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P Fu

Publications and source records attributed to P Fu.

At least 73 records · Page 4Linked to original sources

Erythropoietin gene expression in fetal and adult sheep kidney.

The exact location of the cells which express erythropoietin (Epo) in the kidney is still controversial, with conflicting reports suggesting that both peritubular interstitial cells or proximal tubular epithelial cells are possible sites of Epo expression. In the present study we have examined the location of Epo-expressing cells in fetal and adult sheep kidneys after Epo expression was stimulated by anaemia. In situ hybridization histochemistry was performed using synthetic oligonucleotide probes complementary to part of the sheep Epo cDNA sequence or a riboprobe (cRNA) for ovine Epo of 520 bases. Epo expression was confined to peritubular cells of the kidney cortex, in general in the area close to the cortico-medullary junction. In some severely anaemic adult sheep kidneys, Epo-expressing cells were also found in the outer cortex. In addition we located Epo-expressing cells in the kidneys not only of anaemic fetuses (89-140 d of gestation, term = 150d) but also in kidneys from normal fetuses 60-110d of gestation. Again, Epo expression was seen only in peritubular cells of the kidney cortex. These findings confirm that the kidney is an important site of Epo production, in the sheep, from at least 0.4 gestation, but also show that there is no ontogenic change in the cellular site of production within the kidney.

Animals↗

A study on the structure of human optic nerve lamina cribrosa.

PURPOSE: To determine the mechanism of nerve fiber damage in glaucoma by studying the structure of human optic nerve lamina cribrosa (LC) in different regions. METHODS: 15 human eyes of 10 cases were studied. The specimens were prepared for scanning electron microscopy, and numbers and areas of pores in LC were measured by electron image analysis system. Draw a frequency distribution map with each curve represents the tendency of pores distribution in a particular part. The proportion of the connective tissue in respective quadrant can also be calculated. The specimens were also prepared for histological examination. RESULTS: There are many pores of various magnitude and shapes on the surface of LC. There are significantly more large pores (> or = 3000 micron2) in the superior and inferior than those in the nasal and temporal quadrants, especially in the peripheral regions. In terms of area, the percentages of connective tissue in the nasal and temporal quadrants are the highest. Collageous fibers, various in diameter, are arranged in bundles and tangentially around each pore. CONCLUSION: In normal persons, the percentage of large pores in the superior and inferior peripheral parts is the highest, the density of the connective tissue is the lowest. So, the force received by unit area of the superior and inferior parts is bigger than that of the nasal and temporal sides, therefore, it is susceptible to the impact of high intraocular pressure at the early stage and causing corresponding visual defect. Our study may suggest the mechanism of optic nerve damage of glaucoma.

Adult↗

Methods and strategies available for the process control and optimization of monoclonal antibody production.

The objective of this paper is to explore the range of methods and strategies available for the process control and optimization of monoclonal antibody production by hybridoma cell culture. Emphasis will be placed on the choice of the level of complexity incorporated into the process control and optimisation procedure. It will be shown that the behaviour of hybridomas in culture is influenced by sophisticated cellular metabolic activities and various interactive environmental factors and that the understanding and modelling of the way hybridomas grow in the bioreactor should enable optimisation of bioreactor operating conditions to achieve maximum monoclonal antibody formation. However, due to the lack of on-line instrumentation of important biological variables and the incomplete knowledge of hybridoma cultivation process, there exist many limitations and challenges to the advent of applications of process control and optimisation in this field. To solve the problem, introduction of industrially practical biological measurements and development of new control concepts are inevitable. At the end of this paper, we shall discuss possible schemes for the control of the physiological state of cells in order that balanced cell growth and maximum monoclonal antibody synthesis may be achieved.

Animals↗

Characterization of two relaxin genes in the chimpanzee.

Relaxin is a peptide hormone which has a variety of physiological effects on tissues of the reproductive tract as well as other organs such as the heart and brain. Whereas all non-primates so far examined have only a single relaxin gene, humans have two genes (H1 and H2, or gene 1 and gene 2). H2 relaxin is synthesized in the corpus luteum during pregnancy and is also found in the placenta and prostate, whereas expression of H1 has been very difficult to detect. We have begun a study of relaxin genes in the chimpanzee to assess whether this species may provide a suitable model in which to examine the roles of gene 1 relaxin. We find that the chimpanzee has two relaxin genes, one of which is very similar to H2. The second gene has an gene 1 type A chain but the B chain is of the gene 2 type, possibly due to a gene conversion event. The authentic chimpanzee gene 2 (Ch2) is expressed in the corpus luteum of pregnancy and in the placenta. Ch1 is not expressed in the placenta, but the mRNA can be detected by polymerase chain reaction in the corpus luteum.

