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Biomedical subjects

C E Ellis

Publications and source records attributed to C E Ellis.

At least 19 recordsLinked to original sources
PubMed2006

Protective immune responses induced by different recombinant vaccine regimes to Rift Valley fever.

The glycoprotein (GP) and nucleocapsid (NC) genes of Rift Valley fever virus (RVFV) were expressed in different expression systems and were evaluated for their ability to protect mice from virulent challenge using a prime-boost regime. Mice vaccinated with a lumpy skin disease virus-vectored recombinant vaccine (rLSDV-RVFV) expressing the two RVFV glycoproteins (G1 and G2) developed neutralising antibodies and were fully protected when challenged, as were those vaccinated with a crude extract of truncated G2 glycoprotein (tG2). By contrast mice vaccinated with a DNA vaccine expressing G1 and G2 did not sero-convert with only 20% of them surviving challenge. Mice vaccinated with the DNA vaccine and boosted with rLSDV-RVFV also failed to sero-convert but 40% survived challenge. Surprisingly, although none of the mice immunised with the purified NC protein sero-converted, 60% of them survived virulent challenge. The rLSDV-RVFV construct was then further evaluated in sheep for its dual protective abilities against RVFV and sheeppox virus (SPV). Vaccinated sheep sero-converted for both viruses and were protected against RVFV challenge, however, neither the immunised or negative control animals showed any significant reactions to the virulent SPV challenge.

Animals↗
PubMed2000

alpha-synuclein is phosphorylated by members of the Src family of protein-tyrosine kinases.

alpha-Synuclein (alpha-Syn) is implicated in the pathogenesis of Parkinson's Disease, genetically through missense mutations linked to early onset disease and pathologically through its presence in Lewy bodies. alpha-Syn is phosphorylated on serine residues; however, tyrosine phosphorylation of alpha-Syn has not been established (, ). A comparison of the protein sequence between Synuclein family members revealed that all four tyrosine residues of alpha-Syn are conserved in all orthologs and beta-Syn paralogs described to date, suggesting that these residues may be of functional importance (). For this reason, experiments were performed to determine whether alpha-Syn could be phosphorylated on tyrosine residue(s) in human cells. Indeed, alpha-Syn is phosphorylated within 2 min of pervanadate treatment in alpha-Syn-transfected cells. Tyrosine phosphorylation occurs primarily on tyrosine 125 and was inhibited by PP2, a selective inhibitor of Src protein-tyrosine kinase (PTK) family members at concentrations consistent with inhibition of Src function (). Finally, we demonstrate that alpha-Syn can be phosphorylated directly both in cotransfection experiments using c-Src and Fyn expression vectors and in in vitro kinase assays with purified kinases. These data suggest that alpha-Syn can be a target for phosphorylation by the Src family of PTKs.

Amino Acid Sequence↗
PubMed1999

Human urotensin-II is a potent vasoconstrictor and agonist for the orphan receptor GPR14.

Urotensin-II (U-II) is a vasoactive 'somatostatin-like' cyclic peptide which was originally isolated from fish spinal cords, and which has recently been cloned from man. Here we describe the identification of an orphan human G-protein-coupled receptor homologous to rat GPR14 and expressed predominantly in cardiovascular tissue, which functions as a U-II receptor. Goby and human U-II bind to recombinant human GPR14 with high affinity, and the binding is functionally coupled to calcium mobilization. Human U-II is found within both vascular and cardiac tissue (including coronary atheroma) and effectively constricts isolated arteries from non-human primates. The potency of vasoconstriction of U-II is an order of magnitude greater than that of endothelin-1, making human U-II the most potent mammalian vasoconstrictor identified so far. In vivo, human U-II markedly increases total peripheral resistance in anaesthetized non-human primates, a response associated with profound cardiac contractile dysfunction. Furthermore, as U-II immunoreactivity is also found within central nervous system and endocrine tissues, it may have additional activities.

Amino Acid Sequence↗
PubMed1999

Characterization of two human beta1-adrenergic receptor transcripts: cloning and alterations in the failing heart.

