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T G Cooper

Publications and source records attributed to T G Cooper.

At least 163 records · Page 9Linked to original sources

The allantoin and uracil permease gene sequences of Saccharomyces cerevisiae are nearly identical.

We have determined the structure of the allantoin permease (DAL4) gene of Saccharomyces cerevisiae. The gene putatively encodes a hydrophobic protein with a M(r) of 71,755. It possesses the alternating hydrophobic-hydrophilic regions similar to those found in many other integral membrane proteins. The most striking feature of the allantoin permease component encoded by DAL4 is its striking similarity to the uracil permease component encoded by FUR4. Although data available indicate that these proteins do not share any overlap of function, their predicted protein sequences are 68% identical, 81% similar, and their DNA sequences are 70% identical. The upstream regulatory region of DAL4 contains all of the characterized cis-acting elements previously reported for inducible allantoin pathway genes: six sequences homologous to UASNTR, the element responsible for nitrogen catabolite repression-sensitive activation of allantoin pathway gene expression, and two sequences homologous to the cis-acting element responsible for inducer-responsiveness of the allantoin pathway genes, UIS. The finding of these homologous sequences predicted to exist on the basis of DAL4's expression characteristics, supports and strengthens the suggestion that these elements mediate the functions we have previously ascribed to them.

Amino Acid Sequence↗

Composition of fluids obtained from human epididymal cysts.

The fluid composition of five epididymal spermatocoeles, one epididymal cyst and a hydrocoele was examined. The fluid obtained from the spermatocoeles was a dilute suspension of mainly immotile spermatozoa. The sperm-free fluid contained less protein, phosphate, glucose, triglyceride and cholesterol than serum but more testosterone and chloride than peripheral blood. It contained no epididymal secretion products. Proteins in the fluid differed from those in serum. From the fluid composition these cysts appeared to be continuous with the rete testis, either dilatations of efferent ducts or Haller's superior aberrant duct (vas aberrans of the rete testis). Fluid from an epididymal cyst containing no spermatozoa was mainly of similar composition. In contrast, hydrocoele fluid resembles blood serum.

Adult↗

Appearance and endocytic activity of epithelial cells from human efferent ducts in primary monolayer culture.

The culture of epithelial cells lining human efferent ducts, obtained from prostatic carcinoma patients, is described. Ciliated cells were observed to beat for at least one month on plastic. On previous filters low cuboidal cells characterized the monolayers. Cells comprising monolayers over the filter were 5 to 9 microns in height whereas taller cells were found over the original fragments (14 microns). Some non-ciliated cells contained dark and light vacuoles, others were found to lack them. Both non-ciliated and ciliated cells maintained tight junctional complexes restricting the paracellular movement of horseradish peroxidase. Both types of cultured cells exhibited fluid-phase and adsorptive endocytosis from both apical and basal surfaces. It is reported for the first time that the monolayers form high resistance barriers (150 omega.cm2) that prevent the apical medium from draining to the basal compartment over 24 h.

Cell Polarity↗

Physiologic alterations in cranial blood flow demonstrated by magnetic resonance angiography.

Two-dimensional phase contrast magnetic resonance angiography (MRA) was used to image alterations in cranial blood flow induced by changes in arterial PCO2 in an animal model. MRA was performed on five sheep; 64 acquisitions were obtained in each of three flow encode directions using a 256 x 256 matrix. Sheep were intubated and ventilated with oxygen and 1.5% halothane to prevent any movement. Femoral arterial cannulation was performed to monitor arterial blood gases and pressure. The sheep was secured in a cradle with its head and neck in a 6-inch imaging coil within the 26-cm-clear bore. Images were obtained during separate physiologic states, which were induced by changes in ventilatory parameters. These were normocapnia (PCO2 35-45 mm Hg), hypercapnia (greater than 90-130 mm Hg), and hypercapnia with superimposed hypoxia. Comparisons of images were performed using both a video flashback mode and image subtraction. The authors noted that 1) both venous and arterial flow velocity qualitatively increased during hypercapnia; 2) in addition to change in the caliber of blood vessels, redistribution of blood flow within the cranium could be demonstrated during the PCO2 changes; and 3) blood was directed away from superficial structures and toward the brain during superimposed hypoxia. MRA, previously used to show steady-state cranial flow also can demonstrate flow responses to physiologic stimuli.

