The proteoglycans (PGs) in the guinea pig seminalvesicle were demonstrated ultrastructurally by both cuprolinic blue (CB) and ruthenium red (RR) staining. The PGs appeared as electron-dense granules...Full Text Available
Between October 1990 and March 1991, 8 patients with cystic lesions in the region of the posterior bladder wall, the seminalvesicles and the prostate were examined by MRI. In all cases there was accurate characterisation of the lesion and of its anatomy. In 3 patients with abnormalities in the retrovesical space there was very good agreement with the operative findings. 4 patients were treated conservatively. In these patients there was good agreement with the findings on cystoscopy, CT and sonography. (orig.).
The effects of oral administration of aqueous extract of Bulbine natalensis Baker stem at daily doses of 25, 50, and 100mg/kg body weight on the reproductive function of Wistar rats were evaluated. The indices of mating and fertility success as well as quantal frequency increased after 7 days of treatment in all the dose groups except the 100mg/kg body weight group. The number of litters was not statistically different (P>0.05) from the control. Whereas the absolute weights of the epididymis, seminalvesicle, and prostate were not affected, that of the testes was significantly increased. The epididymal sperm count, motility, morphology, and viscosity were not different from the control after 7 days of treatment. The male rat serum testosterone, progesterone, luteinizing hormone, and follic...
Summary During a histopathological survey of the geoduck, Panopea abbreviata (Hiatellidae), and the razor clam, Ensis macha (Pharidae), in northern Patagonian gulfs (Argentina), turbellarian worms were found in the intestine lumen. In this work, we described a new species of Paravortex (Platyhelminthes, Graffillidae). We collected the hosts by scuba diving and dissected alive for studying the turbellarians. In this new species, the pharynx continues in a short esophagus and the latter in a saccular intestine. Gravid specimens have from 2 to 8 twin embryos in the parenchyma. A short penis papilla arises from the seminalvesicle and the gonopore presents a strong sphincter. This species resembled P. nicolli Szidat, 1965, which was described paraziting Mytilus edulis platensis in Buenos Aires...
Techniques of chemical analysis, amino acid sequencing and autoradiography are being used to study the frequency of incorporation of normally noncoded amino acids into hemoglobins and seminal fluid proteins. We are studying, by the sequencing of radiolabeled proteins followed by the recovery of ["3H] isoleucine phenylthiohydantoin by high-performance liquid chromatography, the frequency at which normally noncoded isoleucine is incorporated into hemoglobin because of base-substitution mutations versus translational errors. Irradiation increases the isoleucine content of human hemoglobin and the frequency of substitution of isoleucine for specific amino acids in rabbit hemoglobin. Studies to date indicate that these techniques have been developed sufficiently for initial analysis of the potential of drugs and environmental pollutants to induce base-substitution mutations in mammalian somatic cells.
Purpose: The results of therapy in 288 men with pathologic Stage C prostate cancer who underwent radical retropubic prostatectomy (RRP) were analyzed to determine the effects of adjuvant therapy. Methods and Materials: Twenty-seven of the 288 patients received preoperative neoadjuvant hormonal therapy (leuprolide acetate). Postoperatively, 60 patients received adjuvant radiotherapy (RT) to the prostate bed. Follow-up ranged from 3 to 83 months (median = 32 months). Freedom from failure (FFF) was defined as maintaining a serum PSA level of #<=# 0.3 ng/ml. Results: The FFF was 61% at 3 years and 45% at 5 years for the entire group. The FFF following RRP plus RT was 75% at 3 years and 57% at 5 years as compared to 56% at 3 years and 40% at 5 years for RRP without RT (p = 0.049). The FFF following RRP plus neoadjuvant hormonal therapy was 58% at 3 years and 40% at 5 years as compared to 60% at 3 years and 45% at 5 years following RRP without hormonal therapy (p = 0.3). In patients ...
Background and purpose: We evaluate the feasibility of concomitant and adjuvant docetaxel combined with three-dimensional conformal radiotherapy (3D-CRT) and androgen deprivation in high-risk prostate carcinomas. Methods: Fifty men with high-risk localized prostate cancer (16), locally advanced (28) or very high-risk prostate cancer (6) were included. Seventy Gy were delivered on prostate and seminalvesicles in 35 fractions, concurrently with weekly docetaxel (20 mg/m"2). Three weeks after the completion of 3D-CRT, docetaxel was given for 3 cycles (60 mg/m"2), every 3 weeks. Patients had to receive LHRH agonist during 3 years. Results: The intent to treat analysis shows that four patients out of 15 stopped prematurely the chemotherapy due to grade 3-4 acute toxicity. In the per protocol analysis, 46 patients completed a full-dose chemoradiation regimen representing 413 cycles: five patients experienced a grade 3 toxicity, and 15 patients ...
Unlike in normal rabbits, pulmonary rapidly adapting receptors (RARs) in rabbits with chronic mitral regurgitation (MR) do not respond to small changes in extravascular...Full Text Available
Synaptic vesicle endocytosis is critical to maintain synaptic communication during intense stimulation. Here we describe Tweek, a conserved protein that is required for synaptic vesicle recycling....Full Text Available
In the presence of electrochemical energy, several branched-chain neutral and acidic amino acids were found to accumulate in membrane vesicles of Bacillus stearothermophilus. The membrane vesicles contained...Full Text Available
We develop and investigate numerically a thermodynamically consistent model of two-dimensional multicomponent vesicles in an incompressible viscous fluid. The model is derived using an energy...Full Text Available
Phospholipid vesicles were found to be lysed by exposure to free radicals derived from irradiated glycine and galactose. The decomposition yield increased with vesicle concentration, and attained values of 0.0006 vesicles destroyed per radical added. Ionic charge at the phospholipid end groups was found to be more important than degree of unsaturation in determining this yield. Radicals derived from irradiated serine and alanine had no detectable lytic action at the vesicle concentrations tested.
We have developed a new strategy for immunization of wild rabbit populations against myxomatosis and rabbit hemorrhagic disease (RHD) that uses recombinant viruses based on a naturally attenuated field...Full Text Available
The transfer kinetics of the negatively charged glycosphingolipid II"3-N-acetylneuraminosyl-gangliotetraosylceramide (GM_1) were investigated by monitoring tritiated GM_1 movement between donor and acceptor vesicles. After appropriate incubation times at 45 "0C, donor and acceptor vesicles were separated by molecular sieve chromatography. Donors were small unilamellar vesicles produced by sonication, whereas acceptors were large unilamellar vesicles produced by either fusion or ethanol injection. Initial GM_1 transfer to acceptors followed first-order kinetics with a half-time of about 40 h assuming that GM_1 is present in equal mole fractions in the exterior and interior surfaces of the donor vesicle bilayer and that no glycolipid flip-flop occurs. GM_1 net transfer was calculated relative to that of ["1"4C]cholesteryl oleate, which served as a nontransferable marker in the donor ...
Purpose: The purpose of this study was to evaluate the efficacy and toxicity of replication-competent adenovirus-mediated double suicide gene therapy in an adjuvant setting with external beam radiation therapy (EBRT) in an experimental prostate cancer model in preparation for a Phase I clinical study in humans. Methods: For efficacy studies, i.m. DU145 and intraprostatic LNCaP C4-2 tumors were established in immune-deficient mice. Tumors were injected with the lytic, replication-competent Ad5-CD/TKrep adenovirus containing a cytosine deaminase (CD)/herpes simplex virus thymidine kinase (HSV-1 TK) fusion gene. Two days later, mice were administered 1 week of 5-fluorocytosine + ganciclovir (GCV) prodrug therapy and fractionated doses of EBRT (trimodal therapy). Tumor control rate of trimodal therapy was compared to that of EBRT alone. For toxicology studies, immune-competent male mice received a single intraprostatic injection (10"1"0 vp) of the replication-competent Ad5-CD/TKrep ...
To evaluate the relationship between sperm motility and total seminal plasma zinc concentration and high molecular weight zinc bound protein values in infertile Iraqi men. A case-control study was conducted at the Chemistry and Biochemistry Department, College of Medicine, Al-Nahrain University, Baghdad, Iraq between March 2005 to February 2006. The subjects for the study included 60 infertile male patients who were recruited Al-Kadhimiya Teaching Hospital, and Institute of Embryo Research and Infertility Treatment, Baghdad, Iraq. They were categorized according to their seminal parameters to oligozoospermia (n=32), azoospermia (n=22), and asthenozoospermia (n=6). Thirty nine fertile men (age range 31.87 +/- 3.76 years) were selected as controls, whose partners had conceived within the last year before participation with this study, and having normal spermiogram parameters. Seminal plasma zinc concentration and high ...
To evaluate the effect of plugging the biopsy tract in rabbit liver and the pathologic changes caused by plugging materials. Thirty-two New Zealand White rabbits were divided into four groups (eight rabbits in each) and compared with one another. They were labeled group A (control), B (gelfoam), C (fibrin sealant) or D (NBCA). the liver was exposed and biopsied with an 18G disposable biopsy gun. The inner Tru-cut needle was withdrawn and plugging was undertaken through the outer cannula of the biopsy gun. Bleeding times of each material were compared. The rabbits were sacrificed and pathologically evaluated for 17 days.
Amino acid transport in membrane vesicles of Bacillus stearothermophilus was studied. A relatively high concentration of sodium ions is needed for uptake of L-alanine (Kt = 1.0 mM) and L-leucine (Kt...Full Text Available
The rate-limiting step in the transit of dietary fat across the intestinal absorptive cell is its exit from the endoplasmic reticulum (ER) in a specialized ER-to-Golgi transport vesicle, the prechylomicron...Full Text Available
Amino acid transport was studied in membrane vesicles of the thermophilic anaerobic bacterium Clostridium fervidus. Neutral, acidic, and basic as well as aromatic amino acids were transported at 40...Full Text Available
Biomimetic vesicle media which can selectivity extract and concentrate heavy metals (e.g., Pb{sup 2+}) from dilute solutions of 5 ppm and less have been prepared. At a concentration of only 1% 2/v, these non-covalently assembled, unilamellar, surfactant vesicles provide approximately 1,000m{sup 2} of exposed membrane surface area per liter of suspension which gives rise to very rapid rates of metal ion extraction. To facilitate the selective transport of heavy metal ions through the ion impermeable vesicle wall, the vesicles are doped with lipophilic ionophores, both natural and synthetic. In addition, the metal chelating agent, nitrolotriacetate, is encapsulated in the aqueous vesicle core to provide the driving force for metal ion uptake and concentration. Concentration factors in excess of 1000 fold have been observed. Metal-sorbing surfactant vesicles ...
The primary method for neuronal communication involves the extracellular release of small molecules that are packaged in secretory vesicles. We have developed a platform to separate, lyse, and...Full Text Available
The transfer kinetics of the neutral glycosphingolipid gangliotetraosylceramide (asialo-GM1) were investigated by monitoring tritiated asialo-GM1 movement from donor to acceptor vesicles. Two different methods were employed to separate donor and acceptor vesicles at desired time intervals. In one method, a negative charge was imparted to dipalmitoylphosphatidylcholine donor vesicles by including 10 mol% dipalmitoylphosphatidic acid. Donors were separated from neutral dipalmitoylphosphatidylcholine acceptor vesicles by ion-exchange chromatography. In the other method, small, unilamellar donor vesicles and large, unilamellar acceptor vesicles were coincubated at 45 degrees C and then separated at desired time intervals by molecular sieve chromatography. The majority of asialo-GM1 transfer to acceptor vesicles occurred as a slow first-order ...
We present an experimental study of vesicle fusion using light and neutron scattering to monitor fusion events. Vesicles are reproducibly formed with an extrusion procedure using an single amphiphile triethylene glycol mono-n-decyl ether in water. They show long-term stability for temperatures around 20 C, but at temperatures above 26 C we observe an increase in the scattered intensity due to fusion. The system is unusually well suited for the study of basic mechanisms of vesicle fusion. The vesicles are flexible with a bending rigidity of only a few k(H)T. The monolayer spontaneous curvature, Ho, depends strongly on temperature in a known way and is thus tunable. For temperatures where H-0 > 0 vesicles tyre long-term stable, while in the range H-0 < 0 the fusion rate increases the more negative the Spontaneous curvature Through a quantitative;analysis of the fusion rate we ...
The effects of homologous plasma HDL and VHDL fractions on established atherosclerotic lesions were studied in cholesterol-fed rabbits. Atherosclerosis was induced by feeding the animals a 0.5% cholesterol-rich...Full Text Available
The percentages of total airflows over the nasal respiratory and olfactory epithelium of female rabbits were calculated from computational fluid dynamics (CFD) simulations of steady-state inhalation....Full Text Available
The cottontail rabbit papillomavirus (CRPV)/rabbit model has been used to study oncogenicity and immunogenicity of different antigens from the papillomavirus genome and has therefore served...Full Text Available
During the anamnestic response, the sera of four rabbits immunized with a polyvalent pneumococcal vaccine contained large amounts of cryoglobulins belonging to the G and M immunoglobulin classes. These...Full Text Available
The efficacy of beta-lactam antibiotics and amikacin alone and in various combinations against Pseudomonas aeruginosa was studied in a rabbit model simulating a closed-space infection in a locally neutropenic...Full Text Available
The pharmacokinetics of two prodrugs of zidovudine (AZT), 1,4-dihydro-1-methyl-3-[(pyridylcarbonyl)oxy] ester and isoleucinyl ester (DPAZT and IAZT, respectively), were investigated in a rabbit model...Full Text Available
Recent studies have shown Y chromosome microdeletions associated with male infertility. The factors responsible for Y chromosome microdeletions in spermatozoa remain unresolved. However, the environmental pollutants are known to damage DNA in differentiating and maturing germ cells in the male reproductive tract. Therefore, the aim of this study was to investigate the effects of seminal hexachlorocyclohexane (HCH) and its isomers, an environmental pollutant, in 50 fertile and 50 infertile males in relation to semen quality and the incidence of Y chromosome microdeletion in azoospermic factor (AZF) region. As compared to control, an increased HCH level and significantly decreased semen quality were observed in the infertile males. A positive significant association was found between sperm c...
The investigations were conducted at the Center of Biotechnology and Molecular Diagnostics of the All-Russian Animal Husbandry Research Institute, where a number of valuable transgenic animals (rabbits, sheep, and pigs) have been produced over the course of 20 years. The article presents the blood indices of control and transgenic rabbits with an integrated insulin gene.
This work showed the formation of phospholipid vesicle coating on inorganic sericite surface with characterization by combining electron microscopy of FE-SEM, TEM, AFM, and qualitatively evaluated the coated phospholipid vesicle by XPS as a function of etching time. The possibility of phospholipid vesicle mobility on the surface was restrained by the chelation effect of magnesium cation. The stabilization properties of phospholipid vesicles on sericite surface were demonstrated by the various concentration of magnesium cation. The presence of magnesium was found to have a much more pronounced influence on the lipid deposition process. The Mg cation plays an important role for attaching the phospholipids with optimum concentration of 7 mM. Totally, the phospholipid vesicles coating on inorganic powder could be useful for bio-related fields such as cosmetics and drug delivery system ...