Two human beta1-adrenergic receptor cDNAs, approximately 2.75 kb and approximately 3 kb in length, were isolated from a placenta phage library. Both transcripts are collinear with the previously isolated genomic sequence. Additionally, all clones begin between -274 and -236 bp relative to the translational start site, which is consistent with the previously identified transcriptional start site at -263. Furthermore, the 2.75 and 3 kb transcripts utilize conserved polyadenylation consensus sites at +2469 and +2751, respectively. Thus, this is the first report of the identification of full-length human beta1-AR cDNA clones. Both transcripts are expressed in placenta, heart, cerebral cortex, and lung with the 3 kb transcript more highly expressed than the 2.75 kb transcript in all tissues. RNase protection analysis utilizing left ventricular heart RNA isolated from patients with idiopathic dilated cardiomyopathy demonstrates 2.1-fold and 2.7-fold decreases of the 2. 75 and 3 kb transcripts, respectively, as compared with nonfailing controls. Thus, in heart failure patients the 3 kb transcript decreases to a significantly greater extent than the 2.75 kb transcript. This preferential reduction may be the result of differences in mRNA stability mediated by putative AU-rich elements specific to the 3'-untranslated region of the larger transcript.

Base Sequence↗
PubMed1999

Characterization of recombinant human orexin receptor pharmacology in a Chinese hamster ovary cell-line using FLIPR.

The cellular mechanisms underlying the physiological effects of the orexins are poorly understood. Therefore, the pharmacology of the recombinant human orexin receptors was studied using FLIPR. Intracellular calcium ([Ca2+]i) was monitored in Chinese hamster ovary (CHO) cells stably expressing orexin-1 (OX1) or orexin-2 (OX2) receptors using Fluo-3AM. Orexin-A and orexin-B increased [Ca2+]i in a concentration dependent manner in CHO-OX1 (pEC50=8.03+/-0.08 and 7. 30+/-0.08 respectively, n=5) and CHO-OX2 (pEC50=8.18+/-0.10 and 8. 43+/-0.09 respectively, n=5) cells. This response was typified as a rapid peak in [Ca2+]i (maximal at 6 - 8 s), followed by a gradually declining secondary phase. Thapsigargin (3 microM) or U73122 (3 microM) abolished the response. In calcium-free conditions the peak response was unaffected but the secondary phase was shortened, returning to basal values within 90 s. Calcium (1.5 mM) replacement restored the secondary phase. In conclusion, orexins cause a phospholipase C-mediated release of calcium from intracellular stores, with subsequent calcium influx.

Aniline Compounds↗
PubMed1999

In vitro evaluation methods for Clostridium botulinum type C and D vaccines.

Reagents were prepared for use in ELISAs to determine the concentration of the antigenic components of Clostridium botulinum type C and D. The results obtained were compared with the L+dose assay and a good correlation was found between the two assays for measurement of the C and D neurotoxin concentration. These ELISAs were also used to determine the concentration of the neurotoxins in toxoid form. The relationship between the C neurotoxin dose, in toxoid form, and the immune response in guinea pigs could be deduced from the data obtained. The relationship for the D neurotoxin was not that clear, as the same concentration of the antigen resulted in variable potency values. However, these ELISAs can be used to formulate the concentration of the C and D components in the final bivalent vaccine. Replacement of the preliminary potency assay on the monovalent components after production with the in vitro assays will shorten the total production time of the vaccine by about 60 days. The economical and ethical implications are the reduction in the use of animals to evaluate the vaccine.

Animals↗
PubMed1999

The production and evaluation of Pasteurella haemolytica leukotoxin in the supernatant of submerged cultures in fermenters.

