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Biomedical subjects

M Campos

Publications and source records attributed to M Campos.

At least 19 recordsLinked to original sources

Photorefractive keratectomy for severe postkeratoplasty astigmatism.

We performed cylindric corneal ablations with the excimer laser on 12 patients to correct severe, disabling astigmatism after keratoplasty. In some patients, an additional ablation was performed to correct myopia. Patients were followed up for an average of eight months (range, six to 14 months). Uncorrected visual acuity improved in nine patients, and nine of the 12 patients had a decrease in refractive cylinder at last follow-up. The mean preoperative refractive cylinder was 7.0 +/- 3.6 diopters, which decreased to a mean of 3.1 +/- 2.6 diopters at one month (P = .0003) and 4.3 +/- 2.9 diopters at last follow-up (P = .03). Keratometric astigmatism decreased from 7.5 +/- 3.9 diopters preoperatively to 5.2 +/- 3.9 diopters at the last follow-up (P = .001). Mean spherical equivalent was reduced from -7.4 +/- 4.2 diopters preoperatively to -3.3 +/- 4.4 diopters postoperatively (P = .003). Postoperative corneal haze, when present, did not reduce visual acuity. Excimer laser superficial keratectomy thus appears to be safe when used for postkeratoplasty ametropia, although substantial regression may limit its effectiveness in some patients.

Adult

Role of the endothelial system in Bay K 8644 enantiomer and nifedipine vasomodulator action in rat aorta.

The potential importance of the endothelial system in regulating the effects of (-)-Bay K 8644 (0.1 microM), (+)-Bay K 8644 (0.1 microM) and nifedipine (10 nM) on resting tension, on contractile responses to noradrenaline (NA) and Ca2+ (in a Ca(2+)-free high-K+ solution), and on basal, NA-induced and K(+)-induced 45Ca2+ uptake, was investigated in rat aorta rings. Mechanical removal of endothelium considerably potentiated the contractile response induced by NA in standard medium and by Ca2+ in Ca(2+)-free high-K+ (15 mM) medium, but did not modify the response induced by Ca2+ in Ca(2+)-free high-K+ (55 mM) medium or by NA in Ca(2+)-free medium. Furthermore, the basal 45Ca2+ uptake and that induced by NA (10 microM) or KCl (15 and 55 mM) were similar in endothelium-rubbed and intact rings. (-)-Bay K 8644 (0.1 microM) shifted the NA and Ca2+ concentration-response curves to the left with potentiation of the maximal contraction. However, (+)-Bay K 8644 (0.1 microM) and nifedipine (10 nM) caused a shift to the right, with depression of the maximal contraction. The NA concentration-response curves, and those of Ca2+ in Ca(2+)-free high-K+ (55 mM) medium, were affected by the drugs to similar extents, and were not modified by the presence or absence of endothelial cells. The drugs tested did not affect resting tension. Basal 45Ca2+ uptake was not modified by either nifedipine or the Bay K 8644 enantiomers.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy

Corneal sensitivity after photorefractive keratectomy.

Corneal anesthesia or hypesthesia can complicate refractive surgical procedures such as epikeratophakia and radial keratotomy. An esthesiometer was used to measure the corneal sensitivity in unoperated-on corneas and fellow corneas after excimer laser photorefractive keratectomy. Decrease in corneal sensitivity was noted within six postoperative weeks, with mean sensitivity being 75.2% +/- 13.3% of normal. Within the first three postoperative months, the patients operated on for correction of compound astigmatism recovered 95.7% +/- 5.3% of the corneal sensitivity, whereas the patients operated on for correction of severe myopia recovered 86.2% +/- 11.2% (P = .07). None of the patients had delayed epithelial healing or recurrent corneal erosions during the time of decreased corneal sensitivity. In otherwise normal myopic eyes, photorefractive keratectomy measurably reduced corneal sensitivity for several postoperative weeks.

