Biomedical subjects

C Maldonado

Publications and source records attributed to C Maldonado.

At least 37 records · Page 2Linked to original sources

PubMed2000

Electrically stimulated detrusor myoplasty.

PURPOSE: Many children with spina bifida and other causes of neurogenic bladder rely on clean intermittent catheterization to empty the hyporeflexic or areflexic bladder. Direct bladder and sacral nerve root stimulation have been met with limited success. We studied the electrical stimulation of a rectus abdominis muscle flap wrapped around the bladder to achieve bladder contractility and emptying. MATERIALS AND METHODS: The feasibility of performing rectus detrusor myoplasty in humans was first studied in 8 cadavers. In male and female cadavers it was possible to wrap the distended bladder completely with the rectus abdominis muscle. The rectus abdominis muscle was surgically dissected with preservation of its insertion on the pubis bone and rotation of its mid section behind the bladder to effect a complete bladder wrap. The deep inferior epigastric artery and veins, and 2 most caudal intercostal nerves were preserved. This unilateral rectus abdominis muscle flap was then electrically stimulated with 2 pairs of bipolar electrodes inserted into the muscle near the nerve entrance. Stimulation frequencies of 40, 60 and 80 Hz. were used in each of the 8 dogs. The increase in intravesical pressure over baseline, compliance and post-void residual were measured. Paired Student's t tests were used for statistical comparisons. RESULTS: The increase in intravesical pressure ranged 35 +/- 5 to 45 +/- 7 cm. H2O at stimulation frequency 40 and 80 Hz., respectively. Post-void residual was 27 +/- 4%, 22 +/- 3% and 26 +/- 3% at stimulation frequencies 40, 60 and 80 Hz., respectively. Intravesical pressure was significantly increased over baseline bladder pressure (p <0.05). CONCLUSIONS: Electrically stimulated detrusor myoplasty results in uniform increases in intravesical pressure and reasonable bladder emptying in an animal model. We are currently investigating detrusor myoplasty in a chronic study to determine whether it can be used for enhanced bladder emptying in children with poor detrusor contractility.

Abdominal Muscles↗

PubMed1999

Scoring of skin rejection in a swine composite tissue allograft model.

BACKGROUND: For the first time, we define and correlate visual and histologic grading systems of composite tissue allograft (CTA) skin rejection in a large-animal model and determine the utility of these grading systems for early diagnosis and monitoring of rejection. MATERIALS AND METHODS: Sixteen pairs of outbred swine underwent transplant of a forelimb osteomyocutaneous free flap. Group I (n = 6) did not receive immunosuppressive therapy. Group II (n = 10) received oral cyclosporin A, mycophenolate mofetil, and prednisone. The flap was visually inspected and protocol skin biopsies were taken at frequent intervals over a 90-day period. Visual Grades 0 (no rejection) to 4 (severe rejection) were assigned based on skin color, bleeding from biopsy site, and blister formation. Histologic Grades 0 to 4 were assigned based on the degree of vasculitis, folliculitis, dermal inflammation, and epidermal degeneration present. RESULTS: All Group I animals progressively rejected their graft by Day 7. Group II grafts survived from 19 and 90 days; 93% of 115 biopsy specimens were read to be within +/-1 histologic score of their assigned flap visual grade. Visual assessment carried an 8% false positive and 39% false negative rate with regard to biopsy-proven rejection. However, 81% of missed rejection specimens were histologic Grade 1. Biopsy, when visually indicated, would detect all rejection episodes when histologically Grade 1 or 2 and still potentially reversible. CONCLUSIONS: Visual scoring of CTA skin serves as a useful tool for initially detecting rejection, but repeated histologic evaluation is necessary for monitoring the subsequent course of the graft.

Animals↗

PubMed1999

Pharmacokinetic advantage of intra-arterial cyclosporin A delivery to vascularly isolated rabbit forelimb. I. Model development.

Effective antirejection therapy with minimal systemic morbidity is required if limb transplantation is to become a clinical reality. We investigated whether i.a. infusion of cyclosporin A (CSA) into the vascularly isolated rabbit forelimb will distribute drug homogeneously to the tissues and produce higher local drug levels than same-dose i.v. treatment, thereby improving the therapeutic index. CSA 4.0 mg/kg/day was infused continuously via osmotic minipump into either the right brachial artery (i.a. group) or jugular vein (i.v. group) of New Zealand rabbits. Ligation of all muscles at the right mid-arm level was performed in the i.a. group to eliminate collateral circulation and simulate allografting, while leaving bone and neurovasculature intact. On day 6, CSA concentrations were measured in skin, muscle, bone, and bone marrow samples taken from different compartments of the right and left forearms in the i.a. group and right forearm only in the i.v. group. There were no significant differences between compartmental CSA levels in all tissues examined on the locally treated, right side during i.a. infusion, indicating that drug streaming from the catheter tip is not occurring in our model. During i.a. infusion, mean CSA concentrations were 4- to 7-fold higher in the right limb than in the left limb in all four tissues examined. Tissue CSA levels in the left limb were equivalent to those achieved during i.v. infusion, but CSA concentrations in blood, kidney, and liver were higher during i.a. infusion. These favorable, preliminary, single-dose pharmacokinetic results warrant further investigation in our novel rabbit model.

