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Biomedical subjects

J L Ward

Publications and source records attributed to J L Ward.

At least 19 recordsLinked to original sources
PubMed2001

Characterization of nucleoside transport systems in cultured rat epididymal epithelium.

The nucleoside transport systems in cultured epididymal epithelium were characterized and found to be similar between the proximal (caput and corpus) and distal (cauda) regions of the epididymis. Functional studies revealed that 70% of the total nucleoside uptake was Na(+) dependent, while 30% was Na(+) independent. The Na(+)-independent nucleoside transport was mediated by both the equilibrative nitrobenzylthioinosine (NBMPR)-sensitive system (40%) and the NBMPR-insensitive system (60%), which was supported by a biphasic dose response to NBMPR inhibition. The Na(+)-dependent [(3)H]uridine uptake was selectively inhibited 80% by purine nucleosides, indicating that the purine nucleoside-selective N1 system is predominant. Since Na(+)-dependent [(3)H]guanosine uptake was inhibited by thymidine by 20% and Na(+)-dependent [(3)H]thymidine uptake was broadly inhibited by purine and pyrimidine nucleosides, this suggested the presence of the broadly selective N3 system accounting for 20% of Na(+)-dependent nucleoside uptake. Results of RT-PCR confirmed the presence of mRNA for equilibrative nucleoside transporter (ENT) 1, ENT2, and concentrative nucleoside transporter (CNT) 2 and the absence of CNT1. It is suggested that the nucleoside transporters in epididymis may be important for sperm maturation by regulating the extracellular concentration of adenosine in epididymal plasma.

Animals↗
PubMed2000

Kinetic and pharmacological properties of cloned human equilibrative nucleoside transporters, ENT1 and ENT2, stably expressed in nucleoside transporter-deficient PK15 cells. Ent2 exhibits a low affinity for guanosine and cytidine but a high affinity for inosine.

We stably transfected the cloned human equilibrative nucleoside transporters 1 and 2 (hENT1 and hENT2) into nucleoside transporter-deficient PK15NTD cells. Although hENT1 and hENT2 are predicted to be 50-kDa proteins, hENT1 runs as 40 kDa and hENT2 migrates as 50 and 47 kDa on SDS-polyacrylamide gel electrophoresis. Peptide N-glycosidase F and endoglycosidase H deglycosylate hENT1 to 37 kDa and hENT2 to 45 kDa. With hENT1 being more sensitive, there is a 7000-fold and 71-fold difference in sensitivity to nitrobenzylthioinosine (NBMPR) (IC(50), 0.4 +/- 0.1 nM versus 2.8 +/- 0.3 microM) and dipyridamole (IC(50), 5.0 +/- 0.9 nM versus 356 +/- 13 nM), respectively. [(3)H]NBMPR binds to ENT1 cells with a high affinity K(d) of 0.377 +/- 0.098 nM, and each ENT1 cell has 34,000 transporters with a turnover number of 46 molecules/s for uridine. Although both transporters are broadly selective, hENT2 is a generally low affinity nucleoside transporter with 2.6-, 2.8-, 7. 7-, and 19.3-fold lower affinity than hENT1 for thymidine, adenosine, cytidine, and guanosine, respectively. In contrast, the affinity of hENT2 for inosine is 4-fold higher than hENT1. The nucleobase hypoxanthine inhibits [(3)H]uridine uptake by hENT2 but has minimal effect on hENT1. Taken together, these results suggest that hENT2 might be important in transporting adenosine and its metabolites (inosine and hypoxanthine) in tissues such as skeletal muscle where ENT2 is predominantly expressed.

Biological Transport↗
PubMed1999

Nucleoside transport in human colonic epithelial cell lines: evidence for two Na+-independent transport systems in T84 and Caco-2 cells.

RT-PCR of RNA isolated from monolayers of the human colonic epithelial cell lines T84 and Caco-2 demonstrated the presence of mRNA for the two cloned Na+-independent equilibrative nucleoside transporters, ENT1 and ENT2, but not for the cloned Na+-dependent concentrative nucleoside transporters, CNT1 and CNT2. Uptake of [3H]uridine by cell monolayers in balanced Na+-containing and Na+-free media confirmed the presence of only Na+-independent nucleoside transport mechanisms. This uptake was decreased by 70-75% in the presence of 1 microM nitrobenzylthioinosine, a concentration that completely inhibits ENT1, and was completely blocked by the addition of 10 microM dipyridamole, a concentration that inhibits both ENT1 and ENT2. These findings indicate the presence in T84 and Caco-2 cells of two functional Na+-independent equilibrative nucleoside transporters, ENT1 and ENT2.

Caco-2 Cells↗
PubMed1997

Synthesis of cytokinin glucuronides for the selection of transgenic plant cells.