The optimal production of P. haemolytica leukotoxin in the culture supernatant of a fluid medium is dependent on a number of factors. The leukotoxin has to be produced by using a strain that is known for its ability to produce high quantities of leukotoxin, inoculated into the most suitable type of medium at the correct culture density containing the necessary supplements and harvested after a certain growth period. The volume in which it is produced may also have an influence. Two different procedures are described to produce the leukotoxin in 5 to 15-l quantities in RPMI 1640 medium. The first method used to produce leukotoxin is one that has been repeatedly described since the presence of the leukotoxin was first established in 1978. Using this method seven batches of leukotoxin were produced in litre quantities with leukotoxin activity ranging from 23-67 u/ml. The seed culture inoculum is prepared in brain heart infusion broth, which is centrifuged before the organisms are inoculated into RPMI 1640 medium containing 3.5% foetal calf serum and incubated for only 1 h in a fermenter, after, which the leukotoxin is harvested. An improved alternative method was devised which yielded higher levels of leukotoxin activity by utilising the ability of the P. haemolytica organisms to grow and produce leukotoxin during the logarithmic growth phase in a fermenter. A seed culture harvested in the log phase was prepared in brain heart infusion broth by means of a series of cultures and inoculated into RPMI 1640 containing 3.5% foetal calf serum. Three hours of active growth were allowed during which the leukotoxin was measured by its biological activity and an ELISA assay, and the increase in cell mass by means of the optical density every 30 min. The average leukotoxin biological activity measured 260 u/ml and by means of the ELISA test the leukotoxin concentration measured 315 u/l which is a substantial increase in leukotoxin production. In comparison the average optical density only measured 0.469 at 650 nm. Previous findings were substantiated that the highest cell density was not reflected in the highest leukotoxin activity. It is possible to induce high levels of leukotoxin secretion in submerged cultures with RPMI 1640 medium containing foetal calf serum in the controlled environment of a fermenter in large enough quantities for use as a vaccine by the improved preparation of the seed culture inoculum.

Absorption↗
PubMed1998

Preliminary analysis of the transcriptional regulation of the human beta 1-adrenergic receptor gene.

One open reading frame of a 13 kb genomic clone of the human beta 1-adrenergic receptor, which lacks introns, encodes the previously isolated cDNA. Transcript(s) between 4.7 and 5.1 kb are detected in total RNA, whereas a approximately 3 kb transcript is detected only in polyadenylated RNA. The poly (A+) transcript is most highly expressed in the pancreas, liver, heart, kidney, thalamus, adrenal, and salivary glands. Primer extension and ribonuclease protection analyses suggest that the major transcriptional start site is located at -263. Transient expression of luciferase reporter gene constructs indicates that the region from -444 to -360 possesses the primary promoter, consistent with the transcriptional start site at -263. Negative transcriptional regulatory elements are located from -3118 to -2730 and -2730 to -2241, while a positive element is located between -2241 and -1790. The present study suggests that, despite similarities, the expression and transcriptional regulation of the human gene are distinct from those of the genes of other species.

Animals↗
PubMed1998

Functional characterisation of the human cloned 5-HT7 receptor (long form); antagonist profile of SB-258719.

1. The functional profile of the long form of the human cloned 5-HT7 receptor (designated h5-HT7(a)) was investigated using a number of 5-HT receptor agonists and antagonists and compared with its binding profile. Receptor function was measured using adenylyl cyclase activity in washed membranes from HEK293 cells stably expressing the recombinant h5-HT7(a) receptor. 2. The receptor binding profile, determined by competition with [3H]-5-CT, was consistent with that previously reported for the h5-HT7(a) receptor. The selective 5-HT7 receptor antagonist SB-258719 ((R)-3,N-Dimethyl-N-[1-methyl-3-(4-methylpiperidin-1-yl)propyl]ben zene sulfonamide) displayed high affinity (pKi 7.5) for the receptor. 3. In the adenylyl cyclase functional assay, 5-CT and 8-OH-DPAT were both full agonists compared to 5-HT and the rank order of potency for agonists (5-CT > 5-HT > 8-OH-DPAT) was the same in functional and binding studies. 4. Risperidone, methiothepin, mesulergine, clozapine, olanzapine, ketanserin and SB-258719 antagonised surmountably 5-CT-stimulated adenylyl cyclase activity. Schild analysis of the antagonism by SB-258719 gave a pA2 of 7.2+/-0.2 and slope not significantly different from 1, consistent with competitive antagonism. 5. The same antagonists also inhibited basal adenylyl cyclase activity with a rank order of potency in agreement with those for antagonist potency and binding affinity. Both SB-258719 and mesulergine displayed apparent partial inverse agonist profiles compared to the other antagonists tested. These inhibitory effects of antagonists appear to be 5-HT7 receptor-mediated and to reflect inverse agonism. 6. It is concluded that in this expression system, the h5-HT7(a) receptor shows the expected binding and functional profile and displays constitutive activity, revealing inverse agonist activity for a range of antagonists.