The transfer kinetics of the monosiaylated glycosphingolipid, GM_1, between different size phospholipid vesicles was measured using molecular sieve chromatography. At desired time intervals, small unilamellar donor vesicles were separated from large unilamellar acceptor vesicles by elution from a Sephacryl S-500 column ["3H]-GM_1 net transfer was calculated relative to ["1"4C]-cholesteryl oleate, which served as a nontransferable marker in the donor vesicles. The initial GM_1 transfer rate between 1-palmitoyl-2-oleoyl phosphatidylcholine vesicles at 45"0C deviated slightly from first order kinetics and possessed a half time of 3.6 days. This transfer half time is an order of magnitude shorter than that observed from the desiaylated derivative of GM_1. The transfer kinetics are consistent with the authors recent electron microscopic results suggesting a molecular distribution of GM_1 ...
Autophagy is an evolutionarily conserved, catabolic process that involves the entrapment of cytoplasmic components within characteristic vesicles for their delivery to and degradation within lysosomes....Full Text Available
Small unilamellar vesicles which form when gel-state long-chain phosphatidylcholines are mixed with micellar short-chain lecithins undergo an increase in size as the long-chain species melts to its liquid-crystalline form. Analysis of the vesicle population with quasi-elastic light scattering shows that the particle size increases from 90-A radius to greater than 5000-A radius. Resonance energy transfer experiments show total mixing of lipid probes with unlabeled vesicles only when the Tm of the long-chain phosphatidylcholine is exceeded. This implies that the large size change represents a fusion process. Aqueous compartments are also mixed during this transition. 31P NMR analysis of the vesicle mixtures above the phase transition shows a great degree of heterogeneity with large unilamellar particles coexisting with oligo- and multilamellar structures. Upon cooling the vesicles ...
In this work, a mixed cationic--anionic [i.e. cetyltrimethylammonium bromide-sodium octyl sulphate] surfactant system was used to synthesize the magnetite particles. The loading content of iron ions (Fe{sup 3+} and Fe{sup 2+}) into the surfactant system was varied to study its effect on the formation of magnetite, magnetic and morphological properties of the magnetite particles encapsulated by the surfactant vesicles. It has been verified that the vesicle-encapsulated magnetite particles with sizes from 100 to 200 nm were formed, which were almost independent of the loading content of iron ions. However, the morphological structure of the magnetite particles was dependent on the loading content of iron ions and there existed an optimal loading content for a full packing of the surfactant vesicles with the magnetite particles.
In this work, a mixed cationic--anionic [i.e. cetyltrimethylammonium bromide-sodium octyl sulphate] surfactant system was used to synthesize the magnetite particles. The loading content of iron ions (Fe"3"+ and Fe"2"+) into the surfactant system was varied to study its effect on the formation of magnetite, magnetic and morphological properties of the magnetite particles encapsulated by the surfactant vesicles. It has been verified that the vesicle-encapsulated magnetite particles with sizes from 100 to 200 nm were formed, which were almost independent of the loading content of iron ions. However, the morphological structure of the magnetite particles was dependent on the loading content of iron ions and there existed an optimal loading content for a full packing of the surfactant vesicles with the magnetite particles.
Soufriere Hills Volcano had two periods of repetitive Vulcanian activity in 1997. Each explosion discharged the contents of the upper 0.5-2km of the conduit as pyroclastic flows and fallout: frothy pumices from a deep, gas-rich zone, lava and breadcrust bombs from a degassed lava plug, and dense pumices from a transition zone. Vesicles constitute 1-66vol.% of breadcrust bombs and 24-79% of pumices, all those larger than a few tens of m being interconnected. Small vesicles (few hundreds of m) in pumices are interpreted as pre-dating explosion, implying pre-explosive conduit porosities up to 55%. About a sixth of large vesicles in pumices, and all those in breadcrust bombs, are angular voids formed by syn-explosive fracturing of amphibole phenocrysts. An intermediate-sized vesicle population...
The synapsin III gene, SYN3, which belongs to the family of synaptic vesicle-associated proteins, has been implicated in the modulation of neurotransmitter...Full Text Available
The structure of polydispersed populations of unilamellar vesicles is studied by small-angle neutron scattering for three types of lipid systems, namely, single-, two-and four-component vesicular systems. Results of the numerical analysis based on the separated-form-factor model are reported.
The yeast phosphatidylinositol transfer protein (Sec14p) is required for biogenesis of Golgi-derived transport vesicles and cell viability, and this essential Sec14p requirement is abrogated by inactivation...Full Text Available
Amphiphilic pincer palladium complexes bearing hydrophilic and hydrophobic side chains on the planar NCN palladium pincer backbone were designed and prepared via the ligand introduction route. The complexes self-assembled under aqueous conditions to form vesicles with bilayer membranes containing palladium species. The catalytic activity of the vesicles in the Miyaura-Michael reaction in water was investigated. PMID:21837316
Small angle neutron scattering (SANS) on the unilamellar vesicle populations (diameter 500 and 1000 angstrom) was used to characterize lipid vesicles from dimyristoylphosphatidylcholine (DMPC) at three phases (gel, ripple, and liquid). Parameters of vesicle populations and internal structure of the DMPC bilayer were characterized on the basis of the Separated Form Factor (SFF) model. Parameters of the internal bilayer structure (thickness of the membrane and the hydrophobic core, hydration, and surface area of lipid molecule) were determined on the basis of the Hydrophobic-Hydrophilic (HH) approximation of neutron scattering length density across the bilayer r(x) and of the Step Function (SF) approximation of r(x). It was demonstrated in frame of HH approximation that DMPC membrane thickness in liquid phase (T=30 degrees) depends on the membrane curvature. The dependence of the DMPC membrane thickness on temperature was ...
The project involved several studies of vesicles and micelles and their constituent surfactants. The primary objective of the project was an elucidation of factors involved in the stabilization and destabilization of vesicles and micelles. Work was performed on seven individual studies. Several different types of novel cleavable surfactants were synthesized, including systems with two chains and two head groups, and their aqueous micelles and vesicles were characterized by methods including dynamic laser light scattering (to give aggregate sizes) and differential scanning calorimetry (to give vesicle bilayer phase transition temperatures). A cleavable surfactant is stable under certain conditions but liable under other conditions with respect to cleavage to give nonsurfactant products and/or other surfactants with different properties. The destruction of several cleavable surfactant-derived aggregates ...
Understanding the physical characteristics of the local microenvironment in which Mycobacterium tuberculosis resides is an important goal that may allow the targeting of metabolic processes...Full Text Available
The susceptibility of 19 isolates of Haemophilus ducreyi from a recent chancroid outbreak and four reference strains was determined in vitro to 13 antimicrobial agents. The rabbit intradermal...Full Text Available
Acetylcholine-rich synaptic vesicles were isolated from myenteric plexus-longitudinal muscle strips derived from the guinea pig ileum by the method of Dowe, Kilbinger, and Whittaker [J. Neurochem. 35, 993-1003 (1980)] using either unstimulated preparations or preparations field-stimulated at 1 Hz for 10 min using pulses of 1 ms duration and 10 V . cm-1 intensity. The organ bath contained either tetradeuterated (d4) choline (50 microM) or [3H]acetate (2 muCi . ml-1); d4 acetylcholine was measured by gas chromatography-mass spectrometry. As with Torpedo electromotor cholinergic vesicle preparations made under similar conditions the distribution of newly synthesized (d4 or [3H]) acetylcholine in the zonal gradient from stimulated preparations was not identical with that of endogenous (d0, [1H]) acetylcholine, but corresponded to a subpopulation of denser vesicles (equivalent to the VP2 fraction from Torpedo) that had ...
An overview of current concepts on neuroinflammation and on the dialogue between neurons and non-neuronal cells in three important infections of the central nervous systems (rabies, cerebral malaria, and human African trypanosomiasis or sleeping sickness) is here presented. Large numbers of cases affected by these diseases are currently reported. In the context of an issue dedicated to Camillo Golgi, historical notes on seminal discoveries on these diseases are also presented. Neuroinflammation is currently closely associated with pathogenetic mechanisms of chronic neurodegenerative diseases. Neuroinflammatory signaling in brain infections is instead relatively neglected in the neuroscience community, despite the fact that the above infections provide paradigmatic examples of alterations o...
A case report of falsely elevated serum hormone values measured by radio-immunoassay (RIA) is described. The radio-immunoassays concerned have a first antibody raised in rabbits and mostly a separation technique based on a second antibody-solid phase system. The presence of heterophyle (anti-rabbit) antibodies in patients' serum is proved.
Eight rabbits and 24 guinea-pigs were exposed to a silica dust cloud of about 40,000 pp./ml. (0·3-3·0 μ) and killed at four-weekly intervals up to 30 weeks. The guinea-pigs showed...Full Text Available
Autophagy is a ubiquitous catabolic process that ensures organism’s well-being by sequestering a wide array of undesired intracellular constituents into double-membrane vesicles termed...Full Text Available
A method for the study of conjugated polyelectrolyte (CPE) photophysics in solution at the single-molecule level is described. Extended observation times of single polymer molecules are enabled by the...Full Text Available
New visual pigments were formed with 4-hydroxy retinals in isolated vertebrate rod photoreceptors by exposing bleached rods from the tiger salamander, Ambystoma tigrinum, to lipid vesicles containing...Full Text Available
X-ray diffuse scattering was measured from oriented stacks and unilamellar vesicles of dioleoylphosphatidylcholine lipid bilayers to obtain the temperature dependence of the structure and of the material...Full Text Available
Atg4 cysteine proteases (autophagins) play crucial roles in autophagy by proteolytic activation of Atg8 paralogs for targeting to autophagic vesicles by lipid conjugation, as well as in subsequent deconjugation...Full Text Available
The H+-translocating inorganic pyrophosphatase (H+-PPase) associated with vesicles of the vacuolar membrane (tonoplast) isolated from beet (Beta vulgaris L.) is...Full Text Available
Two methods were used to label pig kidney microvillar membrane proteins from the luminal and cytoplasmic surfaces of closed membrane vesicles. The first method was lactoperoxidase-catalysed radioiodination. Lactoperoxidase and glucose oxidase were positioned inside or outside the vesicles, iodination being initiated by adding glucose and "1"2"5I. After electrophoresis of the proteins, asymmetric labelling patterns on radioautographs were observed. However the major disadvantage of this method was the high degree of intramembrane labelling of the fatty acid chains of membrane lipids. The second method overcame this disadvantage. A new hydophilic photoreagent, 3,5-di("1"2"5I)iodo-4-azidobenzenesulphonate, was transported by a Na"+-dependent system into microvillar vesicles, thus permitting labelling from either side of the membrane when the vesicles were photolysed. The activity of several microvillar ...
A study was undertaken of the effect of monovalent cations (Li/sup +/, Na/sup +/, K/sup +/) on the dielectric constant (epsilon) of the water-lipid interface of unilamellar phosphatidylcholine (PC) vesicles, i.e., the ester carbonyl oxygen region of the PC molecules or the neighborhood of the oxygen atoms of the phosphorylcholine moiety. epsilon was determined by reacting the free radical 1,1-diphenyl-2-picrylhydrazyl with ..cap alpha..-tocopherol incorporated in the lipid vesicles. The results are consistent with a decrease of epsilon (LiCl: 35.5 to 29.5; NaCl: 34 to 29; KCl: 33 to 29) as the concentration of the salts in the solvent media increases from 0.025 to 0.5 M. These effects can be rationalized in terms of dielectric saturation at the water-lipid interface brought about by ion-induced local electric fields. In the unilamellar PC vesicles the effect of the ions on epsilon follows the sequence K/sup +/ > Na/sup ...
Exosomes are small membrane vesicles secreted from various types of cells. Tumor-derived exosomes contain MHC class I molecules and tumor-specific antigens, receiving attention as a potential cancer...Full Text Available
The delivery of proteins and organelles to the vacuole by autophagy involves membrane rearrangements that result in the formation of large vesicles called autophagosomes. The mechanism underlying autophagosome...Full Text Available
The curvature elastic modulus (bending stiffness) of stearoyloleoyl phosphatidylcholine (SOPC) bilayer membrane is determined from membrane tether formation experiments. R. E. Waugh and R. M. Hochmuth...Full Text Available
PurposeWe objectively quantified the gain in urethral diameter and the effect of stenting after tubularized incised plate urethroplasty in a rabbit hypospadias model. Materials and MethodsWe created a hypospadias model in 12 New Zealand white male rabbits by excising the ventral urethra. A 3 cm tattoo line was made longitudinally in the dorsal urethral plate midline. Two weeks later a 2 cm relaxing incision was made in the middle part of the tattooed line. The stretched incision width between the tattooed edges was measured, followed by urethral plate tubularization. Six rabbits were stented and 6 were nonstented. Two weeks later the animals were sacrificed and the distance separating the tattoo was measured at the midpoint of the tattooed line. Transverse sections at this point were exami...
In a series of experiments carried out on young and adult rabbits the effect of isocaloric low protein diets containing 4% or 8% protein compared with a diet containing 21% protein on Obeliscoides cuniculi infection was studied. The pathogenesis, resistance and local immunity were assessed after single infections with 10,000 larvae or reinfection with 5000 larvae. Live weight gain was reduced in young and adult rabbits fed the low protein diets, but the establishment of parasites was not substantially influenced by protein deprivation. However, development of worms in the histotrophic phase and parasite fecundity were impaired in association with the low protein diet. Moreover, mild anaemia as well as changes in the mucosal immune response as a result of infection were related to the level of dietary protein. (author). 30 refs, 6 figs, 5 tabs.
Experiments have been reported previously in which lead-212 ions with lead carrier were instilled into the lungs of anaesthetised rabbits. Experiments were also reported in which bismuth-207 chloride was instilled into rabbit lung. To check that the chloride compound was cleared in the same way as ions instilled in isolation, lead-203 chloride has been instilled in three rabbits. Measurements of absorption of ionic lead have been repeated using the radon-222 daughter, lead-214. Rates of clearance from the lung of "2"0"3Pb as the chloride and "2"1"2Pb ions are compared, and retention functions are derived. Clearance from the lung of "2"1"4Pb ions is also shown, and retention functions are derived. (U.K.).
A comparative and quantitative study of the tissue distribution of brain-thymus shared antigens was carried out using rabbit antisera to rat, dog and human brain homogenates, assayed on rat, dog and...Full Text Available
1. Seven unique carboxymethylcysteine-containing peptides have been isolated from tryptic digests of rabbit muscle aldolase carboxymethylated with iodo[2-14C]acetic acid in 8m-urea....Full Text Available
1. A novel phthalazine analogue taloximine (1-hydroxyimino-4(2-dimethyl-aminoethoxy)-1,2-dihydrophthalazine monohydrochloride monohydrate) stimulated respiration in conscious rabbits at doses of 7...Full Text Available
The pharmacological effects of a series of fatty acid-choline esters have been studied on the isolated rabbit heart, the isolated guinea-pig ileum and the rat stomach. The effect changed with increasing...Full Text Available
A regulatory locus on the Staphylococcus aureus chromosome, designated sar, is involved in the expression of cell wall proteins, some of which are potentially important in the pathogenesis of endocarditis....Full Text Available
Hyaluronan (HA) is important for joint cavitation, lubrication, volume regulation and synovial fluid drainage but little is known about the regulation of joint HA synthesis/secretion in vivo....Full Text Available
... Weiss, S., D. Kobiler, H. Levy, H. Marcus, A. Pass, N. Rothschild, and Z ... of Bacillus anthracis spores conferred by a protective antigen-based vaccine in rabbits ...
