bowl' and 'Grand Rapids' relative to that with 15 mM NO3-, and reduced leaf purpling in 'Bibb' and 'Ruby' with little or no effect on yield. Continuous illumination with 455 or 918 micromoles s-1 m-2 stimulated yield of 'Salad Bowl' and 'Bibb' when 30 mM N as NH4+ + NO3- was used relative to that with 15 mM NO3-.

Culture Media↗
PubMed2002

Proteomic analysis and identification of new biomarkers and therapeutic targets for invasive ovarian cancer.

Epithelial ovarian cancer kills almost 16 000 women each year in part due to late stage of presentation and lack of reliable biomarkers for disease detection. CA-125, the currently accepted serum marker, alone lacks the sensitivity for early stage diagnosis, as only 50% of early stage cases are detected with this marker. Although more early stage cases may be detected by lysophosphatidic acid, this marker is also elevated in other cancers. One major objective of the NCI-FDA Tissue Proteomics Initiative has been to combine the technique of laser capture microdissection (LCM) of epithelial tumor cells in human tissue specimens with two-dimensional gel electrophoresis (2-D PAGE) to identify proteins that may serve as invasive ovarian cancer-specific biomarkers for early detection and/or new therapeutic targets. We performed 2-D PAGE on lysates from five microdissected ovarian tumors (three invasive ovarian cancers and two noninvasive, low malignant potential (LMP) ovarian tumors). We then compared silver stained 2-D gels created from microdissected lysates with SYPRO-Ruby stained 2-D PAGE profiles of the patient-matched undissected bulk tumor lysates from all five patients. Twenty-three proteins were consistently differentially expressed between both the LMP and three invasive ovarian tumors in the limited study set. Thirteen were uniquely present in all three of the invasive ovarian cancer cases and absent or underexpressed in the two LMP cases. Ten were uniquely present in the LMP cases but absent or underexpressed in all invasive ovarian cancer cases. Credentialing and preliminary target validation of the mass spectrometry identified proteins cut from the Ruby-red stained gels was performed by LCM coupled Western blot and reverse-phase array technology in a study set of six cases (the aforementioned five cases used in the 2-D PAGE profiling component of the study plus one additional LMP case). The analysis revealed that the 52 kDa FK506 binding protein, Rho G-protein dissociation inhibitor (RhoGDI), and glyoxalase I are found to be uniquely overexpressed in invasive human ovarian cancer when compared to the LMP form of this cancer. The direct comparison of LCM generated proteomic profiles of invasive vs. LMP ovarian cancer may more directly generate important markers for early detection and/or therapeutic targets unique to the invasive phenotype.

Biomarkers, Tumor↗
PubMed2002

Development of improved cell lysis, solubilization and imaging approaches for proteomic analyses.

Analysis of complex biochemical processes at the level of the proteome requires methods that quantitatively solubilize cytosolic and membrane bound proteins yet are compatible with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In addition, it is often necessary to employ several highly sensitive detection methods to identify key proteins that are modified or exhibit a change in expression levels in response to a given experimental stimulus or condition. Methods were developed that efficiently extract tissues or lyse cultured cells and quantitatively solubilize proteins in a single step without the need to shear nucleic acids. These approaches utilize urea, thiourea, a mixture of detergents, low levels of an ampholyte blend, reductant and a combination of alcohols. To aid in the detection of low abundance proteins and the accurate identification of specific proteins of interest in these samples, two approaches were pursued. In one, proteins are transferred from two-dimensional (2-D) gels to blot membranes. Proteins are then detected by staining with SYPRO Ruby and the resulting 2-D protein pattern is captured using a charge-coupled device (CCD) camera. The blots are then probed with antibodies directed against the protein(s) or functionalities of interest. The resulting chemiluminescent blot image is also generated with the CCD camera and the fluorescent SYPRO Ruby image is recaptured again without moving the membrane. It is thereby possible to generate a direct image overlay of the blot pattern on that of the stained protein pattern. This approach significantly aids in the accurate identification of the dye-stained protein that is detected by the specific antibody. In addition to detecting protein post-gel transfer, a second approach utilizes protein samples labeled with fluorescent dyes prior to 2-D electrophoresis in an effort to increase the sensitivity of protein detection and to facilitate protein quantitation. It is also possible to stain the blots with different dyes and overlay these images as well. Using these approaches, it is possible to perform more rapid and accurate comparative analyses and proteomic, post-gel characterization of proteins of interest than using comparative image analysis of multiple gels.

Blotting, Western↗