Animals↗

Nitrogen catabolite repression of arginase (CAR1) expression in Saccharomyces cerevisiae is derived from regulated inducer exclusion.

Expression of the Saccharomyces cerevisiae arginase (CAR1) gene is regulated by induction and nitrogen catabolite repression (NCR). Arginine was demonstrated to be the native inducer. CAR1 sensitivity to NCR has long been accepted to be accomplished through a negative control mechanism, and cis-acting sites for it have been hypothesized. In search of this negatively acting site, we discovered that CAR1 sensitivity to NCR derives from regulated inducer (arginine) exclusion. The route of catabolic entry of arginine into the cell, the general amino acid permease (GAP1), is sensitive to NCR. However, CAR1 expression in the presence of sufficient intracellular arginine is NCR insensitive.

Arginase↗

Tripartite structure of the Saccharomyces cerevisiae arginase (CAR1) gene inducer-responsive upstream activation sequence.

Arginase (CAR1) gene expression in Saccharomyces cerevisiae is induced by arginine. The 5' regulatory region of CAR1 contains four separable regulatory elements--two inducer-independent upstream activation sequences (UASs) (UASC1 and UASC2), an inducer-dependent UAS (UASI), and an upstream repression sequence (URS1) which negatively regulates CAR1 and many other yeast genes. Here we demonstrate that three homologous DNA sequences originally reported to be present in the inducer-responsive UASI are in fact three exchangeable elements (UASI-A, UASI-B, and UASI-C). Although two of these elements, either the same or different ones, are required for transcriptional activation to occur, all three are required for maximal levels of induction. The elements operate in all orientations relative to one another and to the TATA sequence. All three UASI elements bind protein(s); protein binding does not require arginine or overproduction of any of the putative arginine pathway regulatory proteins. The UASI-protein complex was also observed even when extracts were derived from arg80/argRI or arg81/argRII deletion mutants. Similar sequences situated upstream of ARG5,6 and ARG3 and reported to negatively regulate their expression are able to functionally substitute for the CAR1 UASI elements and mediate reporter gene expression.

Arginase↗

Ty insertions upstream and downstream of native DUR1,2 promoter elements generate different patterns of DUR1,2 expression in Saccharomyces cerevisiae.

Expression of allantoin pathway genes is subject to induction and nitrogen catabolite repression. Two classes of cis-dominant mutations (DUR80 and DUR1,2-Oh) result in overproduction of DUR1,2 mRNA. In DUR80 mutants, DUR1,2 expression remained inducible, nitrogen catabolite repression sensitive, and unresponsive to cell ploidy, i.e., overproduction was superimposed on normal gene regulation. DUR1,2-Oh mutations, in contrast, generated a pattern of DUR1,2 expression similar to that often reported when a Ty element inserts upstream of a gene, the ROAM phenotype. We analyzed four independent DUR80 and DUR1,2-Oh alleles. The DUR1,2-Oh mutation was, as expected, a Ty insertion at -445 3' of the native DUR1,2 upstream activation sequences (UASs). All three DUR80 alleles were also Ty insertions between -644 and -653 immediately 5' of the native DUR1,2 USASs. We suggest that the difference in DUR1,2-Oh and DUR80 phenotypes depends on whether the native cis-acting elements and transcription factors associated with them can operate. If they can, enhancement of normally regulated DUR1,2 expression is observed. This is a novel phenotype for Ty insertions. If the native DUR1,2 cis-acting elements are not present, the case when Ty insertion occurs 3' of them, a ROAM phenotype is generated. Nitrogen-regulated upstream activation sequence (UASNTR)-homologous sequences present in the Ty delta elements rather than cis-acting elements required for Ty transcription are the most likely candidates to serve as the cis-acting elements mediating the DUR80 phenotype.