Naturally occurring, double-stranded RNA (ds-RNA)) was immunogenic when injected into mice, rats, guinea-pigs, rabbits, dogs and baboons. The response to native material administered intravenously (i.v.)...Full Text Available
An antiserum was raised in a rabbit against highly purified human liver dihydropteridine reductase (EC 1.6.99.7). Dihydropteridine reductase from human liver, in human cultured fibroblasts and in continuous...Full Text Available
Antibodies of the IgM type present in rabbit anti-epiglycanin antiserum were purified by (NH4)2SO4 precipitation and by ion-exchange, affinity and gel-filtration chromatography. After papain treatment...Full Text Available
ABSTRACTObjectivesTo test the feasibility of human immune globulin (IG, Gamimune N, 10%) as a new treatment for endophthalmitis, the ocular tolerance, distribution,...Full Text Available
Orientation and movement relies on both visual and vestibular information mapped in separate coordinate systems. Here, we examine how coordinate systems interact to guide eye movements of rabbits....Full Text Available
... The costal plain still supports Jarrah (Eucalyptus marginata ) forest, dense woodlands, coastal heath and diverse swamplands. Many species of plants are endemic. 20 species of mammals and 12 species of introduced mammals including with feral pigs, rabbits and foxes of particular concern....
1. Gamma globulin metabolism and distribution were studied employing rabbit gamma globulin (RGG)I(131) 24 times in 13 control rabbits. Similar studies were performed before and during the ananmestic response in 4 rabbits previously sensitized with a polyvalent pneumococcal vaccine. 2. During the ananmestic response, gamma-globulin levels increased from 1.0 to 6.0 gm/100 ml, and the gamma-globulin pool increased from 0.7 to 4.7 gm/kg. There was no change in the intravascular-extravascular partition of gamma globulin. 3. Gamma globulin degradation increased from 0.06 to 0.33 gm/kg/day during the 28 days of the immunization period while gamma globulin synthesis increased even further to average 0.47 gm/kg/day. Following the attainment of elevated gamma globulin levels the fractional rate of RGG-I(131) turnover increased from 8.0 to 12.5 per cent/day. 4. No differences were noted in the metabolism of homologous or autologous ...
Assessment of subtle changes in the primary macromolecular components of cartilage, proteoglycan (PG) and collagen, is critical for the diagnosis of early stages of osteoarthritis (OA), but...Full Text Available
Drugs used for the inhibition of DNA viruses, such as iododeoxyuridine, adenine arabinoside, or trifluorothymidine, are not biochemically selective in their action and also interfere with normal cellular...Full Text Available
ObjectivesWe examined the expression of Rho Kinase (ROK) isoforms in young and old rabbit’s detrusor smooth muscles (SM) during the progression of short term...Full Text Available
In vivo, mammalian lenses have the capacity to effect fully reversible changes in shape, and possibly volume, during the accommodation process. Isolated lenses also change shape...Full Text Available
Reaggregated cells from 6- to 8-day-old mouse cerebella have been used to raise antibodies in rabbits. The interaction of these antibodies with cerebellar cell surface components was assessed by cytotoxicity...Full Text Available
Tooth enamel biomineralization is mediated by enamel proteins synthesized by ameloblast cells. Two classes of proteins have been described: enamelins and amelogenins. In lower vertebrates the absence...Full Text Available
Polyclonal antibodies were raised in rabbits by immunization with free peptides corresponding to positions 197-222 of the human beta 1-adrenergic receptor (beta 1 peptide) and the corresponding sequence...Full Text Available
The placental transfer of a single intravenous injection of 14C-hexoprenaline was studied in eight pregnant New Zealand white rabbits. Maternal and fetal blood was sampled intermittently for 60 minutes after the injection. An initial rapid decrease in the levels of 14C-hexoprenaline in maternal blood was followed by a second slower phase, whereas fetal levels remained insignificant. The conclusion, therefore, is that the rapid improvement in fetal heart rate after the administration of a single maternal intravenous injection of hexoprenaline in the treatment of fetal distress is due to the action on the uterus and/or on maternal cardiovascular function, and not to direct stimulation of the fetus.
The seminal work of Stern and his coauthors on integrated control has had a profound and long-lasting effect on the development of IPM programs in western orchard systems. Management systems based solely on pesticides have proven to be unstable, and the success of IPM systems in western orchards has been driven by conservation of natural enemies to control secondary pests, combined with pesticides and mating disruption to suppress the key lepidopteran pests. However, the legislatively mandated changes in pesticide use patterns prompted by the Food Quality Protection Act of 1996 have resulted in an increased instability of pest populations in orchards because of natural enemy destruction. The management system changes have made it necessary to focus efforts on enhancing biological control n...
A renewed interest in chemical fractionation of soil organic matter (SOM) originates from the premise that it enables to isolate labile SOM from SOM protected through mineral binding and recalcitrant SOM. Both selective removal of labile non-bound SOM through oxidation or hydrolysis as well as selective removal of minerals and attached SOM are often applied. Molecular-level SOM characterization by means of temperature resolved Pyrolysis-Field Ionization Mass Spectroscopy analysis (Py-FIMS) was used here as an approach to obtain insight into the fate of SOM upon wet chemical treatment with regard to composition and thermal stability. The applied sequential chemical treatment with 6% NaOCl and 10% HF yielded similar sizes in stable SOM fractions between sandy semi-native heathland and cultiv...
As evidence mounts that male genitalia can affect relative fertilisation success, the role that sexual selection has played in the rapid and divergent evolution of genitalia is becoming increasingly recognized. Unfortunately, the limited functional understanding of these complex structures and their interactions with the female reproductive tract often limit interpretation regarding their evolution. Here, we address this issue using the earwig Euborellia brunneri, where both the male intromittent organ and the female spermatheca are highly exaggerated in length yet structurally simple. In a double mating design, we use the sterile male technique to study how sperm precedence patterns are affected by male genital length, male age, and the size of the male sperm storage organ, the seminal ve...
Objective: To evaluate technical feasibility and acute and subacute radiotolerance of a self-expandable stent loaded with {sup 125}I seeds in the rabbit esophagus. Methods: A self-expandable stent designed for esophageal application was made of 0.16 mm nitinol wire and loaded with {sup 125}I seeds (CIAE-6711). Twenty-seven stents with three different radioactive dosages (n = 9 in each dosage group) were implanted in the esophagus of healthy rabbits, while nine stents alone were used as controls. The stents were perorally deployed into the esophagus under fluoroscopic guidance. Radiological follow-up included plain chest film, CT scan, and barium esophagography which were undertaken in all rabbits of each group at 2, 4, and 8 weeks, respectively, which were correlated to histopathological findings. The stented esophageal segments along with their adjacent tissues were harvested for histopathological examinations. Results: ...
The influence of membrane pH gradients on the transbilayer distribution of some common phospholipids has been investigated. The authors demonstrate that the transbilayer equilibrium of the acidic phospholipids egg phosphatidylglycerol (EPG) and egg phosphatidic acid (EPA) can be manipulated by membrane proton gradients, whereas phosphatidylethanolamine, a zwitterionic phospholipid, remains equally distributed between the inner and outer monolayers of large unilamellar vesicles (LUVs). Asymmetry of EPG is examined in detail and demonstrated by employing three independent techniques: ion-exchange chromatography, {sup 13}C NMR, and periodic acid oxidation of the (exterior) EPG headgroup. In the absence of a transmembrane pH gradient ({Delta}pH) EPG is equally distributed between the outer and inner monolayers of LuVs. When vesicles composed of either egg phosphatidylcholine (EPC) or DOPC together with 5 mol % EPG are prepared with a transmembrane ...
The goal of the present study is to elucidate the effect of lipid domain formation on activities of Naja naja atra and Bungarus multicinctus phospholipase A(2) (PLA(2)) enzymes. Sphingomyelin inhibited enzymatic activity and membrane-damaging activity of PLA(2) against egg yolk phosphatidylcholine (EYPC), while cholesterol and cholesterol sulfate abrogated the inhibitory effect of sphingomyelin. The ability of cholesterol and cholesterol sulfate to abolish the inhibitory effect of sphingomyelin was closely related to their capacity to induce domain formation in EYPC/sphingomyelin vesicles. Laurdan fluorescence measurement revealed that membrane packing of EYPC/sphingomyelin vesicles was differently affected by cholesterol and cholesterol sulfate. Unlike cholesterol, cholesterol sulfate was unable to promote domain formation in dipalmitoylphosphatidylcholine (DPPC) vesicles. Cholesterol increased but cholesterol sulfate ...
Asphalt emulsions were evaluated as a means to immobilize radiological contaminants deposited on urban surfaces after a dirty bomb attack. Contaminated surfaces would be sprayed with thin coatings of asphalt emulsion to encapsulate the radioactive particles until the site can be safely remediated. This research investigated applications of an asphalt emulsion (Topein C, Encapco Technologies, LLC, Napa, CA) to treat (zero-valent) iron, lead, and uranium powders on various building material surfaces. Initial studies found that some of the building materials (limestone, concrete, and metal) reacted with the emulsion producing gas bubbles, which formed 0.001 to 1 cm vesicles in the cured asphalt emulsion. These vesicles, however, did not expose the building material surface, and the reaction a...
The integral membrane protein synaptophysin is one of the most abundant polypeptide components of synaptic vesicles. It is not essential for neurotransmission despite its abundance but is believed to modulate the efficiency of the synaptic vesicle cycle. Detailed behavioral analyses were therefore performed on synaptophysin knockout mice to test whether synaptophysin affects higher brain functions. We find that these animals are more exploratory than their wild type counterparts examining novel objects more closely and intensely in an enriched open field arena. We also detect impairments in learning and memory, most notably reduced object novelty recognition and reduced spatial learning. These deficits are unlikely caused by impaired vision, since all electroretinographic parameters measur...
Recent studies have shown Y chromosome microdeletions associated with male infertility. The factors responsible for Y chromosome microdeletions in spermatozoa remain unresolved. However, the environmental pollutants are known to damage DNA in differentiating and maturing germ cells in the male reproductive tract. Therefore, the aim of this study was to investigate the effects of seminal hexachlorocyclohexane (HCH) and its isomers, an environmental pollutant, in 50 fertile and 50 infertile males in relation to semen quality and the incidence of Y chromosome microdeletion in azoospermic factor (AZF) region. As compared to control, an increased HCH level and significantly decreased semen quality were observed in the infertile males. A positive significant association was found between sperm count with alpha-HCH and beta-HCH in the infertile males. A negative significant association was observed between sperm counts with gamma-HCH in asthenospermia patients and with ...
To assess the technical feasibility of a newly designed stent-like electrode in rabbits. A stent-like electrode was knitted from a single thread of nitinol wire and interconnected to a generator using similar wire. In order to gauge the extent of radiofrequency ablation (RFA), we measured the depth of the ablated area in cow liver using a combination of 180-sec time intervals and 20- watt power increments. For data processing, Cox regression analysis was used. RFA was also applied to the small intestine of rabbits using this stent-like electrode under six different sets of conditions: 10 watts for 1 min, 10 watts for 2 mins, 20 watts for 1 min, 20 watts for 2 mins, 30 watts for 1 min, and 30 watts for 2 mins. To determine the gross and microscopic findings, six animals were sacrificed immediately after the procedure and the results obtained under the different sets of conditions were correlated. Eight rabbits were monitored ...
The water soluble, photolabile nitrene precursor, azidonaphthalene-2,7-disulfonic acid (ANDS) was encapsulated in small unilamellar, isoelectric (egg PC) or negatively charged (egg PC + dihexadecylphosphate) liposomes. The individual and combined effects of heme-proteins and UV irradiation on the fluorescence of these vesicles under aerobic conditions were studied. Consistent with the catalytic action of heme-proteins on lipid peroxidation and peroxide decomposition, addition of cytochrome c (positively charged) or catalase (negatively charged) to the vesicles elicited immediate formation of a fluorescence band at 470 nm, characteristic of Schiff bases that form from aldehyde byproducts of decomposing hydroperoxides. Ultraviolet irradiation of liposomes caused no significant changes in the fluorescence spectrum, in spite of the radiolysis of ANDS inside the vesicles with consequent formation of nitrene radicals. When ...
The presynaptic Ca2+ signal, which triggers vesicle release, disperses to a broadly distributed residual [Ca2+] ([Ca2+]res) that plays an important...Full Text Available
Arf GTPases control vesicle formation from different intracellular membranes and are regulated by Arf guanine nucleotide exchange factors (GEFs). Outside of their conserved catalytic domains, known...Full Text Available
Calcitonin gene-related peptide (CGRP) was localized by an immunocytochemical technique in the thyroid-parathyroid complexes of rat, guinea pig, rabbit, and in normal human thyroids and parathyroids. Human medullary carcinomas and parathyroid adenomas were also studied. In man and all animal species examined CGRP was present in the parafollicular cell, however, in guinea pigs only in small amounts. Except in rabbits, presence of CGRP was demonstrated in nerves of the thyroid and parathyroid capsule as well as in the nerve fibers of the capsular blood vessels. In the thyroid of guinea pigs CGRP was also noted in nerve fibers and in blood vessel walls between follicles. CGRP was also present in the parathyroid glands of rat and man, in nerve fibers localized between parathyroid cells. In rabbit the parafollicular cells between parathyroid cells also expressed CGRP immunoreactivity. No CGRP was noted in the parathyroids of the ...
Glycosaminoglycans synthesized by cultured rabbit smooth muscle cells were isolated after incorporation of (/sup 3/H)-glucosamine into glycosaminoglycans in the presence or absence of 10% fetal bovine serum. Glycosaminoglycans were quantitated by two-dimensional electrophoresis after proteolytic digestion of the cell layers and media. The results show that the presence of serum has no effect on the chondroitin sulfate, heparan sulfate and dermatan sulfate content of the cell layers. The incorporation of (/sup 3/H)-glucosamine into hyaluronic acid of the cell layers was three times higher in the presence of serum. In the medium , the quantity of hyaluronic was two times higher in the presence of serum while the other glycosaminoglycans remained unchanged. The incorporation of (/sup 3/H)-glucosamine into hyaluronic acid was unaffected by the presence of serum. Specific proteoglycans were isolated from medium after with (/sup 35/S)-sulfate and (/sup 3/H)-serine by ...
A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of and)2numberSPO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the TUPO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide and protein substrate specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from diabetic rabbits was phosphorylated in vitro by this multifunctional kinase at only 10% of the rate compared to synthase ...