Adult

Effects of magnesium and ATP on pre-steady-state phosphorylation kinetics of the Na+,K(+)-ATPase.

The aim of the present work was to elucidate the role played by ATP and Mg2+ ions in the early steps of the Na+,K(+)-ATPase cycle. The approach was to follow pre-steady-state phosphorylation kinetics in Na(+)-containing K(+)-free solutions under variable ATP and MgCl2 concentrations. The experiments were performed with a rapid mixing apparatus at 20 +/- 2 degrees C. The concentrations of free and complexes species of Mg2+ and ATP were calculated on the basis of a dissociation constant of 0.091 +/- 0.004 mM, estimated with Arsenazo III under identical conditions. A simplified scheme were ATP binds to the ENa enzyme, which is phosphorylated to MgEPNa and consequently dephosphorylated returning to the ENa form, was used. In the absence of ADP and phosphate four rate constants are relevant: k1 and k-1, the on and off rate constants for ATP binding; k2, the transphosphorylation rate constant and k3, the constant that governs the dephosphorylation rate. The values obtained were: k1 = 0.025 +/- 0.003 microM-1 ms-1 for both free ATP and ATPMg; k-1 = 0.038 +/- 0.004 ms-1 for free ATP and 0.009 +/- 0.002 ms-1 for ATPMg; k2 = 0.199 +/- 0.005 ms-1; k3 = 0.0019 +/- 0.0002 ms-1. The model that seems best to explain the data is one where (i) the role of true substrate can be played equally well by free ATP or ATPMg, and (ii) free Mg2+, an essential activator, acts by binding to a specific Mg2+ site on the enzyme molecule.

Adenosine Triphosphate

Structural, functional, and immunological characterization of bovine herpesvirus-1 glycoprotein gl expressed by recombinant baculovirus.

The major glycoprotein complex gl of bovine herpesvirus-1 was expressed at high levels (36 micrograms per 1 x 10(6) cells) in insect cells using a recombinant baculovirus. The recombinant gl had an apparent molecular weight of 116 kDa and was partially cleaved to yield 63-kDa (glb) and 52-kDa (glc) subunits. This processing step was significantly less efficient in insect cells than the analogous step in mammalian cells, even though the cleavage sites of authentic and recombinant gl were shown to be identical. The oligosaccharide linkages were mostly endoglycosidase-H-sensitive, in contrast to those of authentic gl, which has mostly endoglycosidase-H-resistant linkages and an apparent molecular weight of 130/74/55 kDa. Despite the reduced cleavage and altered glycosylation, the recombinant glycoprotein was transported and expressed on the surface of infected insect cells. These surface molecules were biologically active as demonstrated by their ability to induce cell-cell fusion. Fusion was inhibited by three monoclonal antibodies specific for antigenic domains I and IV on gl. Domain I maps to the extracellular region of the carboxy terminal fragment glc and domain IV to the very amino terminus of the glb fragment, indicating that domains mapping in two distinct regions of gl function in cell fusion. Monoclonal antibodies specific for eight different epitopes recognized recombinant gl, indicating that the antigenic characteristics of the recombinant and authentic glycoproteins are similar. In addition, the recombinant gl was as immunogenic as the authentic gl, resulting in the induction of gl-specific antibodies in cattle.

Amino Acid Sequence

Characterization and activation requirements of bovine lymphocytes acquiring cytotoxic activity after interleukin-2 treatment.