Animals↗

PubMed1999

Pharmacokinetic advantage of intra-arterial cyclosporin A delivery to vascularly isolated rabbit forelimb. II. Dose dependence.

A vascularly isolated rabbit forelimb model simulating conditions of composite tissue allografting was used to determine the regional pharmacokinetic advantage achievable in extremity tissue components during i.a. cyclosporin A (CSA) administration. CSA was infused continuously via osmotic minipump into the right brachial artery of New Zealand rabbits at multiple doses ranging from 1.0 to 8.0 mg/kg/day. On day 6, CSA concentrations were measured in aortic whole blood, as well as in skin, muscle, bone, and bone marrow samples from both right and left forelimbs. The variation of right-sided mean CSA concentrations with dose was tissue dependent and saturable in the case of skin and bone, whereas left-sided tissue concentrations correlated significantly with systemic blood levels. At 1.0 mg/kg/day, there were no significant differences between right and left mean CSA concentrations for all four tissues examined. However, with a doubling of the i.a. dose, huge increases in local tissue CSA concentrations were produced with only very modest increases in systemic whole-blood and tissue drug levels, resulting in a 4-fold regional advantage (right/left ratio of CSA concentrations) in bone and bone marrow, 7-fold in muscle, and 14-fold in skin. With further dose increases to 8.0 mg/kg/day, the regional advantage decreased to 4-fold in skin, increased to 9-fold in bone marrow, remained relatively constant in bone, and initially decreased and then increased to 9-fold in muscle. These favorable pharmacokinetic results suggest that reduced, local doses of CSA might be useful in preventing extremity composite tissue allograft rejection with decreased systemic drug exposure.

Animals↗

PubMed1999

Pharmacokinetics of intra-arterial delivery of tacrolimus to vascularly isolated rabbit forelimb.

A vascularly isolated rabbit forelimb model simulating conditions of composite tissue allografting was used to determine the regional pharmacokinetic advantage achievable in extremity tissue components during i.a. tacrolimus (FK506) administration. FK506 was infused continuously via osmotic minipump into the right brachial artery of New Zealand rabbits at 0.05, 0.1, and 0.2 mg/kg/day. On day 6, FK506 concentrations were measured in aortic whole blood, heart, lung, liver, kidney, spleen, and fat, as well as in skin, muscle, bone, and bone marrow samples from both right and left forelimbs. The relative tissue concentrations of FK506 in descending order were [spleen approximately lung approximately kidney] > [heart approximately skin approximately muscle] > [fat approximately bone marrow] > [liver approximately bone approximately blood]. In marked contrast to previous results with i.a. cyclosporin A infusion, only a minimal regional advantage of local FK506 delivery (mean right/left concentration ratios 1.0-1.4) was obtained in all forearm tissues over the dose range studied. For each limb tissue, left-sided FK506 concentrations significantly correlated with systemic blood levels, and the left-sided tissue-to-whole-blood concentration ratio did not vary significantly with dose. We conclude that FK506 is pharmacokinetically inferior to cyclosporin A for continuous i.a. administration to the vascularly isolated rabbit forelimb, and hypothesize that this difference is the result of differences in the distribution of each drug within whole blood. Our findings suggest that, despite its demonstrated efficacy in experimental and clinical transplantation, FK506 would not be an appropriate immunosuppressant to deliver via the i.a. route for prevention of limb allograft rejection.

Animals↗

PubMed1999

Long-term survival of an extremity composite tissue allograft with FK506-mycophenolate mofetil therapy.

