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Biomedical subjects

L Li

Publications and source records attributed to L Li.

At least 109 records · Page 6Linked to original sources

Regulation of antigen-specific versus by-stander IgE production after antigen sensitization.

IgE is critical in the pathogenesis of allergic disorders. In this report, we investigated the differential regulation of antigen-specific and by-stander IgE. Ovalbumin (OVA) immunization did not increase IgE producing cells in the spleen, but significantly enhanced the intracellular IgE content of all IgE+ cells. In contrast, OVA induced a significant increase of IgE+ cells in the draining lymph nodes (LN). Furthermore, OVA-specific IgE was detected only in the ex vivo cultures of the draining LN but not the spleen cells, while total IgE was increased in both cultures. These results indicated that antigen-specific IgE was mainly produced in the draining LN, while the spleen was a major source for by-stander IgE. Anti-IL-4, but not anti-IL-13, antibody blocked the expansion of IgE producing cells in the draining LN as well as systemic OVA-specific and total IgE levels, indicating IL-4 was important in both antigen-specific IgE generation and total IgE upregulation.

Adjuvants, Immunologic↗

Receptor mechanisms mediating cyanide generation in PC12 cells and rat brain.

Cyanide is generated in neurons and this report examines the two different receptors which mediate cyanide formation in neuronal tissue. An opiate receptor blocked by naloxone increases cyanide production both in rat brain and in rat pheochromocytoma (PC12) cells. A muscarinic receptor in PC12 cells releases cyanide and the effect is blocked by atropine. In rat brain, in vivo, a muscarinic agonist inhibits cyanide generation, possibly by acting on receptor subtypes different from those in PC12 cells. Cyanide generation by a muscarinic agonist in PC12 cells is blocked by pertussis toxin but that caused by an opiate is not. Thus, two different receptors and two different second messenger systems can mediate cyanide generation in PC12 cells. In parallel with the in vivo data, cultured primary rat cortical cells also show decreased cyanide release following muscarinic stimulation. Both blockade of cyanide generation by muscarinic receptor activation and cyanide release by opiate agonists from cortical cells are pertussis toxin insensitive. Similarly, little cyanide generation was seen following cholera toxin treatment. These data indicate that opiate receptors increase and muscarinic receptors decrease cyanide production in rat brain tissue by G-protein independent mechanisms. This work supports the suggestion that the powerful actions of cyanide may be important for neuromodulation in the CNS.

Analysis of Variance↗

Kynurenate in the pontine reticular formation inhibits acoustic and trigeminal nucleus-evoked startle, but not vestibular nucleus-evoked startle.

The startle reflex is elicited by acoustic, trigeminal or vestibular stimulation, or by combinations of these stimuli. Acoustic startle is mediated largely by ibotenate-sensitive neurons in the ventrocaudal pontine reticular formation (PnC). In these studies we tested whether startle elicited by stimulation of different modalities is affected by infusion of the non-selective glutamate antagonist, kynurenate, into the PnC. In awake rats, startle responses evoked by either acoustic or spinal trigeminal nucleus stimulation were inhibited by kynurenate, but not saline, infusions, with the most effective placements nearest PnC. In chloral hydrate-anesthetized rats, kynurenate in the PnC reduced trigeminal nucleus-evoked hindlimb EMG responses, but not vestibular nucleus-evoked startle. Kynurenate in the vestibular nucleus had no effect on trigeminal nucleus-evoked startle. These results indicate that trigeminal nucleus stimulation evokes startle largely through glutamate receptors in the PnC, similarly to acoustic startle, but vestibular nucleus-evoked startle is mediated through other pathways, such as the vestibulospinal tract.

Animals↗

Field trial of a new aeration system for enhancing biodegradation in a biopile.

The influence of a new aeration system on the biopile performance was investigated. The purpose was to increase biodegradation efficiency by optimising airflow through the pile. During a 1-month field trial, the performance of a new system using two perforated vertical pipes with wind-driven turbines was compared with that of a standard pile configuration with two horizontal perforated pipes. Both piles were composed of a similar mix of diesel-contaminated soils, woodchips, compost and NPK fertiliser. Hydrocarbons were recovered using solvent extraction, and determined both gravimetrically and by gas chromatography. Total heterotrophs, pH and moisture content were also assessed. Air pressure measurements were made to compare the efficiency of suction in the pipes. Results at the end of the experiment showed that there was no significant difference between the two piles in the total amount of hydrocarbon biodegradation. The normalised degradation rate was, however, considerably higher in the new system than in the standard one, suggesting that the vertical venting method may have improved the efficiency of the biological reactions in the pile. The pressure measurements showed a significant improvement in the suction produced by the new aeration system. However, many factors other than the airflow (oxygen supply) may influence and limit the biodegradation rates, including moisture content, age of contaminants and the climatic conditions. Additional experiments and modelling need to be carried out to explore further the new aeration method and to develop criteria and guidelines for engineering design of optimal aeration schemes in order to achieve maximum biodegradation in biopiles.