Purpose: The lifelong persistence of foreign bodies within the arteries may contribute to restenosis. Thus,biodegradable devices might decrease recurrence rates. Methods: Eleven polyhydroxybutyrate biodegradable stents and 13 tantalum stents were implanted into the iliac arteries of New Zealand white rabbits for up to 30 weeks. After killing the animals,the specimens were harvested, fixed in formalin, processed in paraffin,and stained. Results: Polyhydroxybutyrate instigated intense inflammatory and proliferative reactions with an increase in collagen (2.4- to 8-fold vs native segments), thrombosis and in-stentlumen narrowing (375.5-606.6 mm vs 655.6 #+-# 268.8 mm in native segments). The elastic membranes were destroyed in all specimens. The tantalum stents increased the in-stent lumen progressively (769.7 #+-# 366.6 mm vs 1309.9 #+-# 695.3 mm),penetrated the external elastic membrane, and increased mural collagen content (6- to 8.6-fold vs native segments). ...
Abstract Purpose: We investigated the effects of various rinsing and healing protocols on corneal wound repair and inflammation following alkali burn in rabbits. Methods: We conducted in vitro, in vivo and ex vivo studies. First, different rinse solutions were tested in vitro after incubation of ocular cells with methanol or NaOH. Cell viability was then assessed using the neutral red test (cytofluorometry). Second, NaOH was applied to rabbit corneas and associations of rinse solutions (NaCl 0.9% or controlled ionization marine solutions) with N-acetylcysteine or vegetable oils (from Calophyllum inophyllum and Aleurites moluccana) were tested in vivo. The regeneration of the corneal epithelium and the infiltration of inflammatory cells were evaluated using in vivo confocal microscopy and e...
The purpose of this study was to investigate whether hydroxyapatite (HAp) coating could induce polyethylene terephthalate (PET) artificial ligament graft osseointegration in the bone tunnel. Twenty-four New Zealand white rabbits underwent artificial ligament graft transplantation in bilateral proximal tibia tunnels. One limb was implanted with HAp-coated PET graft, and the contralateral limb was implanted with non-HAp-coated PET graft as control. The rabbits were randomly sacrificed at four and eight?weeks after surgery. The loads to failure of the experimental group at eight?weeks were significantly higher than those of the control group (p?=?0.0057). Histologically, application of HAp coating induced new bone formation between graft and bone at eight?weeks compared with the controls. Rea...
Purpose: To evaluate the therapeutic effect of heated (60 deg. C) lipiodol via hepatic artery administration in a rabbit model of VX2 liver cancer. Materials and methods: Thirty male New Zealand white rabbits were randomly divided into three groups with 10 rabbits assigned to each group. VX2 carcinoma cells were surgically implanted into the left hepatic lobe. The tumors were allowed to grow for 2 weeks, and studies were performed until the diameter of the tumors detected by ultrasonograph reached 2-3 cm. Under anesthesia, trans-catheter hepatic arterial embolization was performed and doxorubicin-lipiodol (37 deg. C) (1 mL), lipiodol (60 deg. C) (1 mL) or control (physiological saline (37 deg. C) (1 mL)) solution was injected into the hepatic arteries of animals in the three groups. One week later, the volume of the tumor was measured by ultrasonograph again. The serum of all rabbits was collected ...
Aim: This study was performed: 1) to assess the relationship between blood flow velocity in the internal carotid artery (CBFICA) and pial artery pulsation (cc-TQ) and 2) to evaluate flow-induced changes in pial artery compliance. Methods: Experiments were performed on 10 crossbred male rabbits. Heart rate (HR), blood pressure (BP), left ventricle ejection fraction (LVEF), CBFICA, the systolic-diastolic blood volume fraction in the brain circulation (CBFSDF) and cc-TQ were recorded after glucagon and acetazolamide administration. cc-TQ was measured with near-infrared transillumination back scattering sounding (NIR-T/BSS), LVEF and CBFSDF with gated scintigraphy and BP and CBFICA with electromagnetic pressure and flow transducers, respectively. Doses of drugs were chosen to exert a haemodyna...
Uteroglobin (UG) gene encodes a cytokine-like, multifunctional, antiinflammatory protein, with potent phospholipase A2-inhibitory activity. It has been suggested that during implantation this protein...Full Text Available
... The dawn of Bio-Mecha and a Biomechanical Revolution May 31 2011: Hi Julian, Yes, but i dont think we would ... The dawn of Bio-Mecha and a Biomechanical Revolution May 31 2011: A BBC Documentary: All Watched Over by Machines of Loving ... The dawn of Bio-Mecha and a Biomechanical Revolution May 17 2011: Ed Boyden: A light switch for neurons is an amazing ... The dawn of Bio-Mecha and a Biomechanical Revolution May 16 2011: Thank you Meher Like Spring Rabbit, its a great contribution....
The use of tick vaccines in mammalian hosts has been shown to be the most promising alternative tick control method to current use of acaricides, which suffers from a number of limitations. However,...Full Text Available
BackgroundHerpes simplex virus type-1 (HSV-1) infections can cause a number of diseases ranging from simple cold sores to dangerous keratitis and lethal encephalitis. The interaction...Full Text Available
Correlates between immunological parameters and protection against Bacillus anthracis infection in animals vaccinated with protective antigen (PA)-based vaccines could provide surrogate markers to evaluate the putative protective efficiency of immunization in humans. In previous studies we demonstrated that neutralizing antibody levels serve as correlates for protection in guinea pigs (S. Reuveny et al., Infect. Immun. 69:2888-2893, 2001; H. Marcus et al., Infect. Immun. 72:3471-3477, 2004). In this study we evaluated similar correlates for protection by active and passive immunization of New Zealand White rabbits. Full immunization and partial immunization were achieved by single and multiple injections of standard and diluted doses of a PA-based vaccine. Passive immunization was carried out by injection of immune sera from rabbits vaccinated with PA-based vaccine prior to challenge with B. anthracis spores. Immunized ...
An analogue of hGH isolated from plerocercoids of the tapeworm Spirometra mansonoides displaces ["1"2"5I]hGH from its receptors in rabbit, rat, and hamster liver membranes. Biologically, plerocercoid growth factor (PGF) is more similar to hGH than to other mammalian GH's but has not been shown to bond human cells. Receptors specific for hGH have been described on cultured human lymphocytes (IM-9). In this study, the authors compared the binding of PGF and hGH in IM-9 cells and in rabbit hepatic membranes. IM-9 lymphocytes (12 x 10"6 cells/tube) were incubated with ["1"2"5I]hGH and increasing concentrations of hGH (ng/ml) or PGF (serial dilutions) for 90 min at 30"0 C. Specific binding (B_0 - NSB) was determined for each dose of hGH or PGF and the binding curves were analyzed by logit-log regression. The results show that PGF displaced ["1"2"5I]hGH from human cells in a dose dependent manner (r = 0.98). Based on the IM-9 assay, 1 ml of the PGF ...
1. Amino acid sequences covering the region between residues 173 and 248 [adopting the numbering system proposed by Lai, Nakai & Chang (1974) Science 183, 1204-1206] were derived for trout (Salmo...Full Text Available
The rates of spontaneous interbilayer and transbilayer exchange of ["3H]dimyristoylphosphatidylcholine (["3H]DMPC) were examined in the DMPC and DMPC/dimyristoylphosphatidylethanolamine (DMPE) large unilamellar vesicles in the liquid-crystalline-, gel-, and mixed-phase states. DMPC desorption rates from either gel or liquid-crystalline phases containing DMPE are very similar to the corresponding rates from pure DMPC gel or liquid-crystalline phases. The authors proposed that the DMPC/DSPC behavior results from packing defects in gel phases composed of both DMPC and DSPC molecules because of the four-carbon difference in the acyl chain lengths of the two species. The present results strongly support this hypothesis because no such anomalous behavior is observed in DMPC/DMPE, which is similar to DMPC/DSPC in phase behavior but does not have the chain length difference. These results are not consistent with lipid headgroup dehydration as the major energetic barrier to ...
The temporal development of the effect of THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) on the ultrastructure composition and GABA receptor expression in cerebellar granule cells was investigated by quantitative electron microscopy (morphometric analysis) and GABA binding assays. It was found that the cytoplasmic density of smooth endoplasmic reticulum was decreased, while the cytoplasmic density of rough endoplasmic reticulum, Golgi apparatus, vesicles and coated vesicles was greatly enhanced after exposure of the cells to THIP (150 microM) for only 1 hr. In cerebellar granule cells exposed to THIP (150 microM) for 3 hr low affinity GABA receptors were induced. These findings show that the effect of THIP on the ultrastructure composition and GABA receptor expression in cultured cerebellar granule cells may be interrelated and moreover it is likely that the turn-over of GABA receptors is extremely fast.
This project has received DOE support since July 1, 1978. During this period, 40 papers have been published dealing predominantly with chlorophyll-photosensitized electron transfer reactions in a variety of media (solutions, polymer films, lipid bilayer membranes). The overall theme of this work has been to develop mechanistic strategies f or photochemical energy storage via chlorophyll, using the green plant photosynthetic system as a paradigm for designing in vitro systems. Microheterogeneous lipid vesicle suspensions allow ready application of time-resolved optical spectroscopy to follow the course of light-induced electron transfer processes. Both the yields and the lifetimes of electron transfer products were markedly improved in the vesicle systems. In subsequent studies, this compartmentalization was favorably manipulated by controlling the electrical charge on the membrane surface, by controlling the solubility properties of the ...
Synaptobrevin is a synaptic vesicle protein that has an essential role in exocytosis and forms the SNARE complex with syntaxin and SNAP-25. We have analyzed the structure of isolated synaptobrevin and its binary interaction with syntaxin using NMR spectroscopy. Our results demonstrate that isolated synaptobrevin is largely unfolded in solution. The entire SNARE motif of synaptobrevin is capable of interacting with the isolated C-terminal SNARE motif of syntaxin but only a few residues bind to the full-length cytoplasmic region of syntaxin. This result suggests an interaction between the N- and C-terminal regions of syntaxin that competes with core complex assembly.
The murine 3T3-L1 fibroblast under appropriate incubation conditions differentiates into an adipocyte phenotype. This 3T3-L1 adipocyte exhibits many of the morphologic, biochemical, and insulin-responsive features of the normal rodent adipocyte. Using quantitative electron microscopic (EM) autoradiography we find that, when /sup 125/I-insulin is incubated with 3T3-L1 adipocytes, the ligand at early times of incubation localizes to the plasma membrane of the cell preferentially to microvilli and coated pits. When the incubation is continued at 37 degrees C, /sup 125/I-insulin is internalized by the cells and preferential binding to the villous surface is lost. With the internalization of the ligand, two intracellular structures become labeled, as determined by the method of hypothetical grain analysis. These include large clear, presumably endocytotic, vesicles and multivesicular bodies. Over the first hour of incubation the labeling of these structures increases in ...
The effects of purified Helminthosporium maydis T (HmT) toxin on active Ca/sup 2 +/ transport into isolated mitochondria and microsomal vesicles were compared for a susceptible (T) and a resistant (N) strain of corn (Zea mays). ATP, malate, NADH, or succinate could drive /sup 45/Ca/sup 2 +/ transport into mitochondria of corn roots. Ca/sup 2 +/ uptake was dependent on the proton electrochemical gradient generated by the redox substrates or the reversible ATP synthetase, as oligomycin inhibited ATP-driven CA/sup 2 +/ uptake while KCN inhibited transport driven by the redox substrates. Purified native HmT toxin completely inhibited Ca/sup 2 +/ transport into T mitochondria at 5 to 10 nanograms per milliliter while transport into N mitochondria was decreased slightly by 100 nanograms per milliliter toxin. Malate-driven Ca/sup 2 +/ transport in T mitochondria was frequently more inhibited by 5 nanograms per milliliter toxin than succinate or ATP-driven Ca/sup 2 +/ ...
The authors used radioligand binding and studies of cell contraction to characterize muscarinic receptors on dispersed smooth muscle cells from rabbit proximal and distal colon. Cells obtained after serial incubations in collagenase were used to measure binding of tritiated quinuclidinyl benzilate (["3H]QNB). At 37 degree C, specific ["3H]QNB binding was saturable and linearly related to cell number. Nonlinear regression analysis was used to determine the affinity of ["3H]QNB for its receptor. The IC_5_0 for the muscarinic agonists bethanechol and oxotremorine were 80 and 0.57 #mu#M, respectively. Hill coefficients were 0.67 for both, suggesting more complex interaction involving receptors of different affinities. In studies of cell contraction, bethanechol stimulated a dose-dependent decrease in cell length with half the maximal contraction occurring at 100 pM. These results suggest that (1) contraction is mediated by binding of bethanechol to M_2-muscarinic ...
Hyaluronan is an important constituent of the extracellular matrix in skin, and recent studies suggest that there is a pool of easily removable (free) hyaluronan drained by lymph. The removal rate of free hyaluronan in skin was measured from the elimination of ({sup 3}H)hyaluronan, injected subcutaneously in 13 rabbits. The removal of radioactivity was determined from appearance of {sup 3}H in plasma. During the first 24 h after injection, 10-87% of the tracer entered blood, less in injectates with high concentrations of hyaluronan. The removal was monoexponential with a half-life of 0.5-1 day when concentration of hyaluronan was 5 mg/ml or less. When hyaluronan concentration was 10 mg/ml or higher, the removal was slow for about 24 h and then became similar to that in experiments with low hyaluronan concentration. Free hyaluronan at physiological concentrations is thus turned over with the same rate as serum albumin, supporting the concept that hyaluronan is ...
Rat lung and liver microsomes did not undergo lipid peroxidation in the absence of iron when incubated with NADPH and concentrations of paraquat ranging from 10/sup -7/ to 10/sup -2/ M. Paraquat also did not stimulate rat liver and lung microsomal peroxidation induced by added iron and NADPH, and was inhibitory at concentrations above 10 ..mu..M. Similarly, no stimulation of peroxidation was produced by paraquat in rabbit or human lung microsomes; however, under similar conditions, paraquat enhanced NADPH/iron-dependent peroxidation in mouse lung and liver microsomes obtained from rats sacrificed at 12, 18, and 24 hr following a lethal dose of paraquat (50 mg/kg, ip), there was no loss of vitamin E or increase in susceptibility to in vitro peroxidation which would be expected if lipid peroxidation had occurred in vivo although extensive lung damage developed during this time period. These results indicate that paraquat does not cause pulmonary toxicity by ...
A new /sup 99m/Tc-phosphine-isocyanide complex with the general structure (/sup 99m/Tc (DEPE)/sub 2/(CNR)/sub 2/)/sup +/ has been synthesised and tested in animals and one human. In three animal species (rat, rabbit, dog), the complex is an efficient myocardial imaging agent, while in humans it remains in the blood pool. The complex is 100% protein bound in animals and humans, but whereas in humans it is attached to a 51.5 kdalton protein (probably prealbumin), in rabbits it appears to be bound to a larger macromolecule (M.W.>100 kdalton). The efficiency of the complex for blood pool labelling was tested in a human volunteer and compared with the standard in vivo red cell labelling technique with stannous pyrophosphate. A satisfactory radionuclide angiogram could be performed with less than 370 MBq of the complex. The count rate for the complex (cps/MBq) was 15% higher than that obtained with the labelled red cells and the absence of splenic ...