Amino Acid Sequence↗

[A study on the structure of human lamina cribrosa sclerae].

The lamina cribrosa of 15 normal adult human eyes were observed by scanning electron microscopy and their images were analyzed by a computerized system to study the distribution of pores and the proportion of connective tissue in different regions of the lamina cribrosa. The results revealed that there are significantly more large pores (> or = 3,000 microns2) in the superior and inferior than those in the nasal and temporal quadrants, especially in the peripheral regions, however, in terms of area the percentages of connective tissue in the nasal and temporal quadrants are the highest. In addition, the histological study of the lamina cribrosa disclosed that the collagenous fibers, various in calibre, are arranged in bundles and tangentially around each pore. Since the nerve fiber damage in glaucoma initially occurs at the superior and inferior parts of the optic nerve, the above anatomical characteristics possibly can explain the cause of the visual field defects in early glaucoma.

Adult↗

The sheep erythropoietin gene: molecular cloning and effect of hemorrhage on plasma erythropoietin and renal/liver messenger RNA in adult sheep.

Erythropoietin (Epo) production was studied in adult sheep. Nine ewes, body weight (BW) 39 +/- 1.3 kg, were hemorrhaged a volume of blood equivalent to 1.6% BW, and sampled at 0, 2, 4, 6, 24 h. Oxygen content (O2 CT) decreased by 2.7 +/- 0.6 ml/dl at 2 h. Plasma immunoreactive (IR) Epo was only significantly increased at 24 h, from 18.5 +/- 3.5 to 40 +/- 10.7 mU/ml (mean +/- SEM). A further 5 ewes were bled extensively (2793 +/- 82 ml) over 54 h, and killed for Epo mRNA determination. The O2 CT decreased from 12.3 +/- 1.6 to 4.1 +/- 0.6 ml/dl, and plasma Epo increased from 15 +/- 4 to 1675 +/- 287 mU/ml. The sequence of ovine Epo cDNA was derived from the kidney RNA of a severely bled sheep using reverse transcription/polymerase chain reaction (RT/PCR), and from an ovine Epo genomic clone. The cDNA encodes a peptide of 194 amino acids, including a 27 amino acid signal peptide. The deduced amino acid sequence of sheep Epo shows 82%, 78% and 80% homology with mature Epos of human, mouse and monkey, respectively. The gene structure resembles closely those of human and mouse, with 5 exons and 4 introns. The expression of the ovine Epo gene in tissues from normal and hemorrhaged sheep was analysed by a competitive RT/PCR method. Epo mRNA was difficult to detect in liver from normal sheep, but was detectable at 0.01-0.04 amol/microgram total RNA in kidney from normal sheep. In the kidneys of severely bled sheep, the Epo mRNA levels (per micrograms total RNA) increased 400-1500-fold compared to that of normal kidneys, and were approximately 60-fold greater than those in the livers of the hemorrhaged sheep.

Amino Acid Sequence↗

Identification of murine homologues of the Drosophila son of sevenless gene: potential activators of ras.

Several findings suggest that signals from tyrosine kinases are transduced, at least in part, through ras proteins. These findings include (i) blockage of the transforming activity of constitutively active tyrosine kinases by inhibiting ras function and (ii) genetic screens in Caenorhabditis elegans and in Drosophila that identified ras genes as downstream effectors of tyrosine kinases. The recently isolated Drosophila gene Son of sevenless (Sos) is postulated to act as a positive regulatory link between tyrosine kinase and ras proteins by catalyzing exchange of GDP for GTP on ras protein. Such exchange proteins have been reported in extracts of mammalian cells but have not been previously characterized at a molecular level. As Sos appears to function in this role in Drosophila, we sought to isolate a vertebrate counterpart(s). We have characterized two widely expressed murine genes with a high degree of homology to Sos. Hybridization with human DNA and RNA indicates a high degree of conservation of these genes in other vertebrates.

Amino Acid Sequence↗

Simulation of an iterative learning control system for fed-batch cell culture processes.