Air Movements↗

Degradation of carbazole in the presence of non-aqueous phase liquids by Pseudomonas sp.

Biodegradation of carbazole was enhanced by the presence of a non-aqueous phase liquid (logKo/w > or = 3.1) at phase ratio of 1:1 (organic/aqueous). In a cyclohexane/aqueous phase system, the maximum specific degradation rate (3.34 mg carbazole min(-1) g dry cell(-1)) was at an organic/aqueous ratio of 1:1. Pseudomonas sp. XLDN4-9 degraded 47% (w/w) of 1 g carbazole l(-1) in cyclohexane phase directly within 1 h.

Biodegradation, Environmental↗

Two epithelial cell invasion-related loci of the oral pathogen Actinobacillus actinomycetemcomitans.

Two invasion-related loci, apiA and the two-gene operon apiBC, were isolated from the oral pathogen Actinobacillus actinomycetemcomitans UT32. apiA encodes a 32.5 kDa protein that migrates on SDS-PAGE as a 101 kDa protein as detected by Western blot analysis or silver staining of an outer membrane-enriched fraction of Escherichia coli transformants. E. coli expressing ApiA have a different phenotype than the host vector, in broth and on solid media, and a colony morphology that resembles that of fresh A. actinomycetemcomitans isolates. These E. coli transformants bound to chicken collagen type II, human collagen type II, III, V and fibronectin. apiB and apiC encode proteins of 130.1 and 70.6 kDa, respectively. ApiBC conferred on E. coli a slightly enhanced ability to bind to collagen type III. ApiA- and ApiB-deficient mutants were constructed in A. actinomycetemcomitans. The ApiB-mutant had 4-fold diminished invasion of KB cells; the ApiA-mutant had increased invasion. Both loci were found in all A. actinomycetemcomitans strains, although polymorphism was detected only for apiBC. The deduced sequences of these invasion-related proteins are homologous to members of the YadA adhesin/invasin family.

Adhesins, Bacterial↗

Evaluation of soil gas sampling and analysis techniques at a former petrochemical plant site.

Methods for soil gas sampling and analysis are evaluated as part of a research study on soil vapour intrusion into buildings, conducted at a former petro-chemical plant site ("Chatterton site"). The evaluation process was designed to provide information on reliability and selection of appropriate methods for soil gas sampling and analysis, and was based on a literature review of data and methods, and experiments completed as part of the research study. The broader context of this work is that soil gas characterization is increasingly being used for input into risk assessment of contaminated sites, particularly when evaluating the potential intrusion of soil vapour into buildings. There are only a limited number of research studies and protocols addressing soil gas sampling and analysis. There is significant variability in soil gas probe design and sample collection and analysis methods used by practitioners. The experimental studies conducted to evaluate soil gas methods address the permeation or leakage of gases from Tedlar bags, time-dependent sorption of volatile organic compound (VOC)-vapours onto probe surfaces and sampling devices, and analytical and quality control issues for light gas and VOC analyses. Through this work, common techniques for soil gas collection and analysis are described together with implications for data quality arising from the different methods used. Some of the potential pitfalls that can affect soil gas testing are identified, and recommendations and guidance for improved protocols are provided.

Biodegradation, Environmental↗

Effect of postmolding heat treatment on in vitro properties of a polyanhydride implant containing gentamicin sulfate.

A polyanhydride implant containing gentamicin sulfate was fabricated using a laboratory-scale injection-molding machine. After injection molding, the implants were subject to heat treatment at 60 degrees C for various time periods with or without nitrogen protection. The impact of this heat treatment on the in vitro properties of the implants including copolymer molecular weights, mechanical properties, and in vitro drug-release profiles was investigated. This heat treatment caused a drastic drop in the molecular weight of the copolymer. Heating without nitrogen protection resulted in the hardening of the implant, but heating in the presence of nitrogen rendered the implant less rigid. It was also found that a faster in vitro drug release profile was shown by implants heated without nitrogen protection and a pronounced slowing down in drug release was exhibited by implants heated with nitrogen protection.

Anti-Bacterial Agents↗

Are inequalities in height narrowing? Comparing effects of social class on height in two generations.