BACKGROUND: High-dose tacrolimus (FK506) monotherapy has significantly prolonged rat hindlimb allograft survival. With an eye toward direct clinical application, we used a large-animal extremity composite tissue allograft model to assess the antirejection efficacy and systemic toxicity of combination FK506-mycophenolate mofetil (MMF) treatment. METHODS: Radial forelimb osteomyocutaneous flap transplants were performed between size-matched outbred pigs assigned to one of two groups: 5 control pigs received no immunosuppression and 9 animals received a once-daily oral FK506-MMF-prednisone regimen. Rejection was assessed by visual inspection of flap skin and was correlated with serial histopathologic examination of skin biopsy specimens. RESULTS: In all control pigs the flap was completely rejected on day 7. Of the 9 pigs receiving treatment, 3 died from pneumonia on days 29, 30, and 83 without signs of rejection and another died from gastric rupture on day 42 with persistent mild rejection. The remaining 5 animals were free of rejection at the end of the 90-day follow-up period (P < 0.005 vs controls). Overall, 5 pigs had pneumonia, 4 septic arthritis, 3 toe abscesses, and 5 diarrhea and decreased weight gain. CONCLUSIONS: Combination oral FK506-MMF treatment provided a superior antirejection effect but more produced more toxicity than that previously demonstrated with cyclosporin A-MMF therapy in our model. Our results suggest that reduction of FK506 or MMF doses might decrease both infectious and drug-specific side effects while still providing adequate prophylaxis against rejection.

Animals↗

PubMed1998

Long-term composite tissue allograft survival in a porcine model with cyclosporine/mycophenolate mofetil therapy.

BACKGROUND: Low-dose cyclosporine (CsA)/mycophenolate mofetil (MMF) therapy has significantly reduced the frequency of rejection and drug-induced side effects in rat hindlimb allograft recipients. With an eye toward direct clinical application, we developed a large-animal extremity composite tissue allograft model to assess the antirejection efficacy and systemic toxicity of combination CsA/MMF treatment. METHODS: Radial forelimb osteomyocutaneous flap transplants were performed between size-matched, outbred pigs assigned to one of two groups: 5 control pigs received no immunosuppression, and 10 pigs received a once-daily oral CsA/MMF/prednisone regimen. Rejection was assessed by visual inspection of flap skin and correlated with serial histopathologic examination of skin biopsies. RESULTS: In all control pigs, the flap was completely rejected on day 7. Of the 10 pigs receiving treatment, one died from pneumonia and an another from an anesthetic complication on days 19 and 30, respectively, without signs of rejection. Two flaps were lost on days 25 and 29 from severe rejection. Three pigs were free of rejection at the end of the 90-day follow-up period, and three had stable mild-to-moderate rejection at 90 days (P= 0.0007 vs. controls). White blood cell and platelet counts, serum creatinine values, and liver function tests remained normal in all animals receiving immunosuppressive therapy. CONCLUSIONS: Our results, to our knowledge, demonstrate for the first time that rejection can be significantly delayed in a large-animal composite tissue allograft model including skin using only orally administered agents dosed according to clinically relevant strategies without significant drug-specific systemic side effects.

Animals↗

PubMed1998

Quantification of the morphological features of a full microvascular network.

Investigations into the changes that occur in microvasculature following the surgical procedure called delay have brought about the need for a computer system capable of quantifying the morphological features of a full microvascular network in terms of average vessel length, diameter, and tortuosity. Both the formulaic conventions that have been developed to measure these quantities as well as their implementation in the form of a HP-9000/UNIX based computer software system that we developed specifically for this purpose are discussed. Reliability studies performed using the final system to measure the microcirculatory network of a mouse latissmus dorsi muscle (LDM) showed 95% confidence intervals within 5% of means and coefficients of variability within 7% of means for all quantities measured in large (150-300 microns), medium (50-150 microns), and small (< 50 microns) diameter vessels. These variations were significantly smaller than the changes that were observed in a preliminary study comparing these microvascular network parameters before and after delay in the hairless mouse LDM, showing the proposed quantification methods to be well suited to the study of the microvascular changes following delay. It is hoped that the formulaic conventions, implementation process and reliability data will provide a useful comparison for other researchers interested in measuring similar features of microcirculatory networks.

Animals↗

PubMed1997

Diversity of pituitary cells in primary cell culture. An immunocytochemical study.

In cell cultures of dispersed rat anterior pituitary, the specific identification of each cell type based on their staining properties and the ultrastructural features of secretory granules has proved to be unreliable. The existence of pituitary cell subtypes and the striking remodeling of the cell surface and intracellular organelles, further complicate the specific identification of pituitary cell populations. An immunocytochemical study of dissociated pituitary cells in culture was carried out to identify the cellular hormonal content by applying specific antibodies against prolactin (PRL), and growth (GH), luteinizing (LH beta), adrenocorticotrophic (ACTH) and thyrotrophic (TSH) hormones. Specifically bound IgG was exposed by the electron microscope with protein A-gold complex. Typical lactotrophs, somatotrophs and gonadotrophs are easily recognized because they retain the main features described in the pituitary tissue in situ. Other undefined groups of cells bearing small or medium round secretory granules can be identified by immunocytochemistry as PRL, GH or TSH producing cells. The latter technique was critical for the characterization of the hormonal content of secretory granules, the shape, size, electron density and cytoplasmic distribution of which differ substantially from those described in the intact gland. Cells displaying rare small oval or sharp pointed secretory granules were identified as gonadotrophs with anti-LH beta, while corticotrophs showed granules with irregular profiles not previously reported in the gland. These remarkable morphological changes appear to be related to the interruption of the flow of hypothalamic hormones and the disruption of structural and paracrine interrelationships. This investigation reveals that immunocytochemistry is essential for the specific recognition of the various pituitary cell types, and particularly of atypical cells exhibiting morphological features not found in the pituitary gland in situ.