Glucuronide derivatives of cytokinins have been synthesized for use as agents for the selection of plant cells transformed with a beta-glucuronidase (GUS) gene. In this selection system, the GUS gene functions as both a selectable, as well as a screenable gene. GUS liberates active cytokinin from inactive cytokinin glucuronides which then stimulates growth and regeneration of the transformed cells. The frequently used cytokinin N6-benzyladenine was conjugated to glucuronic acid at N-3 or at N-9 but only the former was a substrate for GUS. The glucuronide of isopentenyladenine was also made, by coupling at the N-3. This compound was readily hydrolysed by GUS.

Cell Division↗
PubMed1995

Absorption pharmacokinetics of atenolol in patients with the Marfan syndrome.

The negative inotropic and chronotropic effects of beta-blocker therapy have been reported to reduce morbidity and mortality in patients with Marfan syndrome; however, little is known about the pharmacokinetics of atenolol after oral administration of multiple doses to patients with the Marfan syndrome. We studied the pharmacokinetics of atenolol in 13 such patients aged 18.7 +/- 2.9 years who were receiving 1.78 +/- 0.58 mg/kg/day (70.1 +/- 20.3 mg/m2/day) of atenolol for 6 weeks or longer. Mean +/- SD percentage change in baseline heart rate after the administration of atenolol was -18.03 +/- 16.59% and mean +/- SD percentage change in exercise heart rate after atenolol was -33.22 +/- 14.75% (P < .01). Six to 8 atenolol serum concentrations were collected in each patient during a 12-hour dosing interval and were determined by high-performance liquid chromatography with ultraviolet detection. Serum atenolol concentrations at 0 (123 +/- 70 micrograms/L) and 12 (116 +/- 66 micrograms/L) hours were within 20% of each other and were thus assumed to be at steady-state. A one-compartment, steady-state pharmacokinetic model with first-order absorption and elimination was fitted to the concentration-time data for each patient using nonlinear regression. Maximal concentration was 343 +/- 120 micrograms/L, and the mean half-life was 4.72 hours.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1994

Partial typhlectomy and ileocolostomy for treatment of nonreducible cecocolic intussusception in a horse.

Cecocolic intussusception was detected in a 2-year-old male Standardbred horse with a 3-day history of signs of intermittent colic. The entire cecum, which was located within the lumen of the right ventral colon, was edematous and necrotic, and could not be manually reduced. A colotomy was made, and partial typhlectomy was performed. An ileocolostomy also was performed. To prevent eversion of the cecal base, the site of invagination into the colon was oversewn. Several postoperative complications were treated, including peritonitis, thrombophlebitis, anemia, and hypoproteinemia, and 1 year after surgery, the horse was healthy and in training.

Animals↗
PubMed1994

Evaluation of abomasal outflow diversion as an experimental model of hypochloremic, hypokalemic metabolic alkalosis in lactating cows.

Four adult, lactating dairy cows were subjected to diversion (loss) of gastric contents through a T-shaped cannula placed in the cranial part of the duodenum just distal to the pylorus. Diversion was continued for 10 to 12 hours, at which point the cows were very weak and depressed. The volume of effluent during this period ranged from 37.3 to 46.8 L, with the largest volume being produced during the first four hours. All cows became dehydrated, with mean packed cell volume and total plasma protein concentration increasing 30% and 19.6%, respectively, but with only a slight increase in plasma creatinine concentration. Plasma Cl- concentrations decreased from a mean of 97.3 mEq/L at the beginning of diversion to a mean of 87.2 mEq/L at eight hours. This was followed by a plateau or slight increase in concentrations over the final hours of diversion. Plasma K+ concentration followed a similar pattern, decreasing from a mean of 3.9 mEq/L to a mean of 2.94 mEq/L at six hours, followed by increasing values until termination of diversion. No changes in plasma Na+ concentration were noted, except for a mild decrease in one cow. Plasma calcium concentrations decreased significantly, reaching 6.6 +/- 0.6 mEq/L at the end of diversion. Venous pH, plasma HCO3- concentration, and plasma base excess concentration increased during the first four to eight hours of diversion, followed by a gradual decline. Although a mild hypochloremic metabolic alkalosis resulted from diversion of abomasal outflow in all cows, substantiated by a mild increase in plasma strong ion difference, the changes observed were not as great as expected.(ABSTRACT TRUNCATED AT 250 WORDS)

Abomasum↗
PubMed1993

Repair of sacral fracture in two dairy cattle.

Sacral fractures in 2 dairy cattle were repaired surgically for cosmetic reasons. A heifer had a Salter-Harris type-I fracture of the fifth sacral vertebra, which was repaired with a 4.5-mm narrow dynamic compression plate. A cow had complete fracture of the fourth sacral body, which was repaired with 2 extra-large plastic spinous process plates. Both fixations were successful in restoring the dorsal contour of the sacrococcygeal region.

