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P Cuevas

Publications and source records attributed to P Cuevas.

At least 19 recordsLinked to original sources
PubMed1992

Single-step purification on DEAE-sephacel of recombinant polypeptides produced in Escherichia coli.

We describe a method for the purification of recombinant proteins based upon the selective interaction of the choline-binding domain of the pneumococcal murein hydrolase and tertiary amines. Proteins of interest, fused to the binding domain by a peptide linker, containing the cleaving sequence recognized by blood coagulation factor Xa, can either be assayed for biological activities in vitro and in vivo or have the binding moiety removed to yield a totally unmodified form, suitable for clinical and functional studies. The method can also be applied to the production of low molecular mass peptides. The principle of the technique is illustrated with acidic fibroblast growth factor and with a neuropeptide-like fragment of ten amino acids contained within its sequence.

3T3 Cells↗
PubMed1992

Fibroblast growth factor injected in cerebral ventricles does not decrease mean arterial blood pressure.

Intracerebroventricular injections of acidic fibroblast growth factor (aFGF) in rats did not elicit any change in the mean arterial blood pressure, and also did not appreciably affect the hypotensive effect of intravenous injections of aFGF. These observations are of general clinical interest, since such effects could constitute important drawbacks for the therapeutic applications of FGF.

Animals↗
PubMed1991

Hypotensive activity of fibroblast growth factor.

Acidic and basic fibroblast growth factors (FGFs) are members of a family of proteins that are broad-spectrum mitogens, have diverse hormone-like activities, and function in tumorigenesis. FGF's ability to raise the concentration of intracellular calcium ion suggests that FGF could induce the synthesis of endothelium-derived relaxing factor (EDRF) and consequently vasodilation. Systemic administration of FGF decreased arterial blood pressure. This effect was mediated by EDRF and by adenosine triphosphate-sensitive potassium ion channels. The hypotensive effect of FGF was segregated from its mitogenic activity by protein engineering. These results extend the range of FGF autocrine activities and potential therapeutic applications, emphasize the role of endothelium as an arterial blood pressure--regulating organ, and provide insight on the structural basis of FGF functions.

Animals↗
PubMed1991

Expression of basic fibroblast growth factor and its receptor in the rat subfornical organ.

Basic fibroblast growth factor (FGF), its mRNA and the mRNA that encodes for its receptor have all been localized in the rat subfornical organ (SFO). Basic FGF is widely distributed throughout the SFO; it is present in neurons, in the vascular basement membrane of lateral blood vessels (but not those within the SFO) and in ependymal cells surrounding the SFO. Results of in situ hybridization show that the expression of basic FGF mRNA is detected throughout the organ. Similarly, the expression of flg, the gene for the putative basic FGF receptor, can also be detected in the SFO. The results all support the possibility that this growth factor may modulate the known physiological functions of the SFO.

Animals↗
PubMed1991

Immunohistochemical localization of basic fibroblast growth factor in ependymal cells of the rat lateral and third ventricles.

The presence of basic fibroblast growth factor (bFGF) in the third and the lateral ventricular ependyma of the rat has been investigated under the light microscope using a polyclonal antibody against bFGF, through the unlabelled peroxidase-antiperoxidase procedure; bFGF-immunoreactivity (bFGF-IR) is observed in the entire ependymal cell layer of ventricles. Also ependymal tanycytes within the inferior portion of the wall and floor of the third ventricle show bFGF-IR. Tanycytic processes are in close contact with hypothalamic capillaries. The present study strongly suggests that brain ependymal bFGF plays unidentified roles unrelated to its angiogenic or mitogenic capacities.

Animals↗
PubMed1991

Vascular response to basic fibroblast growth factor when infused onto the normal adventitia or into the injured media of the rat carotid artery.