The placental transfer of a single intravenous injection of 14C-hexoprenaline was studied in eight pregnant New Zealand white rabbits. Maternal and fetal blood was sampled intermittently for 60 minutes after the injection. An initial rapid decrease in the levels of 14C-hexoprenaline in maternal blood was followed by a second slower phase, whereas fetal levels remained insignificant. The conclusion, therefore, is that the rapid improvement in fetal heart rate after the administration of a single maternal intravenous injection of hexoprenaline in the treatment of fetal distress is due to the action on the uterus and/or on maternal cardiovascular function, and not to direct stimulation of the fetus.
Central penetrating excisional wounds were made in the corneas of 12 rabbits and 10 trout. The scar tissue and the surrounding cornea were compared using a new method for assessing inorganic elements in the cornea: the scanning electron microscopy and energy dispersive analysis of x-ray (EDAX). Semiquantitative determination of inorganic elements within the range of atomic numbers 9-93 in the periodic system was performed, comparing the relative concentration of those elements in the scar tissue to the surrounding cornea. Results showed that calcium was the only element higher in the healing wound than in the surrounding cornea.
The plerocercoid stage of the tapeworm Spirometra mansonoides produces a functional analog of human growth hormone (hGH). Among the similarities between plerocercoid growth factor (PGF) and hGH is competition for the same receptors on rabbit liver membranes. To take advantage of this characteristic in a purification scheme for PGF, rabbit liver microsomes were solubilized in Triton X-100 and the hGH receptors were purified over an hGH affinity column. The purified receptors from six rabbit livers were coupled to Affi-Gel-10 to create a receptor affinity column which was used to purify PGF. Chromatography of crude PGF over the receptor column resulted in a 1044 fold increase in specific activity. SDS-PAGE in the presence of 2-mercaptoethanol showed that the affinity-purified PGF contained three protein bands with apparent Mrs of 27.5 K, 22 K, and 16.7 K. Injections of the partially-purified PGF into hypophysectomized rats ...
We irradiated proton beams on the ears of rabbits and the Harding-Passey mouse melanoma and observed their morphological change. We used 52 MeV proton beams from the INS-FM cyclotron. We adjusted the energy of the proton beams to be at the plateau part of the Bragg curve, at the half-way point of the Bragg peak, and at the Bragg peak. The amount of radiation was 5000rad in each case. The Harding-Passey mouse melanoma was transplanted into the subcutan of a three week old mouse. In this experiment, we used tumors, the diameter of which grew up to 1.5-2cm in 2-3 weeks after the transplantation. Using the jroscope, we observed both lightly and severely damaged cells. Using proton irradiation with the Bragg peak located at depth of 1mm in the rabbit's ears, we studied the change in the tissue. Irradiated epidermis fell off and was eroded because of radiation damage, but the rear surface of the rabbit's ears ...
A comparison of the concentration-response effects of inhaled ozone (O/sub 3/) and phosgene (COCl/sub 2/) in different species of laboratory animals was made in order to better understand the influence of the choice of species in inhalation toxicity studies. The effect of 4-h exposures to ozone at concentrations of 0.2, 0.5, 1.0, and 2.0 ppm, and to COCl/sub 2/ and 0.1, 0.2, 0.5, and 1.0 ppm was determined in rabbits, guinea pigs, rats, hamsters, and mice. Lavage fluid protein (LFP) accumulation 18-20 h after exposure was used as the indicator of O3- and COCl/sub 2/-induced pulmonary edema. All species had similar basal levels of LFP (250-350 mg/ml) when a volume of saline that approximated the total lung capacity was used to lavage the collapsed lungs. Ozone effects were most marked in guinea pigs, which showed significant effects at 0.2 ppm and above. Mice, hamsters, and rats showed effects at 1.0 ppm O3 and above, while rabbits responded ...
OBJECTIVE To present our initial experience of thulium laser resection via a flexible cystoscope for recurrent non-muscle-invasive bladder cancer (ThuRBT), as transurethral resection for bladder tumour (TURBT) is regarded as the reference standard for treating this disease, but alternative laser resection or ablation is suitable especially for recurrent tumours. PATIENTS AND METHODS From January 2005 to October 2005, 32 patients with early recurrent bladder tumour (recurrent within a year after TURBT) were treated with ThuRBT via a flexible cystoscope. The follow-up included urine analysis, ultrasonography and cystoscopy every 3 months. RESULTS All patients were treated successfully with ThuRBT in one session, with no bladder haemorrhage, obturator nerve reflex or vesicle perforation. Rand...
A new renicolid cercaria, Cercaria kuwaitae X sp. n., from the prosobranch gastropod Cerithium scabridum from Kuwait Bay is described. The new cercaria is nonstyleted gymnocephalous with voluminous Y-shaped excretory vesicle, flame cell formula 2[(3 + 3 + 3) + (3 + 3 + 3)] = 36, and without caudal fins. Surface topography of the new cercaria is studied by scanning electron microscopy. Domed papillae are abundant on the anterior and lateral aspects of the body, ciliated papillae are sparse dorsally. The ventral sucker bears nine domed papillae. Domed papillae form two circles around the oral sucker with three circles of spines in between. This is the first renicolid cercaria to be recorded in a gastropod from the Arabian Gulf region. PMID:9789783
Spatial Fluorescence Cross Correlation Spectroscopy is a rarely investigated version of Fluorescence Correlation Spectroscopy, in which the fluorescence signals from differ-ent observation volumes are cross-correlated. In the reported experiments, two observation volumes, typically shifted by a few $\\mu$m, are produced, with a Spatial Light Modulator and two adjustable pinholes. We illustrated the feasibility and potentiality of this technique by: i) measuring molecular flows, in the range 0.2 - 1.5 $\\mu$m/ms, of solutions seeded with fluorescent nanobeads or rhodamine molecules (simulating active transport phenomenons); ii) investigating the perme-ability of phospholipidic membrane of Giant Unilamellar Vesicles versus hydrophilic or hydrophobic molecules (in that case the laser spots were set on both sides of the mem-brane). Theoretical descriptions are proposed together with a discussion about FCS based, alternative methods.
An amphiphilic azobenzene diblock copolymer {2-[4-(4-methoxy phenyl azo)phenoxy]hexyl acrylate co-acrylic acid} was synthesized via reversible addition-fragmentation chain transfer polymerization in a hydrothermal reactor. The products were characterized by 1H-NMR, DSC, GPC and UV-Vis spectroscopy. Different self-assembly behaviors of this amphiphilic diblock copolymer in different organic solvent with different water content were investigated. In THF/H2O solution, the diblock copolymer self-assembled into spheres and the spheres would then change to vesicles with smaller size by increasing the volume of H2O. However, in DMSO/H2O solution, the sizes and morphologies of the self-assembled spheres would not change significantly with an enhanced amount of H2O. Photochromic behaviors of the am...
Abstract Septins are highly conserved cytoskeletal GTP-binding proteins implicated in numerous cellular processes from apoptosis to vesicle trafficking. Septins have been associated with leukemia and solid tumor malignancies, including breast, ovarian, and prostate. We previously reported that high SEPT9_i1 expression in human mammary epithelial cell lines (HMECs) led to malignant cellular phenotypes such as increased cell proliferation, invasiveness, motility, and genomic instability. Our goal here was to better understand how SEPT9_i1 expression might contribute to genomic instability and malignant progression. First, we confirmed that even transient expression of SEPT9_i1 was sufficient to increase aneuploidy in HMECs. We then analyzed SEPT9_i1 by immunoprecipitation and immunofluoresce...
The pore structure of volcanic clasts is examined using measurements of porosity, permeability, and electrical properties. Permeability varies by several orders of magnitude among volcanic clasts and does not depend solely upon porosity. Electrical property measurements of saturated volcanic samples illustrate the influence of pathway tortuosity and pore shape on permeability. For equivalent eruption conditions, silicic samples show higher tortuosities, smaller vesicle sizes, and lower permeabilities than mafic samples. These differences are largely due to variations in vesiculation and crystallization history. Differences between explosive and effusive samples reflect the relative ability of bubbles to form and maintain connected pathways during bubble expansion and collapse. Isotropic sa...
Chitosan (a?-(1-4)-amino-2-deoxy-b?-D-glucan) is a deacetylated form of chitin, a polysaccharide from crustacean shells. Its unique characteristics, such as positive charge, biodegradability, biocompatibility, nontoxicity, and rigid structure, make this macromolecule ideal for an oral vaccine delivery system. We prepared reverse-phase evaporation vesicles (REVs) sandwiched by chitosan (Chi) and polyvinylic alcohol (PVA). However, in this method, there are still some problems to be circumvented related to protein stabilization. During the inverted micelle phase of protein nanoencapsulation, hydrophobic interfaces are expanded, leading to interfacial adsorption, followed by protein unfolding and aggregation. Here, spectroscopic and immunological techniques were used to ascertain the effects ...
To examine the hormonal regulation of the ATP-dependent Ca{sup 2+} pump in the kidneys, the ATP-dependent Ca{sup 2+} uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ({sup 45}Ca{sup 2+}) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V{sub max} of the Ca{sup 2+} pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca{sup 2+} pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca{sup 2+} pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca{sup 2+} pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).
To examine the hormonal regulation of the ATP-dependent Ca"2"+ pump in the kidneys, the ATP-dependent Ca"2"+ uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ("4"5Ca"2"+) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V_m_a_x of the Ca"2"+ pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca"2"+ pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca"2"+ pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca"2"+ pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).
The synapsin III gene, SYN3, which belongs to the family of synaptic vesicle-associated proteins, has been implicated in the modulation of neurotransmitter release and in synaptogenesis, suggesting a potential role in several neuropsychiatric diseases. The human SYN3 gene is located on chromosome 22q12-13, a candidate region implicated in previous linkage studies of schizophrenia. However, association studies of SYN3 and schizophrenia have produced inconsistent results. In this study, four SYN3 SNPs (rs133945 (-631 C>G), rs133946 (-196 G>A), rs9862 and rs1056484) were tested in three sets of totally 3759 samples that comprise 655 affected subjects and 626 controls in the Irish Case-Control Study of Schizophrenia (ICCSS), 1350 samples incorporating 273 pedigrees in the Irish Study of High D...
Gamma-aminobutyric acid type A receptor-associated protein (GABARAP) belongs to a family of small ubiquitin-like adaptor proteins implicated in intracellular vesicle trafficking and autophagy. We have used diffusion-ordered nuclear magnetic resonance spectroscopy to study the temperature and concentration dependence of the diffusion properties of GABARAP. Our data suggest the presence of distinct conformational states and provide support for self-association of GABARAP molecules. Assuming a monomer-dimer equilibrium, a temperature-dependent dissociation constant could be derived. Based on a temperature series of {sup 1}H{sup 15}N heteronuclear single quantum coherence nuclear magnetic resonance spectra, we propose residues potentially involved in GABARAP self-interaction. The possible biological significance of these observations is discussed with respect to alternative scenarios of oligomerization.
Amphiphysin is a protein concentrated in neuronal synapses and peripherally associated with neurotransmitter vesicles. It is expressed in many neurons of the central and peripheral nervous systems, in the adrenal medulla, in the anterior and posterior pituitary, in cell lines of the endocrine pancreas, and in spermatocytes. Its subcellular localization and tissue distribution indicate a potential involvement in mechanisms of regulated exocytosis. A role in the dynamic organization of the membrane-associated cytoskeleton is suggested by structural homology to the products of two yeast genes, RVS161 and RVS167, whose mutation results in an abnormal actin distribution, disturbs budding morphology, and impairs cell entry into stationary phase. Limited stretches of sequence similarity, including an SH3 domain, are also shared with other actin-binding proteins. Amphiphysin is the dominant autoantigen in paraneoplastic Stiff-Man syndrome, a neurological autoimmune ...
PURPOSE: Hepatocellular carcinoma (HCC) is a highly virulent malignancy with no effective treatment thus requiring innovative and effective targeted therapies. The oncogene Astrocyte elevated gene-1 (AEG-1) plays a seminal role in hepatocarcinogenesis and profoundly downregulates insulin-like growth factor binding protein-7 (IGFBP7). The present study focuses on analyzing potential tumor suppressor functions of IGFBP7 in HCC and the relevance of IGFBP7 downregulation in mediating AEG-1 function.EXPERIMENTAL DESIGN: IGFBP7 expression was detected by immunohistochemistry in HCC tissue microarray and real-time PCR and ELISA in human HCC cell lines. Dual Fluorescence in situ hybridization was performed to detect loss of heterozygosity at IGFBP7 locus. Stable IGFBP7-overexpressing clones were established in the background of AEG-1-overexpressing human HCC cells and were analyzed for in vitro proliferation and senescence and in vivo tumorigenesis and ...
/sup 125/I-Calmodulin is internalized by isolated rat renal brush border membrane vesicles (BBV) in a time, temperature and calcium dependent manner. Internalization of /sup 125/I-calmodulin into the osmotically sensitive space of BBV was distinguished from binding of the ligand to the outer BBV surface by examining the interaction of ligand and BBV at different medium osmolarities (300-1100 mosm), uptake was inversely proportional to medium osmolarity. Internalized /sup 125/I-calmodulin was intact and Western blots of solubilized BBV with /sup 125/I-calmodulin demonstrated the presence of several calmodulin-binding proteins of 143, 118, 50, 47.5, 46.5 and 35 kilodaltons which could represent potential intravesicular binding sites for the ligand. Heparin and the related glycosaminoglycan heparin sulfate both showed a dose-dependent inhibition (0.5-50 ..mu..g/ml) of /sup 125/I-calmodulin uptake by BBV, but other sulfated and nonsulfated glycosaminoglycans including ...
Cardiac contractile function is dependent on the integrity and function of the sarcolemmal membrane. Swimming exercise training is known to increase cardiac contractile performance. The purpose of the present study was to examine whether a swimming exercise program would alter sarcolemmal enzyme activity, ion flux, and composition in rat hearts. After approximately 11 wk of exercise training, cardiac myosin and actomyosin Ca2+-adenosinetriphosphatase (ATPase) activity was significantly higher in exercised rat hearts than in sedentary control rat hearts. Glycogen content was increased in plantaris and gastrocnemius muscles from exercised animals as was succinic dehydrogenase activity in gastrocnemius muscle of exercised rats in comparison to sedentary rat preparations. Sarcolemmal vesicles were isolated from hearts of exercise-trained and control rats. Sarcolemmal Na+-K+-ATPase and K+-p-nitrophenylphosphatase activities, Na+-Ca2+ exchange, and passive Ca2+ binding ...
This study describes the chemical composition of the essential oil of Artemisia maritima (Am.Oil) and the pharmacological basis for its medicinal use in gut and airways disorders. Twenty five compounds, composing 93.7% of the oil, were identified; among these, chrysanthenyl propionate and elixene were identified for the first time from any Artemisia species. The Am.Oil (0.3?1.0 mg/mL) suppressed spontaneous and high K+ (80 mM)-induced contractions in isolated rabbit jejunum, suggestive of an antispasmodic effect mediated possibly through calcium channel blockade. The calcium channel blockade activity was confirmed when pre-treatment of the tissue with Am.Oil (0.01?0.03 mg/mL) shifted the Ca++ concentration-response curves to the right, similar to verapamil and papaverine. In isolated trach...