Animals↗
PubMed1993

Comparison of 0.9, 3.6, and 7.2% NaCl for correction of experimentally induced hypochloremic, hypokalemic metabolic alkalosis in sheep.

Nine adult female sheep were each surgically fitted with an Ivan and Johnston reentrant cannula in the cranial part of the duodenum just distal to the pylorus. By diversion (loss) of abomasal outflow, this model has been shown to consistently induce hypochloremic, hypokalemic metabolic alkalosis, accompanied by hyponatremia and dehydration. Each sheep was subjected to 3 treatment trials, each preceded by a 24-hour prediversion period, and a diversion period during which a syndrome of hypochloremia (68 +/- 2 mEq/L), hypokalemia, hyponatremia, and metabolic alkalosis was induced. Development of this syndrome was attributable to losses of large amounts of acid and electrolytes in the abomasal effluent. Mean total electrolyte contents of the effluent were: Cl-, 650 +/- 27 mEq; Na+, 388 +/- 23 mEq; and K+, 123 +/- 12 mEq, with total volume loss ranging from 3.6 to 10.0 L of gastric contents and pH ranging from 3 to 5. Decreases in plasma electrolyte concentrations also can be attributed to decreased intake, because anorexia developed shortly after the onset of diversion. Electrolyte losses in urine during diversion were minimal for Cl- (mean +/- SEM, 12.0 +/- 5.1 mEq), but were greater for Na+ (124.2 +/- 14.5 mEq) and K+ (185.1 +/- 31.2 mEq). Treatments consisted of 0.9% NaCl (300 mosm/L), 3.6% NaCl (1,200 mosm/L), and 7.2% NaCl (2,400 mosm/L) administered over a 2-hour period, with the administered volume determined by the estimated total extracellular fluid Cl- deficit. Significant difference was not found among treatments, with all solutions resulting in return of clinicopathologic and physical variables to prediversion values within 12 hours of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Abomasum↗
PubMed1992

Chronic frontal sinusitis in dairy cattle: 12 cases (1978-1989).

Chronic frontal sinusitis in 12 dairy cattle most often was associated with a history of dehorning, in which the sinus was entered (67%), or with respiratory tract disease (25%). The most common organisms isolated were Actinomyces pyogenes and Pasteurella multocida. Signs of infection did not develop for months in some cattle and were often intermittent. The most common clinical signs included anorexia, lethargy, fever, frontal bone distortion, exophthalmos, abnormal posture, nasal discharge, and neurologic abnormalities. Treatment consisted of trephination at 2 sites, drainage and lavage of the sinus cavity, and administration of antibiotics and analgesics. Eight cattle responded well to treatment and were discharged, but 4 others had signs of CNS involvement and died or were euthanatized. Trephination of the frontal sinus cavity at carefully chosen sites and antibiotic treatment are indicated when sinusitis is suspected. Drainage of the sinus cavity is imperative to avoid extension of the infection into the CNS.

Animals↗
PubMed1992

Medullary thyroid carcinoma: Australian experience with genetic testing.

Linkage analysis has been performed in four pedigrees with multiple endocrine neoplasia type 2A (MEN 2A) or familial medullary thyroid carcinoma (MTC) using pericentromeric chromosome 10 probes. Important information regarding carrier status has been provided in 10 individuals, many of whom would not have been identified by pentagastrin stimulation testing. We have also used pulsed field gel electrophoresis (PFGE) to link the probes H4.IRBP and pMCK2 to a 150 kb fragment. Using PFGE, no evidence was found in DNA from lymphocytes of a major DNA rearrangement in two individuals affected with MEN 2A and an individual with MEN 2B compared with normals. Metastatic MTC from one patient has been used to generate a cDNA library which will be used to screen for candidate MEN 2A and MEN 2B gene(s).

Carcinoma↗
PubMed1988

Alteration of morphogenesis by the v-myc oncogene in transplants of mammary gland.

To see if individual oncogenes can alter three-dimensional growth in vivo, we have inserted the v-myc oncogene into transplants of mouse mammary gland. Primary cultures of mammary epithelial cells were infected with helper-free retrovirus to insert v-myc oncogenes, and transplanted into cleared mammary fat pads (mammary glands from which the natural epithelium had been removed) of host mice. Uninfected transplants, and transplants infected with retrovirus constructs that carry no oncogene, grew to form an epithelial 'tree' resembling normal mammary gland, as expected. Transplants infected with retroviruses carrying v-myc oncogenes grew to form a characteristic, abnormal (hyperplastic) pattern in which the ducts were more densely packed than normal. Integration of the retrovirus in the transplants was demonstrated. The effect of the oncogene was local, not systemic, as some transplants showed adjacent areas of normal and hyperplastic growth. Thus v-myc can alter morphogenesis without growth becoming disorganized.

Animals↗