Immunohistochemical techniques localize basic fibroblast growth factor (FGF) in endothelial and smooth muscle cells of the common carotid artery. Thus, we studied the effect in rats of basic FGF infused for 14 days onto the adventitia or into the media in vivo. In untreated rats, the adventitial layer is uniform, and few vessels are observed in cross sections (mean +/- SEM is 0.351 +/- 0.16 capillaries/field at a magnification of x 480). Whereas saline infusion increases the mean number of vasa vasorum to 2.73 +/- 0.011 capillaries/field (p less than 0.01), basic FGF (1 ng/microliter/hr) increases the capillary number to 13.4 +/- 0.67 capillaries/field. The effects are local and restricted to the site of delivery; no cell proliferation is observed even 2 mm from the site of infusion. There is also no evidence of the infiltration of macrophages and monocytes. In an effort to determine the effect of basic FGF in the media, a small longitudinal (1-mm) incision was made in the adventitia, and saline or basic FGF (1 ng/microliter/hr) was infused for 14 days into the arterial wall. Under these conditions, basic FGF is a potent inducer of smooth muscle cell proliferation in the vascular wall as well as of new capillaries. In these instances, however, the capillaries formed are thick-walled. The results support the hypothesis that basic FGF may be contributing to the growth and maintenance of the vasa vasorum and of vascular smooth muscle cells.

Animals↗
PubMed1990

Ischemic reperfusion injury in rabbit spinal cord: protective effect of superoxide dismutase on neurological recovery and spinal infarction.

The potential role of superoxide dismutase (SOD), a specific superoxide anion radical scavenger, in treating spinal cord ischemia was investigated in rabbits subjected to aortic occlusion for 20 min. SOD treatment, targeted to the early reperfusion period, reduced both motor dysfunction and incidence of spinal infarcts at 7 days after ischemia. Present results suggest that oxygen-derived free radicals play a role in the pathogenesis of infarcts developing in the spinal cord after ischemia and reperfusion injuries.

Animals↗
PubMed1989

Administration of bovine superoxide dismutase prevents sequelae of spinal cord ischemia in the rabbit.

The contribution of free radical-mediated reperfusion injury to the ischemic damage caused by total arterial occlusion has been investigated in a model of transient spinal cord ischemia in the rabbit. Spinal cord ischemia was produced in 20 anaesthetized rabbits by temporary luminal occlusion (20 min) of the abdominal aorta below the renal arteries. Superoxide dismutase (5 mg/kg) (10 animals) was infused before and during reperfusion below aortic occlusion using an infusion pump that infused the enzyme through the contralateral femoral artery. Control (10 animals) received sterile saline with the same procedure. In this later group, 4 animals developed paraplegia, 4 were paretic and only 2 were normal. However, in the treated group, 6 animals were normal while 3 were paretic and only one appeared paralyzed. We conclude that: a) oxygen free radicals generated during reperfusion are involved in producing the ischemic injury, and b) the ischemic spinal cord injury is prevented by superoxide dismutase.

Animals↗
PubMed1988

Basic fibroblast growth factor (FGF) promotes cartilage repair in vivo.

Although it has been clearly established that basic fibroblast growth factor (FGF) is a potent mitogen for chondrocytes in vitro, there is little evidence that it can stimulate this cell type in vivo. In an effort to address this problem, we examined the effect of an intraarticular administration of basic FGF. Alzet osmotic pumps delivering the mitogen to the site of injury promotes the healing of intra-chondrial lesions by stimulating chondrocyte proliferation and the formation of extracellular matrix. The observation that chronic infusions of basic FGF can elicit a repair response at the site of injury suggests that this growth factor may have therapeutic applications that extend beyond its capacity to induce neovascularization. The results also suggest that one of the ways that the perichondrium mediates cartilage repair may be by the local production of FGF-like mitogens.

Animals↗
PubMed1988

Freeze-fracture cytochemistry of cholesterol content in neuronal plasma membrane following cerebral ischaemia.