OBJECTIVE: To determine whether social inequalities in height change across generations. METHODS: The target population was from the 1958 British birth cohort, all born 3rd-9th March 1958, followed to 1991, and the offspring of one third of this population. Main outcomes were height measured at 7, 11, 16, and 33 years (cohort members) and once at 4-18 years (offspring). Multilevel models applied to associations of social class of origin with (a) child-to-adult growth trajectory (cohort members), (b) height (offspring), and (c) generational height increment. RESULTS: Height inequalities were observed among cohort members, with differences >2.0 cm at all ages between classes I and II, and IV and V. By adulthood, the difference in mean height had declined significantly in boys and slightly in girls. A secular trend was seen between the two generations. While male offspring had a similar mean height to their fathers in classes I and II, boys in classes IV and V gained 2.1 cm (p<0.001). Height gains of female offspring were evident in all classes, with a greater gain in classes IV and V (non-significant). The social class effect on height was weaker among offspring, with a difference between classes I and II, and IV and V of less than 1 cm. CONCLUSIONS: Social inequalities in height observed among the cohort weakened substantially in the next generation due to a greater height gain among offspring from manual classes. Inequalities in childhood height have narrowed between the two generations in this study.

Adolescent↗

Localization of Porphyromonas gingivalis-carrying fimbriae in situ in human periodontal pockets.

Fimbriae, which are involved in adherence, constitute an important pathogenic factor of Porphyromonas gingivalis. In vivo, however, the distribution of P. gingivalis-carrying fimbriae is unknown. The localization of P. gingivalis-carrying fimbriae was examined in situ. From 19 patients with severe periodontitis and P. gingivalis, we obtained 20 teeth with periodontal tissue attached, with and without immunolocalized fimbriae. Eleven teeth were subjected to light microscopy, 9 to electron microscopy. In 6 of the 11 samples examined, we detected positive reactions with an anti-P. gingivalis-fimbriae serum, located in the cementum-attached plaque area in the deep pocket zones. In the so-called 'plaque-free zones', P. gingivalis-carrying fimbriae were immunocytochemically observed to reside in contact with the dental cuticle in 6 of the 9 samples examined. These findings suggest that P. gingivalis-carrying fimbriae are strongly related to adherence to the root surface at the bottoms of human periodontal pockets.

Aged↗

Transgenic mice overexpressing both amyloid beta-protein and perlecan in pancreatic acinar cells.

Heparan sulfate proteoglycans such as perlecan are thought to facilitate amyloid fibril formation. Tg3695 mice overexpress perlecan core protein in many tissues including the brain and pancreas. Tg13592 mice overexpress the signal plus 99-amino acid carboxyl terminal sequences (C99) of amyloid beta-protein precursor in multiple tissues and develop amyloid deposits in the pancreas. To investigate a role of perlecan in beta-amyloidosis, we established doubly transgenic mice by crossing the two lines of transgenic mice. The expression levels of the two transgenes remained unchanged in the brain and pancreas and the doubly transgenic mice did not develop amyloid deposits in the brain up to 19-months of age. Amyloid load detected by thioflavine S in the pancreas of the doubly transgenic mice was not significantly different from that in the transgenic littermates expressing only C99. Amyloid load in the pancreas increased during aging. We found a positive correlation between the Abeta-immunoreactive (non-fibrillar and fibrillar) and thioflavine S-positive (fibrillar) Abeta deposits in the single (C99) but not doubly transgenic mice. Our results suggest that perlecan does not independently influence amyloid formation in the pancreas of the transgenic mice and that there may be other factors that may modulate amyloid formation together with perlecan.

Aging↗

Evaluation of recombination repair pathways in thermal radiosensitization.

Thermal radiosensitization has been shown to cause inhibition of repair of sublethal and potentially lethal damage and DNA DSBs. In this study we assessed thermal radiosensitization in mutants deficient in homologous recombinational (HR) repair and nonhomologous end joining repair (NHEJ). Using cells of the mouse wild-type embryo fibroblast cell line MEF and its Ku80(-/-) derivative that is deficient in NHEJ, we showed that thermal radiosensitization is the same in both cell lines. Further studies with cells of the wild-type CHO-AA8 cell line and its derivative IRS(ISF), which is deficient in HR, also showed comparable thermal radiosensitization in both cell lines. Further experiments using cells of chicken DT40 cell lines also showed comparable thermal radiosensitization between the wild-type HR mutant Rad54, the NHEJ mutant Ku70, and the double mutant Rad 54-Ku70. These results indicate that the HR and NHEJ pathways may not be targets for thermal radiosensitization.