Interleukin-2 (IL-2) treatment of cells and generation of non-major histocompatibility complex (MHC)-restricted cytotoxic cells from peripheral blood mononuclear leukocytes (PBML) was studied. Effector-target conjugate assays demonstrated that bovine PBML bound but did not lyse K562, HL60S and HL60R cells unless activated with IL-2. The magnitude of IL-2-activated killing of tumor cells as well as the magnitude of antibody-dependent cellular cytotoxicity depended on the IL-2 concentration. A short treatment (12-18 h) of effector cells with IL-2 was sufficient for development of cytotoxic activity. Withdrawal of IL-2 from the culture resulted in a reduction of cytotoxic activity that could be restored by further addition of IL-2. Cytotoxic activity of IL-2-activated populations obtained after nylon wool or Sephadex G-10 passage, and Percoll gradient centrifugation of PBML suggests that lymphokine-activated killer (LAK) cell activity in PBML is mainly mediated by a non-adherent lymphocyte lacking markers for B-cells. Positive and negative selection experiments using cell sorting confirmed these findings and demonstrated that the cell responsible for LAK cell activity in cattle belongs to a non-monocyte, non-B, CD2+ lymphocyte population. Furthermore, cytotoxic activity could not be generated in CD2+ populations enriched for cells expressing molecules equivalent to human and murine CD4 and CD8. These findings suggest that effector cells mediating non MHC-restricted cytotoxicity in cattle prevail in a population bearing a CD2+, CD4-, CD8- phenotype and that this population depends on the continuous presence of IL-2 for optimal cytotoxic function.

Animals

Multiple administration with interleukin-2 potentiates antigen-specific responses to subunit vaccination with bovine herpesvirus-1 glycoprotein IV.

Interleukin-2 has been described as an effective adjuvant for a number of antigens in different host species. Previously, we demonstrated the adjuvant activity of recombinant bovine IL-2 with a glycoprotein IV (gIV) subunit vaccine from bovine herpesvirus type-1 (BHV-1). In the present study, primary antibody responses were assessed in cattle immunized with either 2 or 50 micrograms of gIV, and treated with multiple doses of IL-2 or combinations of IL-2 and IFN-alpha or IL-2 and IFN-gamma. IL-2 was able to augment significantly antibody responses detected by either ELISA or virus neutralization. More significantly, IL-2 was able to enhance antibody titres in animals immunized with only 2 micrograms gIV to levels similar to those immunized with 50 micrograms gIV in the absence of IL-2. For optimal stimulation, multiple injections of IL-2 and Avridine had to be used in the formulation; other oil adjuvants or IL-2 alone could not induce a primary serum antibody response. Addition of IFN-alpha or IFN-gamma to the IL-2/gIV/Avridine formulation did not affect any of the immune parameters tested. As IFN-alpha is an effective immunoprophylactic agent for infectious bovine rhinotracheitis (IBR), combination vaccine-immunoprophylaxis may become feasible using IL-2 as a co-adjuvant. Thus, extremely low doses of antigen and only one immunization may be an effective vaccine given in combination with interferon prophylactic treatment.

Animals

Effect of urethane on hydralazine-induced tachycardia in rats.

1. In conscious normotensive rats, hydralazine (5-10 mg kg-1 p.o.) produced a dose-related fall in systolic blood pressure, accompanied by a pronounced increase in heart rate. 2. The tachycardia induced by hydralazine (10 mg kg-1 p.o.) in conscious normotensive rats was strongly inhibited after anaesthesia with urethane (1.26 g kg-1 i.p.). 3. In anaesthetized normotensive rats, hydralazine (1 mg kg-1 i.v.) caused a fall in mean blood pressure, accompanied by irregular effects on the heart rate that consisted in a combination of initial tachycardia followed by bradycardia. 4. In pithed rats, hydralazine (1 mg kg-1 i.v.) did not affect mean arterial blood pressure but produced a significant decrease in heart rate. 5. In rat isolated atria, hydralazine (2 mM) produced a positive inotropic/negative chronotropic effect. 6. These results suggest that urethane inhibits the cardiovascular reflex that causes the tachycardia induced by hydralazine in conscious normotensive rats. For this reason, in anaesthetized normotensive rats appear the direct effect of the drug on the heart.

Anesthesia

Effect of Haemophilus somnus on phagocytosis and hydrogen peroxide production by bovine polymorphonuclear leukocytes.