It was developed a new formulation of macro aggregates of Samarium 153 ({sup 153} Sm-MH) for the arthropaties treatment. The radio pharmaceutic was prepared by reaction of Samarium 153 chloride (SmCl{sub 3}) in aqueous environment with sodium boron hydride in NaOH 0.5 N. The microscopic analysis shown that the particles have an average size of 4% m (range 1-14 {mu} m). The velocity of sedimentation was 0.008 cm/min with high stability in vitro in human serum. The biological studies in healthy rabbits, shown that the complex is retained inside the articulation still eight days after of the administration of the radiopharmaceutical. Likewise, it is presented the data of absorbed dose in the different target organs, which was determined by thermoluminescent dosimetry (TLD) through the use of a REMCAL phantom (radiation equivalent manikin calibration). (Author)
Neutron energy spectra were measured for two OSURR experimental facilities. The method of multiple foil activation was used with the SAND-II neutron spectrum unfolding code to obtain spectral data for the Central Irradiation Facility and Rabbit tube. Limited data was obtained for the thermal column using only gold foils. Calculations were made of the neutron environment in various locations using the MORSE code. A shift towards higher neutron energies was observed comparing results for LEU and HEU cores. A slight loss in total thermal neutron flux was measured for all positions. Calculational results show reasonable agreement with measured data.
Aim:To examine the electrophysiological effects of sophocarpine on action potentials (AP) and ionic currents of cardiac myocytes and to compare some of these effects with those of amiodarone.Methods:Langendorff perfusion set-up was used in isolated guinea pig heart, and responses to sophocarpine were monitored using electrocardiograph. Conventional microelectrode, voltage clamp technique and perforated patch were employed to record fast response AP (fAP), slow response AP (sAP) and ionic currents in guinea pig papillary muscle or rabbit sinus node cells.Results:Tachyarrhythmia produced by isoprenaline (15 ?mol/L) could be reversed by sophocarpine (300 ?mol/L). Sophocarpine (10 ?mol/L) decreased the amplitude by 4.0%, maximal depolarization velocity (Vmax) of the fAP by 24.4%...
Two lines of mouse tumor cells were shown to be capable of aggregating mouse and rabbit platelets in vitro. This process required higher Mg/sup 2 +/ concentrations than were needed by other commonly used platelet-aggregating agents. Platelet-aggregating activity was also found in tumor cell membrane fragments. This membrane-bound platelet-aggregating material contained protein, lipid, and carbohydrate moieties. The presence of all three appeared to be essential for stimulating platelet aggregation. Destruction of any component abolished its activity. Platelet aggregation induced by tumor cell membrane fragments was associated with a secretory release reaction. In this process, growth-promoting activity for tumor cells was also released from platelets. These results underline the importance of platelets in establishing tumor metastases.
In FY1994, JMTR was in operation during 4 operation cycles with low enriched Uranium(LEU,20%) fuel for irradiation study of nuclear fuels and materials and for radioisotope production. Irradiation studies were carried out using capsules, Oarai Gas Loop-1(OGL-1), Oarai Shroud Facility(OSF-1) and hydraulic rabbits irradiation facilities in support of LWR, FBR, HTTR and thermonuclear reactor. Irradiation studies on blanket materials were intensively carried out. Power ramping tests were carried out and the future program is under consideration. For R and D works, neutron spectrum evaluation technology, re-instrumentation technique for irradiation fuel rod, remote controlled SEM apparatus and examination technique with miniaturized specimens were successfully developed. (author).
Transparency is essential for normal corneal function. Recent studies suggest that corneal cells express high levels of so-called corneal crystallins, such as aldehyde dehydrogenase (ALDH) and transketolase (TKT) that contribute to maintaining cellular transparency. Stromal injury leads to the appearance of repair phenotype keratocytes, the corneal fibroblast and myofibroblast. Previous studies on keratocytes from species such as bovine and rabbit indicate that the transformation from the normal to repair phenotype is accompanied by a loss of corneal crystallin expression, which may be associated with loss of cellular transparency. Here we investigated if a similar loss occurs with human keratocyte repair phenotypes. Human corneal epithelial cells were collected by scraping and keratocytes...
In this study, we measured the effects of different concentrations of paraquat (0,01 mM and 1,0 mM) on the 1-"1"4CO_2 and 6-"1"4CO_2 production of rabbit lung slices and isolated alveolar macrophages, in 20% and 95% oxygen phases respectively. A 95% oxygen phase induced an increase in the 6-"1"4C-glucose oxidation of control lung slices over a 3-hour period, while the increased activity of the pentose pathway over the first 2 hours started to decline during the third hour of incubation. Paraquat (1,0mM) in 20% oxygen caused a consistent increase in the 6-"1"4CO_2 production by lung slices, but in a 95% oxygen phase gradually inhibited the 6-"1"4C-glucose oxidation over a period of 3 hours. The pentose phosphate pathway was highly significantly stimulated by 1,0 mM paraquat in 20% and 95% oxygen over 3 hours. When isolated alveolar macrophages (viability 95%) were incubated in a 20% and 95% oxygen phase respectively, both the 6-"1"4C-glucose and 1-"1"4C-glucose ...
A portion of the active site of rabbit skeletal myosin near the ribose ring of ATP can be labeled by the photoaffinity analogue 3{prime}(2{prime})-O-(4-benzoylbenzoyl)adenosine triphosphate (Bz{sub 2}ATP). The specificity of the photolabeling was assured by first trapping ({sup 14}C)Bz{sub 2}ATP at the active site by use of thiol cross-linking agents. Five radioactive peptides were isolated by high-performance liquid chromatography after extensive trypsin and subtilisin digestion of photolabeled myosin subfragment 1. Four of these peptides were sequenced by Edman techniques, and all originated from a region with the sequence Gly-Glu-Ile-Thr-Val-Pro-Ser-Ile-Asp-Asp-Gln, which corresponds to rabbit myosin heavy chain residues 312-328. The fifth labeled peptide had an amino acid composition appropriate for residues 312-328. Amino acid composition, radiochemical analysis, and sequence data indicate that Ser-324 is the major amino acid residue ...
Aldosterone has been suggested to elicit vessel contraction via a nongenomic mechanism. We tested this proposal in microdissected, perfused rabbit renal afferent arterioles. Aldosterone had no effect on internal diameter in concentrations from 10(-10) to 10(-5) mol/L, but aldosterone abolished the ability of 100 mmol/L KCl to induce vascular contraction. The inhibitory effect of aldosterone was observed from 1 pmol/L. The inhibitory effect was significant after 5 minutes and maximal after 20 minutes and was fully reversible. Actinomycin D (10(-6) mol/L) prolonged the effect of aldosterone. The effect was abolished by the mineralocorticoid receptor antagonist spironolactone (10(-7) mol/L) but not by the glucocorticoid receptor antagonist mifepristone (10(-6) mol/L). The K+-mediated increase of intracellular calcium concentration in afferent arterioles was not affected by aldosterone. Mineralocorticoid receptor was detected by reverse transcription-polymerase chain ...
The authors have shown previously that the domain recognizing receptors on activated human platelets is located on the human fibrinogen {gamma} chain between residues 400 and 411. To study the correlation between the structure of this segment of the {gamma} chain and its reactivity toward receptors on ADP-activated human platelets, they designed a series of analogues containing replacements at 9 out of 12 positions. A double substitution of the normal His{sup 400}-His{sup 401} sequence by Ala-Ala reduced the inhibitory potency of the dodecapeptide 3-fold. When Lys{sup 406} was replaced by Arg, the inhibitory potency of the dodecapeptide decreased 15 times. On the other hand, substitution of Ala{sup 408} with Arg increased the inhibitory potency of the dodecapeptide 6-fold. A drastic decrease in the reactivity of the dodecapeptide toward platelet receptors was observed when Val{sup 411} was replaced by leucine or cysteine or tyrosine. A 3-fold decrease in reactivity was noted when ...
(Materials and Methods) (I) VX/sub 2/ carcinoma was transplanted to the rabbit tibia, sup(99m)Tc-MDP was intravenously injected when bone destruction was radiologically seen. Preoperative scintigraphs, postoperative one of the resected tibia and fibra and scintigraphs of sliced tumor were compared with each other. Tissue fragments were collected from the sliced tumor, and RI was measured to compare concentration rates at different parts together with histological investigations. (II) Tetracycline was given to the rabbit of Experiment (I) to compare with the concentration of sup(99m)Tc-MDP. (III) Microautoradiography method was done on the experimental animals for investigation of the concentration of sup(99m)Tc-MDP. (Results) (I) Localization of sup(99m)Tc-MDP was increased at the sites of tumorous bone formation, of reactive bone formation at tumor edges, and of cartilage calcification, with little concentration at tumor cells or necrotic ...
Tc-MDP 10mCi was intravenously given preoperatively to 20 patients with primary bone tumors and 1 with a metastatic bone tumor. Preoperative and postoperative scintigraphs of resected tumor and scintigraphs of sliced tumor were compared. Tissue fragments were collected from the sliced tumor, and RI measured to compare concentration rates at different areas. Histological investigations were also done. Localization of Tc-MDP was increased at sites of tumorous bone formation, of reactive bone formation at tumor edges, and of cartilage calcification, with little concentration at tumor cells or necrotic tissue. VX/sub 2/ carcinoma was transplanted into the rabbit tibia, Tc-MDP was intravenously injected when bone destruction was radiologically evident, and investigations were done as in the clinical cases. In experimental bone tumor, increased uptake localization of Tc-MDP was much the same as in clinical cases. Tetracycline was given to the rabbit ...
Previous in vitro studies have demonstrated that Ho:YLF laser is capable of inducing physical and chemical changes on dental surfaces treated for caries prevention. The temperature in the pulp chamber was in vitro evaluated to as a function of the power and frequency of the laser irradiation. The purpose of this work is to verify the occurrence of pulp inflammation after Ho:YLF laser irradiation using different parameters in rabbits' teeth. The premolars and molars of ten rabbits (NZB) were divided in two groups according to the irradiation energy values of a Ho:YLF laser prototype operating at 2.065{mu}m wave length, frequency of O,5Hz and pulse length of 250{mu}s. An group A teeth were irradiated with using ten pulses of 334mJ/pulse of a Ho:YLF laser prototype operating at O.5Hz, and group B, with 512mJ/pulse. Animals were killed by transcardiac perfusion and the samples were prepared for histopathological analysis. The in vitro ...
In 65 adult rabbits of both sexes (2.5 to 6 kg) the following salvaging procedures were performed after simulated trauma: (1) Continuous suture line; (2) closure with fibrin sealant after temporary clamping; (3) hemisplenectomy with omental wrap; (4) suture fixation of fragments and slices of spleen tissue to the peritoneum; (5) suture fixation of spleen fragments to the omentum; (6) extraperitoneal subcutaneous transposition of spleen cubes. In addition, the likelihood of spontaneous splenosis was examined in a preliminary study involving 5 animals by introducing unattached spleen meal and fragments. Scanning was repeated 4 to 12 weeks postoperatively. The functional competence of minor parenchymal islands was also investigated with well counter measurements of organ and muscle tissue homogenates. Spontaneous splenosis after abdominal injuries was not found to be the rule. Salvaging procedures at the natural vascular hilus by sutures, fibrin sealant and ...
The affinity label 8-((4-bromo-2,3-dioxobutyl)thio)adenosine 5{prime}-triphosphate (8-BDB-TA-5{prime}-TP) reacts covalently with rabbit muscle pyruvate kinase, incorporating 2 mol of reagent/mol of enzyme subunit upon complete inactivation. Protection against inactivation is provided by phosphoenolpyruvate, K{sup +}, and Mn{sup 2+} and only 1 mol of reagent/mol of subunit is incorporated. The authors have now identified the resultant modified residues. After reaction with 8-BDB-TA-5{prime}-TP at pH 7.0, modified enzyme was incubated with ({sup 3}H)NaBH{sub 4} to reduce the carbonyl groups of enzyme-bound 8-BDB-TA-5{prime}-TP and to introduce a radioactive tracer into the modified residues. Following carboxymethylation and digestion with trypsin, the radioactive peptides were separated on a phenylboronate agarose column followed by reverse-phase high-performance liquid chromatography in 0.1% trifluoroacetic acid with an acetonitrile gradient. Gas-phase sequencing ...
We have done analysis of elements in the Baltic sea sediment by using gamma spectrometry in neutron activation analysis Goal of the analysis is to determine the rate of morsel elements which is contained in the Baltic sea sediment. Sample of the Baltic sea Sediment (IAEA-SED-1), Standard of Reference Material (SRM) 1646 Estuarine Sediment and Standard Ce, Fe, Sc, and Co are put into vial of capsule rabbit, then irradiated together for 30 minutes in rabbit facility Multi Purpose Reactor at power of 25 MW and then cooled for one week. Next sample, standard, and SRM which have irradiated, each is cut for 30 minutes using gamma spectrometer. Result of analysis of rate of elements Ce, Fe, Sc and Co in the baltic sea sediment successively equal to 141.20 ppm, 5.91 %, 14.88 ppm, and 19.59 ppm. Result of analysis in SRM Estuarine sediment successively equal to 96.60 ppm, 3.21 %, 10.25 ppm, and 9.63 ppm. Result of this analysis if compared to the value ...
Human complement protein C9 is shown to be a metalloprotein that binds 1 mol of Ca"2"+/mol of C9 with a dissociation constant of 3 #mu#m as measured by equilibrium dialysis. Incubation with EDTA removes the bound calcium, resulting in a apoprotein with decreased thermal stability. This loss in stability leads to aggregation and, therefore, to loss of hemolytic activity upon heating to a few degrees above the physiological temperature. Heat-induced aggregation of apoC9 can be prevented by salts that stabilize proteins according to the Hofmeister series of lyotropic ions, suggesting that the ion in native C9 may ligand with more than one structural element of domain of the protein. Ligand blotting indicates that the calcium binding site is located in the amino-terminal half of the protein. Removal of calcium by inclusion of EDTA in assay mixtures has no effect on the hemolytic activity of C9, and its capacity to bind to C8 in solution, or to small unilamellar lipid ...
Effects of ethanol in vitro on membrane vesicles were evaluated by monitoring "3"6Cl"- influx. Different assay parameters were tested to determine increased or decreased action of ethanol on GABA-activated chloride channels. The ability of 30 mM ethanol to augment "3"6Cl"- flux was seen at 0 degree C, in the absence of GABA, and at 34 degree C in the presence of GABA, using two different assay procedures. Picrotoxin blocked the direct effects of ethanol suggesting GABA_a involvement. Endogenous GABA in the medium surrounding the microsacs was assayed at different temperatures both in the presence and absence of GABA and ethanol. The direct effect of ethanol did not appear to involve the action of endogenous GABA. In addition to temperature effects on the assay, time of membrane storage also influenced ethanol action. Microsacs stored on ice for 2 hours or more lost their ability to respond to ethanol but not to GABA, pentobarbital or flunitrazepam. When these drugs ...