Base Sequence↗

Characterization of the motility of maturing rat spermatozoa by computer-aided objective measurement.

A computer-aided sperm analysis system was optimized for objective assessment of the movement characteristics of mature and immature rat spermatozoa by testing different settings. Measurements of straight line velocity of individual motile cells were validated by manual tracking with a digitizer. Better agreement between the two methods and better performance in distinguishing between mature and immature spermatozoa was obtained by reducing the tracking rate to increase the time of analysis. However, numbers of motile and immotile cells could not be determined accurately. Manual counting of videotaped images revealed no significant differences in percentage motility of spermatozoa from five epididymal regions. Caput spermatozoa were characterized by low straight-line (VSL) and averaged-path (VAP) velocities and low path straightness (STR), whereas mature cells displayed high VSL, VAP and STR. An increase in curvilinear velocity on maturation was less obvious. Spermatozoa in the proximal corpus epididymidis were heterogeneous in their acquisition of motility maturation and the uniformity of movement pattern achieved in the distal corpus and proximal cauda regions tended to decrease again in the distal cauda epididymidis. Such objective measurements of motility patterns will facilitate studies on the regulation of motility development upon sperm maturation.

Animals↗

Prospects for the future: magnetic resonance fast scanning and flow quantitation.

High-speed scanning techniques are quickly emerging to provide new applications in the MRI domain. These potential applications range from the reduction of patient examination time to the functional imaging of human cognition. Additionally, flow quantitation is rapidly developing into a clinical diagnostic tool available for routine use. This article provides the concepts to acquire a rudimentary understanding of both topics.

Artifacts↗

Internal quality control of semen analysis.

OBJECTIVE: To test a scheme for quality control of semen analysis. DESIGN: The reproducibility of assessment of sperm concentration, motility, and morphology was obtained for the same sample measured by different technicians (between or intertechnician variation) and for different samples assessed by each technician with time (within or intratechnician variation). SETTING: Andrology Laboratory. PATIENTS: Semen samples were analyzed from all patients attending the clinic. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Within technician and between technician coefficients of variation for concentration, motility, and morphology. RESULTS: When 100 sperm were routinely assessed, both intratechnician variation, as assessed from the precision of duplicate measurements, and intertechnician variation revealed hyperbolic curves with increasing variation at low percentages (less than 20) of motile or morphological forms. When these low values were excluded, mean intratechnician variations were 5.0%, 8.4%, and 2.8% for concentration, motility, and morphology, respectively, and mean intertechnician variations were, respectively, 6.1%, 5.6%, and 5.6%. Similar mean intertechnician variation for morphology was obtained for repeated assessment of prestained (7.3%) or presmeared (5.9%) slides. The use of cryopreserved semen to monitor longitudinal changes in the technicians' assessments revealed variations of 8.1% to 12% in concentration and 9.7% to 14% in motility. Computing the monthly means for sperm concentration, motility, and morphology over a 4.5-year period revealed a marked reduction in percentage of normal morphological forms, unrelated to the sperm count or mean age of the men attending the clinic. This was shown to be a result of a shift in the assessment by technicians. CONCLUSIONS: Quality control is necessary and possible in an andrology clinic.

Cryopreservation↗

Sequences of two adjacent genes, one (DAL2) encoding allantoicase and another (DCG1) sensitive to nitrogen-catabolite repression in Saccharomyces cerevisiae.