The interactions between bovine polymorphonuclear leukocytes (PMNs) and the bacterium Haemophilus somnus are known to be complex. In this paper, we evaluated the effect of H. somnus on PMN function using a flow cytometric (FC) technique that simultaneously determined the extent of phagocytosis and hydrogen peroxide production by PMNs, as well as using conventional techniques, such as the nitroblue tetrazolium (NBT) and chemiluminescence assays, to analyse the PMN respiratory burst. Results from the FC and chemiluminescence assays demonstrated that in vitro exposure of PMNs to logarithmically growing H. somnus reduced the respiratory burst of PMNs obtained from healthy calves. However, this reduction was not detected by the NBT assay. A decrease in phagocytosis by PMNs could also be shown using the FC assay. In addition, PMNs from calves with acute Hemophilosis (i.e. exposed to H. somnus in vivo) showed reduced activity when compared to PMNs from healthy calves. These in vitro and in vivo observations indicate that the modulation of bovine PMN function by H. somnus may contribute significantly towards the pathogenesis of the disease.

Animals

Corneal wound healing after excimer laser ablation. Effects of nitrogen gas blower.

PURPOSE: To examine the effects of blowing nitrogen gas over the cornea during photorefractive keratectomy. METHODS: Excimer laser ablations for myopia were performed on rabbit corneas with or without the blowing of nitrogen across the surface of the cornea. All eyes underwent a 5-diopter myopic ablation; in 8 eyes, a ring was used to blow nitrogen gas across the cornea, and, in 8 eyes, the same ring was used, but no nitrogen gas was blown. RESULTS: Epithelial healing occurred more rapidly in the eyes that were not treated with the gas (3.8 +/- 1.3 days) than in the gas-treated group (6.1 +/- 0.8 days; P = 0.0025). Corneal haze was greater in the group treated with gas. Results of histologic examination showed the ablated area to have a smoother surface when nitrogen was not blown across the cornea surface. CONCLUSION: Superficial corneal deturgescence produced by the nitrogen gas appears to result in a rougher surface immediately postoperatively with undesirable effects on surface healing, but further studies will be necessary to determine the applicability of these results to humans.

Animals

The role of bovine intraepithelial leukocyte-mediated cytotoxicity in enteric antiviral defense.

The defense of a mucosal surface against viral infection is dependent in part on the leukocyte population resident at that site. In this study, leukocytes from the bovine intestinal epithelium were isolated and assessed for their ability to inhibit in vitro replication of an enteric pathogen, bovine coronavirus (BCV). As well, the intraepithelial leukocytes (IEL) were tested for their ability to mediate different types of cytotoxicity. The IEL were able to inhibit virus replication, and this activity was markedly enhanced by interleukin-2 and tumor necrosis factor. This combination of cytokines has similar effects on IEL-mediated cytotoxicity, which implicated cytotoxicity as a mechanism by which viral replication was limited. The IEL demonstrated enhanced cytotoxic function when compared with lymphocytes isolated from other sites in the gut-associated or systemic immune system. The IEL mediated higher levels of IL-2-activated, antibody-dependent, and lectin-dependent cytotoxicity than did lymphocytes from mesenteric lymph nodes, Peyer's patches, or the spleen. This function may be a reflection of the type of cell recruited to the epithelium, as indicated by the increased prevalence of T cells, and particularly CD8+ cells in the IEL population. Cytokine activation and the presence of a recognition signal, such as antibody, resulted in a synergistic increase in the level of IEL-mediated cytotoxicity. This type of interaction could serve to enhance the efficiency of IEL cytotoxic cells in vivo. Thus IEL-mediated cytotoxicity has the potential to serve as a mechanism of defense to enteric viral infection.

Animals

Comparison of high titer Edmonston-Zagreb, Biken-CAM and Schwarz measles vaccines in Peruvian infants.