This project is concerned with the structure and function of the unique antenna system found in the green photosynthetic bacteria. The antenna system in these organisms is contained within a vesicle known as a chlorosome, which is attached to the cytoplasmic side of the cell membrane. Additional antenna pigments and reaction centers are contained in integral membrane proteins. Energy absorbed by the bacteriochlorophyll c (BChl c) pigments in the chlorosome is transferred via a ``baseplate`` array of BChl a antenna pigments into the membrane and to the reaction center. This system is similar in some respects to the phycobilisome antenna system found in cyanobacteria and some types of algae, in that a membrane-associated structure absorbs light and transfers it to the membrane where conversion to chemical energy takes place. However, the overall structure, the type of pigments utilized and the nature of the proteins in these two types of membrane-associated antenna ...
This study aims to investigate the difference in the interaction of antimicrobial peptides with two classes of zwitterionic peptides, phosphatidylethanolamines (PE) and phosphatidylcholines (PC). Further experiments were performed on model membranes prepared from specific bacterial lipids, lipopolysaccharides (LPS) isolated from Salmonella minnesota. The structure of the lipid-peptide aqueous dispersions was studied by small-and wide-angle X-ray diffraction during heating and cooling from 5 to 85 C. The lipids and peptides were mixed at lipid-to-peptide ratios 10-10000 (POPE and POPC) or 2-50 (LPS). All experiments were performed at synchrotron soft condensed matter beamline A2 in Hasylab at Desy in Hamburg, Germany. The phases were identified and the lattice parameters were calculated. Alamethicin and melittin interact in similar ways with the lipids. Pure POPC forms only lamellar phases. POPE forms lamellar phases at low temperatures that upon heating transform into a highly curved ...
Active oxygens produced by low-dose irradiation can induce the synthesis of proteins involved in the active oxygen reduction path and can activate the chemical self-defense mechanisms in vivo, which can alleviate the injuries caused by active oxygens themselves. The following findings suggests that it is not only by low-dose irradiation that can induce the responses described above but also by any physical and/or chemical stresses which can produce small amount of active oxygens in vivo. 1. X-irradiation with 0.5 Gy and/or water immersion and restraint induced heat shock protein (HSP)70 in rat stomach cells. HSP70 was also induced in adrenal cells by paraquat administration. 2. The SOD activity in pancreas of rat significantly increased by #gamma#-irradiation with 0.5 Gy. 0.5 Gy irradiation preceeding the alloxan administration significantly suppressed the increase in pancreatic lipid peroxides and in blood glucose levels, degranulation in #beta# cells, and decrease in blood insulin ...
The effects of 0.2 and 0.5 mL/kg Innovar (a neuroleptic analgesic) on cardiovascular functions and reflexes in rabbits were measured. We recorded the effects of Innovar on arterial pressure, heart rate, respiration rate, ventricular contractility, arterial oxygen tension, as well as the drug's effects on the bradycardia and vasoconstrictor response to cigarette smoke stimulation of the nose (the so-called "nasopharyngeal reflex"). In animals given 0.2 mL/kg Innovar, all steady state cardiovascular variables had returned to pre-Innovar levels in 45 min, as had the efficacy of the nasopharyngeal reflex. In animals given 0.5 mL/kg Innovar, all steady state cardiovascular variables, except PaO2, were slightly but significantly depressed for up to 135 min after injection. The nasopharyngeal reflex returned to normal within 90 min. Because of the calmative and analgesic effects of Innovar, and its only moderate effects on cardiovascular functions and reflexes, we feel it ...
The Ohio State University Research Reactor (OSURR) is licensed to operate at a maximum power level of 500 kW. A pool-type reactor using flat-plate, low enriched fuel elements, the OSURR provides several experimental facilities including two 6-inch i.d. beam ports, a graphite thermal column, several graphite-isotope-irradiation elements, a pneumatic transfer system (Rabbit), various dry tubes, and a Central Irradiation Facility (CIF). The core arrangement and accessibility facilitates research programs involving material activation or core parameter studies. The OSURR control room is large enough to accommodate laboratory groups which can use control instrumentation for monitoring of experiments. The control instrumentation is relatively simple, without a large amount of duplication. This facilitates opportunities for hands-on experience in reactor operation by nuclear engineering students making reactor parameter measurements. For neutron activation analysis and ...
Intramuscular hermorrhage was induced by injecting autologous blood into the paraspinal muscle of 8 rabbits. In order to evaluate the time-dependent changes of hemorrhage observed on MRI, the animals were imaged at different stages of blood resolution at 0.02 tesla (T), and control examined with ultrasound using a 7.5 MHz linear transducer. Six inversion recovery sequences (TR=1 000 ms. TE=30 ms, and TI=18, 48, 148, 201, 302, and 398 ms) were used for the invivo calculation of T1 relaxation times. IR 1 000 (398)/30 imaging was performed before and after the Gd-DOTA administration. The hemorrhage was evident on MR images throughout the study, especially on the T2 weighted (SE 1 000/100) images. MRI showed the healing lesion longer than ultrasound. The T1 relaxation time increased during the time of resolution. Lesions on days 4 to 7 enhanced in intensity after the injection of Gd-DOTA. (orig.).
A tick/rickettsial survey in various parts of Switzerland revealed the presence of a new, hitherto undescribed spotted fever group rickettsia ("Swiss agent") in up to 11.7% of I. ricinus collected off vegetation. Infection in ticks was found to be generalized with rickettsiae developing intracellularly and occasionally also intranuclearly. As a result of massive growth in ovarial tissues, including the germinative cells, the rate of transovarial and filial infection was 100%. The "Swiss agent" appears to be nonpathogenic for guinea pigs, domestic rabbits, and Swiss mice, but in male meadow voles (Microtus pennsylvanicus) it produces a microscopically detectable infection in the tunica vaginalis. The rickettsia grows well in tissue culture systems including chick embryo fibroblast, Vero, and vole tissue cells, when inoculated via yolk sac into 5-day-old hens' eggs, it kills 100% of the embryos after 5 to 7 days. Antigenic relatedness of the "Swiss agent" to ...
The heavy chains of Acanthamoeba myosins, IA, IB and II, turkey gizzard myosin, and rabbit skeletal muscle myosin subfragment-1 were specifically labeled by radioactive ATP, ADP, and UTP, each of which is a substrate or product of myosin ATPase activity, when irradiated with uv light at 0"0C. With UTP, as much as 0.45 mol/mol of Acanthamoeba myosin IA heavy chain and 1 mol/mol of turkey gizzard myosin heavy chain was incorporated. Evidence that the ligands were associated with the catalytic site included the observations that reaction occurred only with nucleotides that are substrates or products of the ATPase activity; that the reaction was blocked by pyrophosphate which is an inhibitor of the ATPase activity; that ATP was bound as ADP; and that label was probably restricted to a single peptide following limited subtilisin proteolysis of labeled Acanthamoeba myosin IA heavy chain and extensive cleavage with CNBr and trypsin of labeled turkey gizzard myosin heavy ...
Treatment of cultures of 3T3-L1 cells with methylisobutyl-xanthine and dexamethasone has been shown to result in accumulation of lipid and conversion to the morphology of adipocytes in more than 90% of the cells. The status of the stimulatory (Gs), inhibitory (Gi) and Go-proteins during the course of 3T3-L1 differentiation was examined. The amount of alpha subunit of Gs (#alpha#Gs), assayed by radiolabeling in the presence of cholera toxin and ["3"2P]NAD"+, increased upon differentiation as previously described by others. The amounts of #alpha#Gi and #alpha#Go assayed by radiolabeling in the presence of pertussis toxin and ["3"2P]NAD"+ increased 3-fold upon differentiation. Immunoblots of cell membranes subjected to gel electrophoresis in sodium dodecyl sulfate were probed with two rabbit antisera raised against bovine brain #alpha#Go and with one raised against the#beta#-subunit of the bovine rod-outer-segment G-protein, referred to as transducin. The ...
In 18 piglets, weighing 10-15 kg, third-degree burns or full-thickness skin excisions of 4 X 4 cm were inflicted. The effect of five dressing materials on adhesiveness to the wounds, appearance, conformability, wound contraction, bacterial count, and morphology of the wound was studied at the end of the seventh and fourteenth days without dressing changes. In 11 piglets with a burn wound, the most adherent dressing was collagen sponge(CS), followed by polyurethane sponge (PU), pigskin xenograph (PS), and xeroform. CS more effectively debrided the wound from coagulated necrotic tissue than the other dressings. Wound contraction was maximal with CS dressing (52%), followed by PU (44%), xeroform (32%), and PS (27%). In another seven piglets with full-thickness excised wounds, a velour dressing made of polytetrafluoroethylene (PTFE) or PU adhered significantly more than CS or PS. Wound contraction was greater with CS (37%) or PTFE (35%) than with PS (23%) or PU (18%). The bacterial counts ...
Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods. In one, a human cDNA library in phage {lambda} was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a {lambda}gt11 expression vector was screened for the expression of antigenic determinants that are recognized by rabbit antibodies specific to human DNA topoisomerase II. The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is encoded by a single-copy gene. The location of the gene was mapped to chromosome 17q21-22 by in situ hybridization of a cloned fragment to ...
The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on pyrimidine ring saturation products (thymine glycol.and ...
The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on pyrimidine ring saturation products (thymine glycol.and ...
Autophagy is an intracellular recycling route in eukaryotes whereby organelles and cytoplasm are sequestered in vesicles, which are subsequently delivered to the vacuole for breakdown. The process is induced by various nutrient-responsive signaling cascades converging on the Autophagy-Related1 (ATG1)/ATG13 kinase complex. Here, we describe the ATG1/13 complex in Arabidopsis thaliana and show that it is both a regulator and a target of autophagy. Plants missing ATG13 are hypersensitive to nutrient limitations and senesce prematurely similar to mutants lacking other components of the ATG system. Synthesis of the ATG12-ATG5 and ATG8-phosphatidylethanolamine adducts, which are essential for autophagy, still occurs in ATG13-deficient plants, but the biogenesis of ATG8-decorated autophagic bodies does not, indicating that the complex regulates downstream events required for autophagosome enclosure and/or vacuolar delivery. Surprisingly, levels of the ATG1a and ATG13a ...
In APCs, presentation by MHC II molecules of the chemically dominant peptide from the protein hen egg white lysozyme (HEL) generates different conformational isomers of the peptide-MHC II complexes (pMHC). Type B pMHCs are formed in early endosomes from exogenous peptides in the absence of H2-DM, whereas in contrast, type A pMHC complexes are formed from HEL protein in late vesicles after editing by H2-DM. Thus, H2-DM edits off the more unstable pMHC complexes, which are not presented from HEL. In this study, we show that type B pMHC complexes were presented from HEL protein only after stimulation of dendritic cells (DC) with TLR ligands or type I IFN. Type I IFN contributed to most TLR ligand-induced type B pMHC generation, as presentation decreased in DC lacking the receptor for type I IFNs (IFNAR1(-/-)). In contrast, presentation of type A pMHC from HEL and from peptide was minimally affected by TLR ligands. The relative effectiveness of CD8?(+) DC or CD8?(-) DC ...
Pancreatic amyloid deposits of amylin are a hallmark of Type II diabetes and considerable evidence indicates that amylin oligomers are cytotoxic to beta-cells. Many efforts are presently spent to find out naturally occurring molecules, or to design synthetic ones, able to hinder amylin aggregation or to protect cells against aggregate cytotoxicity. In this context, a protective effect of some polyphenols against amyloid cytotoxicity was reported. Actually dietary polyphenols are endowed with multiple health benefits, and extra virgin olive oil is attracting increasing interest as a source of these substances. Here, we investigated the effects on amylin aggregation and cytotoxicity of the secoiridoid oleuropein aglycon, the main phenolic component of extra virgin olive oil. We found that oleuropein, when present during the aggregation of amylin, consistently prevented its cytotoxicity to RIN-5F pancreatic beta-cells, as determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl ...
Asialoorosomucoid (ASOR) is internalized and degraded by HepG2 cells after binding to the asialoglycoprotein (ASGP) receptor, internalization through the coated pit/coated vesicle pathway, and trafficking to lysosomes. Primaquine, an 8-aminoquinoline antimalarial compound, inhibits ASOR degradation at concentrations greater than 0.2 mM by neutralizing intracellular acid compartments. This leads to alterations in surface receptor number, receptor-ligand dissociation, and receptor recycling. We have investigated the effects of primaquine on 125I-ASOR uptake and degradation as a function of primaquine concentration and duration of exposure. Concentrations below those required for neutralization of acidic compartments block 125I-ASOR degradation in HepG2 cells and lead to intracellular ligand accumulation. This effect is maximal at 80 microM primaquine. The intracellular 125I-ASOR is undegraded, dissociated from the ASGP receptor, and contained within vesicular ...
Asialoorosomucoid (ASOR) is internalized and degraded by HepG2 cells after binding to the asialoglycoprotein (ASGP) receptor, internalization through the coated pit/coated vesicle pathway, and trafficking to lysosomes. Primaquine, an 8-aminoquinoline antimalarial compound, inhibits ASOR degradation at concentrations greater than 0.2 mM by neutralizing intracellular acid compartments. This leads to alterations in surface receptor number, receptor-ligand dissociation, and receptor recycling. We have investigated the effects of primaquine on 125I-ASOR uptake and degradation as a function of primaquine concentration and duration of exposure. Concentrations below those required for neutralization of acidic compartments block 125I-ASOR degradation in HepG2 cells and lead to intracellular ligand accumulation. This effect is maximal at 80 microM primaquine. The intracellular 125I-ASOR is undegraded, dissociated from the ASGP receptor, and contained within vesicular ...
Although the intracellular ultrastructural processing of epidermal growth factor (EGF) and its receptor have been described in cell culture systems, very few studies have examined this phenomenon in intact tissues. We have examined the ultrastructural and biochemical handling of ["1"2"5I]EGF in the epidermis and hair follicle bulb of intact, viable, 3- to 5-day-old rat skin the EGF receptor distribution of which has already been documented and in which EGF has been shown to be biologically active. After incubation of explants with 10 nM ["1"2"5I]EGF for 2.5 h at 25 degrees or 37 degrees C, radiolabel was detected over the basal cells of the epidermis and hair follicle outer root sheath, confirming previous light microscope observations. More specifically, silver grains were observed near coated and uncoated plasma membrane and coated membrane invaginations, Golgi apparatus, lysosomal structures, and nuclei. Sodium azide inhibited internalization of label, whereas a series of lysosomal ...
Although the intracellular ultrastructural processing of epidermal growth factor (EGF) and its receptor have been described in cell culture systems, very few studies have examined this phenomenon in intact tissues. We have examined the ultrastructural and biochemical handling of (/sup 125/I)EGF in the epidermis and hair follicle bulb of intact, viable, 3- to 5-day-old rat skin the EGF receptor distribution of which has already been documented and in which EGF has been shown to be biologically active. After incubation of explants with 10 nM (/sup 125/I)EGF for 2.5 h at 25 degrees or 37 degrees C, radiolabel was detected over the basal cells of the epidermis and hair follicle outer root sheath, confirming previous light microscope observations. More specifically, silver grains were observed near coated and uncoated plasma membrane and coated membrane invaginations, Golgi apparatus, lysosomal structures, and nuclei. Sodium azide inhibited internalization of label, whereas a series of ...