Reported are the nucleotide sequences of the yeast allantoicase-encoding gene (DAL2) and that of an unknown gene adjacent to it. Expression of the unidentified gene is sensitive to nitrogen catabolite repression (NCR) and regulated by the DAL80 product, a previously documented control element regulating allantoin pathway gene expression. Both genes possess multiple upstream activation sequences (UAS) homologous to the UASNTR element shown to be required for sensitivity to NCR. Also present upstream from DAL2 is a mutant form of the upstream induction sequence required for response of DAL7 to induction. Its occurrence in mutant form is consistent with the poor induction of DAL2 expression observed in vivo.

Allantoin↗

Organization of tubules in the human caput epididymidis and the ultrastructure of their epithelia.

The structure of the human caput epididymidis was examined by gross morphological and light and electron microscopic techniques. There were at least seven types of tubules, each characterized by a different epithelium. These tubules were connected with one another by at least eight types of junctions to form a network. Most of the caput epididymidis was composed of efferent ducts. Within these, five types of tubules, each with a different ciliated epithelium, were found in different regions; and four types of junctions between the efferent ducts and the epididymal tubule were observed. The efferent ducts left the testis, initially as parallel straight tubules containing both ciliated and non-ciliated cells in an epithelium of irregular height. Each efferent duct then coiled tortuously into lobules that folded over one another. These efferent ducts then branched out as thin tubules to join a network of dark tubules which were lined by a regular epithelium containing prominently vacuolated, non-ciliated cells. These tubules anastomosed via common cavities characterized by a ciliated cuboidal epithelium and sometimes joined tubules exhibiting a non-vacuolated ciliated epithelium. The latter, as well as typical efferent ducts, made connection with the epididymis proper in both end-to-end and end-to-side junctions. In the more distal junctions with the epididymis, the efferent ducts joined to a transitional epididymal ductule before joining to the side of the epididymis proper. Post-junctional epithelia in the beginning of the epididymis occasionally contained patches of cells characteristic of efferent ducts. Tall cells with long stereocilia constituted a discontinuous "initial segment"-like region of the epididymis. This is the most detailed study so far of the epithelia and the tubule organization in the caput epididymidis of any species, and most of the results are reported for the first time for the human. Although the pattern of the tubule network resembles that of some domestic species, the rich variety of epithelia has not been appreciated before.

Aged↗

The ureidoglycollate hydrolase (DAL3) gene in Saccharomyces cerevisiae.

The DAL3 gene has been sequenced and found to encode a 195 amino acid protein with a molecular weight of 21,727. The four carboxy-terminal amino acids of DAL3 product (Cys-Ile-Ile-Ile) are homologous to those (CAAX) previously shown to be the primary structural signal for post-translational farnesylation of yeast RAS protein and mating factor. This modification is reported to be responsible for membrane localization of proteins containing it. The upstream region of DAL3 contains six copies of a sequence that is homologous to the positively acting DAL UASNTR reported to be required for transcriptional activation of the DAL5 and DAL7 genes. Missing from the DAL3 upstream region were any sequences related to those shown to be required for a DAL7 response to inducer, the UIS element. This correlates with the previous report that DAL3 expression is independent of the allantoin pathway inducer.

Amidine-Lyases↗

The allantoinase (DAL1) gene of Saccharomyces cerevisiae.

The allantoinase (DAL1) gene from Saccharomyces cerevisiae has been cloned, sequenced, and found to encode a 472 amino acid protein with a Mr of 52,028. DAL1 is expressed in an inducer-independent manner in strain M970 (sigma 1278b genetic background) and modestly responds to mutation of the dal80 locus. Expression was also sensitive to nitrogen catabolite repression (NCR). Correlated with these expression characteristics, the upstream region of DAL1 contained five copies of a sequence that is homologous to the DAL UASNTR element previously shown to be required for transcriptional activation and NCR sensitivity of the DAL5 and DAL7 genes. Missing from the DAL1 5' flanking region were any sequences with significant homology to the DAL7 UIS element required for response to inducer. These observations further support the roles of UASNTR and DAL7 UIS in the regulation of allantoin pathway gene expression.

Amidohydrolases↗