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Biomedical subjects

D Burks

Publications and source records attributed to D Burks.

7 recordsLinked to original sources

Recombinant mouse ZP3 inhibits sperm binding and induces the acrosome reaction.

Mammalian fertilization involves interactions of sperm surface receptors with ligands of the zona pellucida, an extracellular matrix surrounding the ovulated egg. In mouse, the zona is composed of three glycoproteins. One of them, ZP3, participates in primary sperm binding and in the subsequent triggering of the sperm's acrosome reaction. Considerable evidence suggests that carbohydrate determinants of ZP3 are responsible for binding to sperm and may be important for acrosomal exocytosis. A full-length cDNA encoding mouse ZP3 was assembled and cloned into expression vectors that contained either a cytomegalovirus (CMV) or a vaccinia (P11) promoter. Mouse L-929 cells were stably transformed with the pZP3-CMV constructs, and green monkey CV-1 cells were infected with a recombinant vaccinia virus containing ZP3. rZP3 was affinity purified from culture media and detected on Western blots as a single 60- to 70-kDa band, which differed in molecular weight from native ZP3 (mean, 83 kDa). Nevertheless, rZP3 is biologically active. rZP3 decreases sperm-zona binding with a potency equivalent to that of native zona pellucida and, like native ZP3, rZP3 triggers acrosomal exocytosis in capacitated mouse sperm. Thus, rZP3 isolated from both rodent and primate cells appears to contain those carbohydrate and protein structures necessary for ZP3's dual role in fertilization.

Animals

Evidence that cyclic adenosine 3',5'-monophosphate-dependent protein kinase activation causes pig ovarian granulosa cell differentiation, including increases in two type II subclasses of this kinase.

Agents that elevated intracellular cyclic adenosine 3',5'-monophosphate (cAMP) caused a 3- to 10-fold increase in the luteinizing hormone (LH) receptor level and in progesterone biosynthesis in primary cultures of pig ovarian granulosa cells. Associated with these effects was a 2- to 4-fold increase in the total activity of the catalytic subunit of cAMP-dependent protein kinase in the tissue. From quantitation by [3H]cAMP binding and changes in the specific labeling with the photoaffinity analog [32P]-8-azido-cAMP, these agents were found to cause a concomitant 5- to 15-fold increase in two isoforms of the type II R-subunit (Mr = 54,000 and 56,000) of the protein kinase. Since the two intrasubunit cAMP binding sites of the protein kinase have been found to be positively cooperative, the addition of a combination of an analog selective for site 1 and an analog selective for site 2 causes synergistic increases in protein kinase activation in vitro and synergistic increases in intact cell responses if mediated by the cAMP-dependent protein kinase. In the present study, the addition of such a combination of site 1- and site 2-selective analogs to granulosa cells caused a synergistic increase in LH receptor induction and progesterone production. For both responses, synergism did not occur when two analogs selective for the same site were combined. The results indicated that these responses are mediated by either of the two major isozyme types of cAMP-dependent protein kinase.

Animals

Urethral cyst in a man.

We report a case of a bulbous urethral cyst in a man. This rare lesion, probably arising from Cowper's gland, was treated successfully by endoscopic excision of the cyst wall.

Adult

Co-localization of enkephalin and cholecystokinin in discrete areas of rat brain.

A double-label immunofluorescence technique was used to demonstrate that immunoreactivities for the functionally antagonistic neuropeptides enkephalin and cholecystokinin octapeptide (CCK) are co-localized within individual neurons and processes in discrete areas of rat midbrain and forebrain. Coexistence was most prominent within varicose pericellular axons extending from the periaqueductal gray matter to a field overlying the medial lemniscus, axons and terminal-like puncta in the central medial, paracentral, interanterodorsal and ventral anterior thalamic nuclei, and perikarya and proximal axonal fragments in layers II and III of neo- and allocortex, and in the anterior olfactory nucleus. The former two systems of axons lie in areas of spinothalamic tract termination. These data suggest that some of the antagonism of opioid analgesia by CCK occurs at the synaptic level in nociceptive areas of brain-stem and thalamus where CCK and enkephalin are co-localized and presumably co-released.

Animals

Vicarious excretion of contrast medium in patients without azotemia.

Although excretion of urographic iodinated contrast agents via the biliary and gastrointestinal tract is not uncommon in patients with renal insufficiency, such vicarious excretion is unusual in the presence of normal renal function. The observation of such vicarious excretion in 2 patients with acute unilateral ureteral obstruction and no azotemia is reported in conjunction with review of the appropriate literature and suggestion of possible etiologies.

Adult

Effects of coronary flow reduction on capillary-myocardial exchange in dogs.

The effects of coronary flow reduction on tracer capillary permeability surface area (PS) and distribution volumes were studied in open-chest dog preparations. A mixture of 51Cr-labeled red blood cells, 125I-labeled albumin, [14C]sucrose, and tritiated water (3H2O) was introduced into a shunt connecting the carotid and left anterior descending coronary arteries. Sampling from the coronary sinus produced a multiple indicator curve from which [14C]sucrose PS and 3H2O volumes were computed. Curves were observed in control situations and after the cannula was partially clamped. In six dogs, cardiac lymph was collected and analyzed for total protein. Paired comparison of control and flow-restricted indicator curves showed that flow reduction decreased the absolute values of PS and tracer volumes. The ratio of sucrose volume to weight of perfused tissue increased with flow reduction. The ratio of sucrose PS to weight of perfused tissue increased with moderate flow reduction and decreased with severe flow reduction. The results suggest that flow reduction has two effects which competively affect exchange: 1) flow restriction reduces surface area by capillary derecruitment, and 2) the remaining functional capillaries appear to undergo an increase in permeability to small molecules.

Animals