In an effort to identify the optimal dose and strain of measles vaccination for early immunization, Peruvian infants were randomly assigned to receive one of three measles vaccines in varying doses at 5 to 6 or 8 to 9 months of age. Edmonston-Zagreb vaccines were significantly (P < 0.001) more immunogenic than equivalent or higher titers of Schwarz or Biken-CAM vaccines as determined by neutralization antibody response 3 months after vaccination. Eighty-two percent of infants who received high titer Edmonston-Zagreb vaccine at 5 to 6 months of age developed protective concentrations of measles antibody, a response rate similar to that observed after standard titer Schwarz (81%) or high titer Biken-CAM vaccine (81%) at 8 to 9 months of age. No significant differences in the rates of fever, rash or other adverse events were noted by vaccine group 10 to 14 days after vaccination. Although the high titer vaccines are more immunogenic in young infants than standard vaccines, long term safety must be assured before these vaccines can be put into widespread use.

Antibodies, Viral

Effects of platelet activating factor on contractions and 45Ca influx induced by noradrenaline and potassium in rat rubbed and intact aorta. Comparison with its hypotensive effect in anaesthetized normotensive rats.

In order to clarify the mechanism of hypotensive activity of platelet activating factor (PAF), the effects of this drug on blood pressure in anaesthetized normotensive rats, on KCl- and noradrenaline-induced 45Ca uptake and contractile responses in rat aorta rings with and without endothelium were studied. PAF (3 micrograms kg-1, i.v.) showed long-lasting hypotensive effects in anaesthetized normotensive rats accompanied by a significant increase in heart rate. PAF (0.1-10 microM) did not relax the contractions induced by noradrenaline (10 microM) or K+ (60 mM) in rubbed or intact rat aorta. PAF did not affect the basal uptake of 45Ca2+ nor that induced by the two vasoconstrictor agents. In experiments in a calcium free medium, PAF (10 microM) had no effect on the noradrenaline- (10 microM) induced contractions. These results suggest that the hypotensive activity of PAF in normotensive anaesthetized rats is not due to a direct effect on rubbed and intact rat aorta rings (acting within the cell or blocking Ca2+ influx through L-type transmembrane calcium channels).

Anesthesia

Molecular chimerization of Pasteurella haemolytica leukotoxin to interleukin-2: effects on cytokine and antigen function.

A chimeric recombinant protein composed of the lktA gene product from Pasteurella haemolytica fused to bovine interleukin-2 (IL-2) was made. The LKT-IL-2 chimera was compared with recombinant bovine IL-2 with regard to the ability to induce proliferative responses and LAK cell activity in bovine peripheral blood mononuclear cells in vitro. In both instances, chimerization had no effect on IL-2 activity. Similarly, the LKT component was unaffected in its ability to induce an effective immune response after immunization. The adjuvant properties of IL-2 have been established in a number of models, and this effect was tested by using the chimera. A multiple-injection protocol of LKT-IL-2 was compared with single-dose administration of LKT. The results obtained indicate that while there was no increase in specific antibody production, the IL-2 component of the chimera may be able to affect antigen-specific proliferation, as assessed by limiting-dilution analysis. Use of cytokine-antigen chimeras may provide a valuable antigen-adjuvant formulation that is simple to produce and purify and thus have economic advantages over conventional preparations. Furthermore, chimerization will also ensure that the adjuvant acts at the same site as the antigen, thus optimizing immunostimulatory activity.

Animals

Ocular integrity after refractive surgery: effects of photorefractive keratectomy, phototherapeutic keratectomy, and radial keratotomy.

The effects on ocular integrity of three surgical techniques to correct refractive errors were compared. Ten matched pairs of eyes (right and left eyes) freshly enucleated from pigs underwent radial keratotomy (RK) (four incisions, 3.50-millimeter clear zone) in one eye and photorefractive keratectomy (PRK) with the 193-nanometer excimer laser (10.00- diopter myopic correction, 6.00-millimeter ablation diameter) in the fellow eye. Another 16 eyes under- went phototherapeutic keratectomy (PTK) at various depths; another 10 eyes were used as controls. All eyes were subjected to lateral compression by a bench press, with gradually increasing pressure applied until the globe ruptured. The rupture site was photographed. Of the 10 control eyes, 9 ruptured at the sclera and 1 ruptured at the optic nerve. All eyes that had undergone RK ruptured at the sites of the corneal incisions, whereas the eyes that had PRK ruptured at the sclera (P < .01). After PTK, corneal rupture upon compression occurred when ablations were about 40% depth or greater. Corneas that have been subjected to PRK thus appear less predisposed to rupture due to blunt trauma than do eyes that have undergone RK.