Adenylyl Cyclases↗
PubMed1996

On the mental representation of conditional sentences.

Four experiments are reported which attempt to externalize subjects' mental representation of conditional sentences, using novel research methods. In Experiment 1, subjects were shown arrays of coloured shapes and asked to rate the degree to which they appeared to be true of conditional statements such as "If the figure is green then it is a triangle". The arrays contained different distributions of the four logically possible cases in which the antecedent or consequent is true or false: TT, TF, FT, and FF. For example, a blue triangle would be FT for the conditional quoted above. In Experiments 2 to 4, subjects were able to construct their own arrays to make conditional either true or false with any distribution of the four cases they wished to choose. The presence and absence of negative components was varied, as was the form of the conditional, being either "if then" as above or "only if": "The figure is green only if it is a triangle". The first findings was that subjects represent conditional in fuzzy way: conditional that include some counter-example TF cases (Experiment 1) may be rated as true, and such cases are often included when subjects construct an array to make the rule true (Experiments 2 to 4). Other findings included a strong tendency to include psychologically irrelevant FT and FF cases in constructed arrays, presumably to show that conditional statements only apply some of the time. A tendency to construct cases in line with the "matching bias" reported on analogous tasks in the literature was found, but only in Experiment 4, where the number of symbols available to construct each case was controlled. The findings are discussed in relation to the major contemporary theories of conditional reasoning based upon inference rules and mental models, neither of which can account for all the results.

Adult↗
PubMed1981

A survey of post mortem findings in 480 horses 1958 to 1980: (1) causes of death.

The causes of death recorded in 480 consecutive post mortem examinations of horses performed at the department of pathology, Veterinary Field Station, University of Liverpool, between February 1958 and February 1980, are reported. The alimentary, locomotor and nervous systems were those most frequently diseased. The most common specific entities were those associated with grass sickness, fractures and endoparasitism.

Animals↗
PubMed1981

A survey of post mortem findings in 480 horses 1958 to 1980: (2) disease processes not directly related to the cause of death.

Disease processes not directly related to the cause of death recorded in 480 consecutive post mortem examinations of horses performed at the department of pathology, Veterinary Field Station, University of Liverpool, between February 1958 and February 1980 are reported. The alimentary, cardiovascular, respiratory and locomotor systems were those most frequently diseased. The most common specific entities were those associated with endoparasitism and its associated vascular lesions, pneumonia and fractures.

Animals↗
PubMed1980

Pasteurella multocida infection of poultry farm rats.

Thirty-four rat carcases from 11 poultry farms were examined for the presence of Pasteurella multocida; 14 out of 34 (41 per cent) proved positive after mouse inoculation, compared with five out of 34 (14 per cent) using media alone. No salmonellae were recovered from 27 rat carcases using enrichment media. Poultry pasteurellosis was present on two farms with infected rats and the same serotype was present in rats and poultry in those cases.

Animals↗
PubMed1980

Virulence and morphology of Pasteurella multocida of avian origin.

The virulence of 100, 54 and nine avian isolates of Pasteurella multocida for mice, chickens and turkey poults respectively was assessed and rated on a scale from 0 to 5. Cellular and colonial morphology of the bacteria was examined after mouse passage. There was evidence of a considerable range in the virulence of the isolates for the test species, but no correlation between cellular or colonial morphology and virulence was detected. The results obtained are discussed in relation to general epidemiological considerations.

Animals↗