This paper describes an iterative learning control scheme for fed-batch operation where repetitive trajectory tracking tasks are required. The proposed learning strategy is model-independent, and it takes advantage of the repetitive feature of system operations with a certain degree of intelligence and requires only small size of dynamic database for the learning process. The convergence of the learning process is proven. An example of simultaneously tracking two predefined trajectories by iterative learning control with two control inputs is given to illustrate the methodology. Satisfactory performance of the learning system can be observed from the simulation results.

Animals↗

Ethanol uncouples dentate granule neurons by increasing junctional resistance: a multineuronal system model approach.

The effects of an acute intoxicating concentration of ethanol (50 mM) on the electrotonic membrane properties of hippocampal dentate granule neurons were studied using a system model incorporating electrotonic coupling between neurons. Uncoupling of cells by other alcohols has been shown in several tissues. The system model allows a quantitative estimation of the changes in coupling and other neuronal electrotonic properties. The input impedance of a neuron was measured from the voltage decay of a short hyperpolarizing current pulse. An analytic expression of the input impedance has been written incorporating somatic, dendritic, and electrical coupling parameters. Using this particular current stimulation, the modelling results showed that ethanol selectively increased the junctional resistance by more than 2.5 times, hence uncoupling the neurons. A 30% increase in the final time-constant, tau 0, was also obtained from the voltage transient. Other parameters were not significantly affected. A neuronal model without electrotonic coupling to other neurons gave rise to physiologically impossible values for the membrane resistance and capacitance. With resistive and capacitive coupling in the model, uncoupling did not occur with ethanol. It is concluded that ethanol uncouples neurons by increasing the effective gap junctional resistance in dentate granule neurons.

Animals↗

The sequence and tissue expression of ovine renin.

The primary structure of the sheep renin precursor has been determined from its cDNA sequence. A library of cDNA clones was constructed from adrenalectomized sheep kidney poly(A)+ RNA and screened for sheep renin sequences with a cloned mouse renin cDNA probe. Of the 300,000 clones generated, 24 were hybridization positive and the nucleotide sequences of two of the longest clones were determined. These clones coded for the mature sheep renin protein and the 3'-untranslated sequence but did not extend to the amino-terminal region of preprorenin. Clones corresponding to the 5' region of renin mRNA were generated by the polymerase chain reaction and their nucleotide sequences determined. The sheep renin precursor consists of 400 amino acids with a putative leader sequence of 14 amino acids and a putative 45 or 53 amino acid prosegment. The mature sheep renin protein has a 73% sequence identity with human renin. Northern analysis demonstrated the presence of renin mRNA in the kidney but not in other tissues in the sheep. While sodium depletion of sheep caused a rise in renin mRNA in the kidney, adrenalectomy also led to a large increase in renal renin mRNA. Southern analysis of genomic DNA suggests that there is only one gene coding for renin in the sheep.

Amino Acid Sequence↗

Human secreted carbonic anhydrase: cDNA cloning, nucleotide sequence, and hybridization histochemistry.

Complementary DNA clones coding for the human secreted carbonic anhydrase isozyme (CA VI) have been isolated and their nucleotide sequences determined. These clones identify a 1.45-kb mRNA that is present in high levels in parotid submandibular salivary glands but absent in other tissues such as the sublingual gland, kidney, liver, and prostate gland. Hybridization histochemistry of human salivary glands shows mRNA for CA VI located in the acinar cells of these glands. The cDNA clones encode a protein of 308 amino acids that includes a 17 amino acid leader sequence typical of secreted proteins. The mature protein has 291 amino acids compared to 259 or 260 for the cytoplasmic isozymes, with most of the extra amino acids present as a carboxyl terminal extension. In comparison, sheep CA VI has a 45 amino acid extension [Fernley, R. T., Wright, R. D., & Coghlan, J. P. (1988b) Biochemistry 27, 2815]. Overall the human CA VI protein has a sequence identity of 35% with human CA II, while residues involved in the active site of the enzymes have been conserved. The human sheep secreted carbonic anhydrases have a sequence identity of 72%. This includes the two cysteine residues that are known to be involved in an intramolecular disulfide bond in the sheep CA VI. The enzyme is known to be glycosylated and three potential N-glycosylation sites (Asn-X-Thr/Ser) have been identified. Two of these are known to be glycosylated in sheep CA VI. Southern analysis of human DNA indicates that there is only one gene coding for CA VI.

Amino Acid Sequence↗