Animals

Corneal surface after deepithelialization using a sharp and a dull instrument.

Manual removal of the corneal epithelium currently precedes excimer laser photorefractive keratectomy. To assess the smoothness of the corneal surface following this procedure, six paired human eye-bank eyes (12 eyes) were manually deepithelialized using a blunt instrument (Paton spatula) on one eye and a sharp instrument (surgical blade) on the contralateral eye; all deepithelializations were done by the same surgeon. The 12 corneas were then resected and processed in an identical fashion for scanning electron microscopy. The microphotographs obtained showed that among the corneas deepithelialized with the blunt spatula, two had a smooth surface, and four had variable amounts of residual epithelial cells and basement membrane, confirmed by light microscopy. The surface of five corneas deepithelialized with the sharp instrument were slightly rougher than the former, with occasional linear scratches; one cornea retained a small amount of basement membrane. Removal of epithelium appears to be more complete when a sharp instrument is used. These findings may have clinical relevance, since residual epithelium and basement membrane after deepithelialization may influence the depth of ablation subsequently achieved with the excimer laser.

Basement Membrane

Cell-mediated cytotoxic responses in lungs following a primary bovine herpes virus type 1 infection.

Non-major histocompatibility complex (MHC) restricted cytotoxicity is an important part of the immune reaction mounted in response to bovine herpes virus type 1 (BHV-1) infection. In this study, we evaluated the effect of BHV-1 infection on the ability of lung parenchyma leucocytes (LPL), cranial tracheobronchial lymph node cells (BLNC) and peripheral blood mononuclear leucocytes (PBML) to mediate this function. While LPL from non-infected calves mediated cytotoxicity against BHV-1-infected cells, a similar activity could not be detected in PBML or BLNC. In contrast, both LPL and PBML from naive calves could mediate cytotoxicity against K562 target cells but only after activation with interleukin-2 (IL-2). BLNC were unable to kill K562 cells. Infection of calves with BHV-1 enhanced the ability of LPL and PBML to kill BHV-1-infected cells. This enhancement was detected as early as Day 1 after infection in LPL whereas it could only be detected in PBML 8 days after infection. The results demonstrate that the leucocyte population present at the site of infection was able to mediate a potentially important antiviral function and that this function was enhanced rapidly in response to infection. Thus LPL-mediated cytotoxicity may be an important mechanism for the recovery from BHV-1 infection.

Animals

Clinical and immunological effects of single bolus administration of recombinant interleukin-2 in cattle.

Recombinant bovine interleukin-2 (rBoIL-2) was administered as a single intramuscular bolus to healthy calves to determine the minimal dose capable of exerting a biological response. Doses ranging from 2.5 to 0.05 micrograms rBoIL-2/kg did not induce pyrexia, diarrhea, or depression, nor did they alter any blood chemistry or hematological parameters commonly associated with IL-2 toxicity. Moreover, the only significant immunological change observed was a reduction in the number of peripheral blood lymphocytes identified with the monoclonal antibodies B7A, BAQ4A (WC1+ cells), CACTB6A (WC2+ cells) and DH59B (monocytes). The decrease in cells associated with these markers did not influence non-MHC restricted cytotoxicity or in vitro lymphocyte proliferative responses to mitogens and IL-2. The treatments had no effect on delayed type hypersensitivity responses to phytohemagglutinin. These results indicate that IL-2 may be involved in the regulation of trafficking patterns of a unique subpopulation of lymphocytes in cattle.

Animals