The synapsin III gene, SYN3, which belongs to the family of synaptic vesicle-associated proteins, has been implicated in the modulation of neurotransmitter release and in synaptogenesis, suggesting a potential role in several neuropsychiatric diseases. The human SYN3 gene is located on chromosome 22q12-13, a candidate region implicated in previous linkage studies of schizophrenia. However, association studies of SYN3 and schizophrenia have produced inconsistent results. In this study, four SYN3 SNPs (rs133945 (-631 C>G), rs133946 (-196 G>A), rs9862 and rs1056484) were tested in three sets of totally 3759 samples that comprise 655 affected subjects and 626 controls in the Irish Case-Control Study of Schizophrenia (ICCSS), 1350 samples incorporating 273 pedigrees in the Irish Study of High Density Schizophrenia Families (ISHDSF), and 564 unrelated schizophrenia patients and 564 healthy individuals in a Chinese case-control sample. The expression levels of SYN3 ...
A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. ...
A #lambda#gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. ...
A comparison of the concentration-response effects of inhaled ozone (O/sub 3/) in different species of laboratory animals was made in order to better understand the influence of the choice of species in inhalation studies of this gas. The effect of 4-hour exposure to ozone (O/sub 3/) at concentrations of 0.0, 0.2, 0.5, 1.0, and 2.0 ppm was determined in rabbits, guinea pigs, rats, hamsters and mice. Lavage fluid protein (LFP) accumulation 18 hr after exposure was used as the indicator of O/sub 3/-induced pulmonary edema. All species had similar basal levels of LFP (250-350 ug/ml) when a volume of saline which approximated the total lung capacity was used for lavage of the collapsed lungs. Exponential dose-response curves were seen in all species except guinea pigs, which showed significant increases in LFP at low O/sub 3/ concentrations (0.2 ppm) and a leveling off of response at the higher O/sub 3/ levels. Other species usually showed significant elevations in LFP ...
The authors have localized and characterized 2-125I-iodomelatonin binding sites in the chicken brain using in vitro quantitative autoradiography. Binding sites were widely distributed throughout the chicken brain, predominantly in regions associated with the visual system. The specific binding of 2-125I-iodomelatonin to discrete chicken brain areas was found to be saturable, reversible, and of high affinity. The specific binding of 2-125I-iodomelatonin (75 pm) was quantitated for 40 identifiable brain regions. Eight brain regions were chosen for binding characterization and pharmacological analysis: optic tectum, Edinger-Westphal nucleus, oculomotor nucleus, nucleus rotundus, ventral supraoptic decussation, ventrolateral geniculate nucleus, neostriatum, and ectostriatum. These regions showed no rostral-caudal gradient in 2-125I-iodomelatonin specific binding, and saturation analysis revealed a single class of high-affinity sites with KD values in the range of 33-48 pM and receptor site ...
Previous investigations have shown that both the early biological response and the mechanical properties of a porous hydroxyapatite bone graft substitute are highly sensitive to its pore structure. The objective of this study was to evaluate whether the pore structure continued to influence bone integration in the medium to long term. Two screened batches of porous hydroxyapatite (PHA) designated as batch A and batch B, with porosities of approximately 60 and 80%, respectively, were selected for this study and implanted for periods of 5, 13, and 26 weeks into the lower femur of New Zealand White rabbits. Histomorphometric analysis of the absolute volume of bone ingrowth within batch A and B implants from 5 to 26 weeks showed that the absolute volume of bone ingrowth was consistently lower in batch A (10-21%), compared to batch B implants (24-31%). However, when the volume of bone ingrowth was normalised for the available pore space, this difference was reduced ...
Rabbit antisera raised to human and chicken MHC molecules were used to immunoprecipitate cross-reactive molecules from biosynthetically and cell surface-labeled spleen and/or blood cells of representative vertebrate species. Five major points emerged: 1) There were many nonspecific cross-reactions using these techniques, so various criteria were developed to distinguish these from true MHC-like molecules. 2) Only very small subpopulations of immunogen-specific antibodies cross-reacted with MHC-like molecules in other nonmammalian species. These subpopulations were different for each species and even within a species, sometimes being so limited as to behave like alloantisera. This led to a very scattered pattern of true cross-reactions that sometimes failed to reflect the properties of the bulk antibody population. 3) Antisera containing antibodies to class II beta- and class I alpha-chains cross-reacted better and more widely than those to B-G, class II alpha and, ...
We present a femtosecond Laser Two-Photon Polymerization (LTPP) system of large scale three-dimensional structuring for applications in tissue engineering. The direct laser writing system enables fabrication of artificial polymeric scaffolds over a large area (up to cm in lateral size) with sub-micrometer resolution which could find practical applications in biomedicine and surgery. Yb:KGW femtosecond laser oscillator (Pharos, Light Conversion. Co. Ltd.) is used as an irradiation source (75 fs, 515 nm (frequency doubled), 80 MHz). The sample is mounted on wide range linear motor driven stages having 10 nm sample positioning resolution (XY--ALS130-100, Z--ALS130-50, Aerotech, Inc.). These stages guarantee an overall travelling range of 100 mm into X and Y directions and 50 mm in Z direction and support the linear scanning speed up to 300 mm/s. By moving the sample three-dimensionally the position of laser focus in the photopolymer is changed and one is able to write complex 3D ...
The purpose was to develop a simplified and reliable method of separating free from antibody-bound ligand using a precipitating antibody linked to a cellulose derivative. Dose-response curves and control sera were set up in parallel for various pituitary and placental polypeptides, steroid hormones, insulin, glucagon, triiodothyronine, thyroxine, angiotensin I, calcitonin, gastrin, cyclic AMP, and digoxin. After first-antibody reactions had reached equilibrium, free and bound ligand were separated using a double-antibody solid-phase system in parallel with conventional methods, including dextran-coated charcoal, double-antibody precipitation, single-antibody solid phase, organic solvents, salt precipitation, and anion-exchange resins. The effect of variations in temperature, incubation time, protein content, pH, and amount of separating material added were studied. The results showed that separation was complete within 1 hr for small ligand molecules and within 2 hr for larger ones. ...
The purpose was to develop a simplified and reliable method of separating free from antibody-bound ligand using a precipitating antibody linked to a cellulose derivative. Dose-response curves and control sera were set up in parallel for various pituitary and placental polypeptides, steroid hormones, insulin, glucagon, triiodothyronine, thyroxine, angiotensin I, calcitonin, gastrin, cyclic AMP, and digoxin. After first-antibody reactions had reached equilibrium, free and bound ligand were separated using a double-antibody solid-phase system in parallel with conventional methods, including dextran-coated charcoal, double-antibody precipitation, single-antibody solid phase, organic solvents, salt precipitation, and anion-exchange resins. The effect of variations in temperature, incubation time, protein content, pH, and amount of separating material added were studied. The results showed that separation was complete within 1 hr for small ligand molecules and within 2 hr for larger ones. ...
Duchenne and Becker muscular dystrophies are caused by defects of dystrophin, which forms a part of the membrane cytoskeleton of specialized cells such as muscle. It has been previously shown that the dystrophin-associated protein A1 (59-kDa DAP) is actually a heterogeneous group of phosphorylated proteins consisting of an acidic ({alpha}-A1) and a distinct basic ({beta}-A1) component. Partial peptide sequence of the A1 complex purified from rabbit muscle permitted the design of oligonucleotide probes that were used to isolate a cDNA for one human isoform of A1. This cDNA encodes a basic A1 isoform that is distinct from the recently described syntrophins in Torpedo and mouse and is expressed in many tissues with at least five distinct mRNA species of 5.9, 4.8, 4.3, 3.1, and 1.5 kb. A comparison of the human cDNA sequence with the GenBank expressed sequence tag (EST) data base has identified a relative from human skeletal muscle, EST25263, which is probably a human ...
A burgeoning interest in technetium coordination chemistry has been spurred by its widespread use in nuclear medicine. Recent focus has been on the design and preparation of new technetium complexes, the lipophilicity, polarity, and overall charge of which can be readily altered by simple substitutions in the molecular framework of the ligand in order to optimize the biodistribution and target specific organs. Examples are cationic (Cardiolite) and neutral (Ceretec) technetium complexes, used as myocardial and brain imaging agents, respectively. The authors have been pursuing a continuing study of tris(3-hydroxy-4-pyridinonato)gallium and -indium complexes that are water soluble, are hydrolytically stable, and are of varying lipophilicity and neutral charge. Studies of in vivo in rabbits and mice have revealed that several of these cationic complexes may be useful as morphologic kidney imaging agents, with [{sup 99m}Tc(pap){sub 3}]{sup +} showing elevated levels in ...
A burgeoning interest in technetium coordination chemistry has been spurred by its widespread use in nuclear medicine. Recent focus has been on the design and preparation of new technetium complexes, the lipophilicity, polarity, and overall charge of which can be readily altered by simple substitutions in the molecular framework of the ligand in order to optimize the biodistribution and target specific organs. Examples are cationic (Cardiolite) and neutral (Ceretec) technetium complexes, used as myocardial and brain imaging agents, respectively. The authors have been pursuing a continuing study of tris(3-hydroxy-4-pyridinonato)gallium and -indium complexes that are water soluble, are hydrolytically stable, and are of varying lipophilicity and neutral charge. Studies of in vivo in rabbits and mice have revealed that several of these cationic complexes may be useful as morphologic kidney imaging agents, with ["9"9"mTc(pap)_3]"+ showing elevated levels in the kidneys ...
Intact rat fat cell exposed to 12.5 ..mu..M (..gamma..-32P)ATP incorporate label into specific proteins within minutes. By solubilizing the reaction mixture with SDS which bypasses the subcellular fractionation steps, the labeled proteins can be identified in autoradiographs of SDS-PAGE gels. The most prominently labeled protein has an M/sub r/ of 42,000. Localization of this component to the cell surface can be made on the basis of inhibition of phosphorylation by addition of a protein derived from the rat brain with protein kinase inhibitory property, susceptibility of the phosphorylated protein to the tryptic digestion, inhibition of phosphorylation of this protein after brief exposure to melittin. To rule out the possibility that the cell surface protein might be a mitochondrial contaminant from broken cells, /sup 32/Pi-labeled and (..gamma..-/sup 32/P)ATP-labeled cells were chromatographed on a rabbit anti-pyruvate dehydrogenase antibody-Sepharose 4B column. A ...
Glycosaminoglycans (GAG) are believed to be important in the pathogenesis of atherosclerosis. We have previously demonstrated that areas of injured aorta that have been re-endothelialized accumulate increased amounts of lipid and GAG when compared to areas remaining de-endothelialized. We have now examined the net incorporation of labeled precursors into the individual GAG present in both re-endothelialized and de-endothelialized areas of rabbit aorta. Aortic tissue was examined at 2-3 and 10-14 weeks after a denuding injury by incubating tissue minces with (/sup 3/H)glucosamine and sodium (/sup 35/S)sulfate for 24 hr. Following incubation, the aortic GAG were isolated and assayed for uronic acid concentration and radioactivity. Results indicate that the total GAG concentration was significantly greater in the re-endothelialized as compared to de-endothelialized areas. The concentration in uninjured aorta was 9.01. The difference between the injured tissues was ...
The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and ...
Environmental monitoring has been conducted at the U.S. Department of Energy`s (DOE`s) Hanford Site in southeastern Washington State since 1945. Fish and wildlife have been monitored since 1945, however, a major emphasis on mammals did not occur until the 1970s. This report focuses on the 10-year period from 1983 through 1992. The objectives of this report are to evaluate {sup 90}Sr and {sup 137}Cs concentrations in Site wildlife populations and, when possible, evaluate trends in concentrations over this period of time. No distinct trends in radionuclide concentrations were apparent in most species sampled. Many measurements were at or below the analytical limit of detection. This evaluation found that concentrations of {sup 90}Sr in rabbit and deer bone were elevated in animals collected from areas adjacent to industrialized areas. Similarly, radionuclide concentrations in duck muscle from waterfowl collected at B Pond were elevated with {sup 137}Cs when compared ...
Aim: Rheumatoid arthritis is a limiting disease having, among its pathological features, the inflammation of synovial tissue with progressive and later destruction of the articulation. This leads to joint deformation and loss of its function, generating pain and reducing the mobility of the affected articulation. The aim was to evaluate "1"6"6Ho-Hydroxyapatite ("1"6"6 Ho-HA) as potential radiopharmaceutical for the symptomatic treatment of chronic and acute arthritis. Materials and Methods: Holmiun-166 was produced by irradiation of Ho_2O_3 at La Reina Research Reactor, Nuclear Chilean Energy Commission. Hydroxyapatite was in-house synthesized. Its labelling and quality controls follows the internationally accepted procedures. An antigen's arthritis was induced to eight New Zealand rabbits with the "1"6"6 Ho-HA radiochemical being administered thereafter in two dosage modalities (single and double). The compound therapeutic efficiency was evaluated based upon ...
The 2- and 8-azido trimer 5'-triphosphate photoprobes of 2-5A have been enzymatically synthesized from [#gamma#-"3"2P]2-azidoATP and [#alpha#-"3"2P]8-azidoAPT by 2-5A synthetase from rabbit reticulocyte lysates. Identification and structural determination of the 2- and 8-azido adenylate trimer 5'-triphosphates were accomplished by enzymatic hydrolyses with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase. Hydrolysis products were identified by HPLC and PEI-cellulose TLC analyses. The 8-azido photoprobe of 2-5A displaces p_3A_4["3"2P]pCp from RNase L with affinity equivalent to p_3A_3. The 8-azido photoprobe also activates RNase L to hydrolyze poly(U)["3"2P]pCp 50% at 7 /times/ 10/sup /minus/9/ M in core-cellulose assays. The 2- and 8-azido photoprobes and authentic p_3A_3 activate RNase L to cleave 28S and 18S rRNA to specific cleavage products at 10/sup /minus/9/ M in rRNA cleavage assays. The nucleotide binding site(s) of RNase L and/or ...
Thirty-seven patients with primary nonresectable intrahepatic cholangiocarcinoma (57% with prior treatment and/or metastasis) were prospectively treated with external radiation, chemotherapy, and "1"3"1I labelled anti-CEA. Therapy began in all trials with whole liver irradiation (21.0 Gy, 3.0 Gy/Fx, 4 days/week, 10 MV photons) with alternate treatment day chemotherapy (Adriamycin, 15 mg + 5-FU, 500 mg). One month after external beam therapy, chemotherapy was given (Adriamycin, 15 mg + 5-FU, 500 mg) followed the next day by the first administration of "1"3"1I anti-CEA. The treatment schedule used was 20 mCi day 0; 10 mCi day 5 as an outpatient. This schedule was derived from tumor dose estimates which indicated that 20 mCi (8-10 mCi/mg IgG) was sufficient to achieve tumor saturation with a tumor effective half-life of 3 to 5 days, depending upon the species of animal from which the antibody was obtained. The median tumor dose for the 20 mCi + 10 mCi regimen was 6.2 Gy. Antibody therapy ...