Minor changes on the neuronal perikaryon cytoplasm and large alterations in neuronal processes have been demonstrated with transmission electron microscopy after 60 minutes of focal and selective cerebral ischaemia. The distribution of cholesterol in neuronal plasma membrane and perikarya plasmalemma of normal and ischaemic caudate nucleus was investigated with the polyene antibiotic filipin, a morphological probe for membrane cholesterol domains in freeze-fracture replicas. After filipin incubation of prefixed vibratome slices, filipin-cholesterol complexes appeared as 20-30 proturberances and pits on P- and E-faces. Distinct patterns of filipin-cholesterol complexes were found in non-ischaemic and ischaemic neuronal membrane. The filipin-treated specimens showed a 35-40% drop in cholesterol content in the neuronal plasmalemma one hour after cerebral ischaemia.

Animals↗
PubMed1987

An in vivo model for study of the angiogenic effects of basic fibroblast growth factor.

We have investigated the angiogenic effects of basic fibroblast growth factor following its implantation in slow release beads under the kidney capsule. The presence of basic fibroblast growth factor in the subcapsular space induced a marked angiogenic response maximal at 1 microgram dose per kidney. Histological examination at the site of treatment failed to reveal evidence of an inflammatory response, thus supporting the observation that basic fibroblast growth factor alone can stimulate in vivo neovascularization. Beads pretreated with saline or with human growth hormone had no angiogenic effect. Because of the readily accessible location in the retroperitoneal space, the ease of drug delivery, and the marked vascular proliferation seen in response to FGF, our results suggest that the kidney capsule is an excellent model for study of the physiological role played by FGF and related peptides in promoting angiogenesis in vivo.

Animals↗
PubMed1987

Immunohistochemical detection of inhibin in the gonad.

Antiserum to inhibin was produced in rabbits by immunization with a synthetic [Tyr30]alpha-chain(1-30)NH2 fragment of porcine inhibin coupled to bovine serum albumin, and the elicited antiserum was used in conjunction with the avidin-biotin immunoperoxidase procedure to localize inhibin-reactive cells in various rat tissue preparations. In the testes, only the Sertoli cells revealed immunoreactivity with the antiserum. Intense staining was also observed in ovarian follicular granulosa cells but not in the theca layer outside the basement membrane. In addition, the luteal cells in the corpus luteum were also stained by the antiserum. The positive staining in the gonadal tissues could be blocked completely by pre-adsorbing the serum with either the synthetic peptide or native inhibin. Immunostaining was not detected in brain, pituitary, thymus, stomach, pancreas, kidney and adrenal section, thus confirming that inhibin is a polypeptide originating only from specific cells of the gonad.

Animals↗
PubMed1987

Adrenergic innervation of human middle cerebral artery. Ultrastructural observations.

The innervation of the human middle cerebral artery is studied with transmission electron microscopy using the chromaffin reaction technique. Adrenergic nerve fibers and related terminals, with granular electrodense neurotransmitter vesicles, are confined to the tunica adventitia-tunica media transitional zone and the outer layers of the media. These findings may indicate the presence of an adrenergic vasoconstrictor system for circulation control in the human middle cerebral artery.

Cerebral Arteries↗
PubMed1987

Intramembranous cytochemistry: a new morphological technique for studying cholesterol in the astrocyte plasma membrane of ischemic brain cells.

The distribution of cholesterol in the astrocytic plasma membranes of normal and ischemic caudate nucleus in the cat was investigated with the polyene antibiotic, filipin, a morphological probe for membrane cholesterol domains in freeze-fracture tissue replicas. After filipin incubation of prefixed vibratome slices, filipin-cholesterol complexes appeared as 20 to 30 nm protuberances and pits on P- and E-faces of the replicas. Distinct patterns of filipin-cholesterol complexes were found on nonischemic and ischemic astrocyte membranes. The filipin-treated specimens showed an 80% decrease in astroglial plasmalemma of cholesterol 1 hour after cerebral ischemia.

Animals↗
PubMed1986

Lack of effect of compound 48/80 on human umbilical vein endothelium.

Human umbilical veins have been treated with compound 48/80, a histamine liberator factor, in order to obtain information about the histamine content in the Weibel-Palade bodies. Ultrastructural observations show a lack of effect in the morphology of this organelle. Our findings suggest that the Weibel-Palade bodies are not histamine storage organelles in human umbilical vein.

Endothelium↗