Animals↗

Differential expression of Kir6.1 and SUR2B mRNAs in the vasculature of various tissues in rats.

ATP-sensitive K(+) channels (K(ATP)) couple the intermediary metabolism to cellular excitability and play an important role in the cardio-protective effect of ischemic preconditioning and the activity-dependent autoregulation of cerebral circulation. Although previous studies using PCR and Northern blot suggest that the vascular isoform may consist of Kir6.1 and SUR2B, their expression and precise distribution in various vasculatures remain unknown. To illustrate their vascular expression, we performed this study using in situ hybridization histochemistry. Antisense riboprobes were synthesized by in vitro transcription and labeled with digoxigenin. Distributions of these mRNAs in the various blood vessels were revealed under a bright-field microscope. The expression of Kir6.1 and SUR2B mRNAs was observed in small and intermediate arteries as well as arterioles in several tissues, including basilar, vertebral, mesenteric, coronary and renal arteries. The transcripts were found in arterial smooth muscles. Also, we observed Kir6.1/SUR2B expression in capillary beds. The Kir6.1 and SUR2B expression pattern showed clear overlap, suggesting that they may form heteromeric K(ATP) channels in these tissues. The Kir6.1 and SUR2B stains were detected in aorta and renal tubular cells although their expression level was extremely low. In contrast, the Kir6.1 and SUR2B mRNAs were not seen in vena cava, other small veins, myocardium and skeletal muscles. With their strong expression in small arteries and capillaries, it is very likely that the Kir6.1 and SUR2B form the vascular isoform of K(ATP) channels in these vasculatures.

ATP-Binding Cassette Transporters↗

Cold induces shifts of voltage dependence in mutant SCN4A, causing hypokalemic periodic paralysis.

BACKGROUND: The authors reported a mutation, P1158S, of the human skeletal muscle sodium channel gene (SCN4A) in a family with cold-induced hypokalemic periodic paralysis (hypoKPP) and myotonia. OBJECTIVE: To identify mechanisms of temperature dependency in this channelopathy. METHODS: Using the amphotericin B perforated patch clamp method, sodium currents were recorded at 22 and 32 degrees C from the wild-type (WT) and P1158S mutant SCN4A expressed in tsA201 cells. Computer simulation was performed, incorporating the gating parameters of the P1158S mutant SCN4A. RESULTS: P1158S mutant SCN4A exhibited hyperpolarizing shifts in voltage dependence of both activation and inactivation curves at a cold temperature and a slower rate of inactivation than the WT. Computer simulation reproduced the abnormal skeletal muscle electrical activities of both paralysis at a low potassium concentration in the cold and myotonia at a normal potassium concentration. CONCLUSIONS: Both paralysis and myotonia are attributable to the biophysical properties of the SCN4A mutation associated with hypoKPP. This is the first report of an SCN4A mutation that exhibits temperature-dependent shifts of voltage dependence in sodium channel gating.

Action Potentials↗

Development of highly regenerable callus lines and biolistic transformation of turf-type common bermudagrass [Cynodon dactylon (L.) Pers.].

Common bermudagrass, Cynodon dactylon, is a widely used warm-season turf and forage species in the temperate and tropical regions of the world. Improvement of bermudagrass via biotechnology depends on improved tissue culture responses, especially in plant regeneration, and a successful scheme to introduce useful transgenes. When the concentration of 6-benzylaminopurine was adjusted in the culture medium, yellowish, compact calluses were observed from young inflorescence tissue culture of var. J1224. Nine long-term, highly regenerable callus lines (including a suspension-cultured line) were subsequently established, of which six were used for biolistic transformation. Five independent transgenic events, with four producing green plants, were obtained following hygromycin B selection from one callus line. Three transgenic events displayed resistance to the herbicide glufosinate, and one of these showed beta-glucuronidase activity since the co-transformation vector used in the experiments contained both the gusA and bar genes.

Biolistics↗

Identification of bacteria recovered from clinical specimens by 16S rRNA gene sequencing.

The sequence of the 16S rRNA gene has been used extensively for phylogenetic classification, identification, and genotypic typing of bacteria. Identification of bacterial isolates by 16S rRNA gene sequencing, though generally performed in reference laboratories, has been recently introduced for routine use in clinical laboratories to identify isolates that cannot be identified by conventional methods. Described in this report is the use of 16S rRNA gene sequencing to identify uncommon bacteria, or bacteria with unusual phenotypic properties, with four brief case presentations to illustrate its clinical application. The feasibility, usefulness and limitations of performing this approach in the clinical laboratory are also discussed.

Bacteria↗