Adrenocorticotropic Hormone↗

PubMed1997

Role of adenosine receptors in late preconditioning against myocardial stunning in conscious rabbits.

Conscious rabbits underwent six 4-min coronary occlusions interspersed with 4-min periods of reperfusion for 2 consecutive days (days 1 and 2 of stage I); 2 wk later, they underwent the same protocol (days 1 and 2 of stage II) except that they received either 8-(p-sulfophenyl)theophylline (SPT) on day 1 (group I, n = 5) or 2-chloro-N6-cyclopentyl-adenosine (CCPA) on the day before day 1 (group II, n = 6). In both groups I and II, on day 1 of stage I, systolic wall thickening (WTh) remained significantly depressed for several hours, indicating myocardial stunning; on day 2, however, the total deficit of WTh was approximately 50% less than on day 1 (P < 0.01), indicating the development of late preconditioning (PC) against myocardial stunning. Despite administration of SPT, in group I the deficit of WTh during stage II was 55% less on day 2 than on day 1 (P < 0.05). Similar results were obtained in three other rabbits treated with PD-115199 on day 1. In group II, pretreatment with CCPA during stage II failed to decrease the deficit of WTh on day 1. This study presents a new conscious rabbit model for studying myocardial stunning that is relatively inexpensive and technically less demanding than larger animal models. In this model, the development of late PC against myocardial stunning is not blocked by nonselective blockade of adenosine receptors with either SPT or PD-115199, nor is it induced by activation of adenosine A1 receptors with CCPA, indicating that adenosine receptors are not involved in the pathogenesis of this phenomenon.

Animals↗

PubMed1997

Selective activation of A3 adenosine receptors with N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide protects against myocardial stunning and infarction without hemodynamic changes in conscious rabbits.

To examine the cardioprotective role of A3 adenosine receptors during myocardial ischemia/reperfusion injury, we tested the effect of N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA), a potent and selective A3 adenosine receptor agonist, in models of myocardial stunning and infarction in chronically instrumented conscious rabbits. In phase I (studies of myocardial stunning), rabbits were subjected to six 4-minute coronary occlusions, each separated by 4-minute reperfusion periods, after which the recovery of systolic wall thickening was measured (ultrasonic crystals). In phase II (studies of myocardial infarction), rabbits were subjected to a 30-minute coronary occlusion followed by 3 days of reperfusion. In both phases, IB-MECA was administered as an intravenous bolus (100 micrograms/kg) 10 minutes before the first coronary occlusion. This dose of IB-MECA was determined in pilot studies to have no effect on heart rate, arterial blood pressure, or plasma histamine concentration in rabbits. In phase I, IB-MECA markedly improved the recovery of wall thickening after the six occlusion/reperfusion cycles, and this effect was sustained throughout the 5-hour observation period; the total deficit of wall thickening (a measure of the overall severity of myocardial stunning) was reduced by 68% (control, 129 +/- 16 arbitrary units, n = 7; IB-MECA, 41 +/- 6 arbitrary units, n = 6; P < .01). The protective effects of IB-MECA against stunning were completely blocked by pretreatment with the nonselective adenosine receptor antagonist 8-p-sulfophenyl theophylline or the specific protein kinase C inhibitor chelerythrine. In phase II, IB-MECA reduced myocardial infarct size by 61%; infarct size (tetrazolium staining) was 41 +/- 4% of the risk region in control animals (n = 8) and 16 +/- 6% in IB-MECA-treated animals (n = 8, P < .01). These results demonstrate that in conscious rabbits the A3 adenosine receptor agonist IB-MECA confers a powerful protection against both reversible (stunning) and irreversible (infarction) injury during acute myocardial ischemia and reperfusion by a protein kinase C-mediated pathway, suggesting that selective activation of A3 receptors is an effective means of protecting the ischemic myocardium without hemodynamic changes.

Adenosine↗

PubMed1996

Attenuation of vasospasm and capillary no-reflow by ischemic preconditioning in skeletal muscle.

Vasospasm and capillary no-reflow are common complications following replantation and free flap transfer. The purpose of the present study was to clarify whether vasospasm and capillary no-reflow which are induced by prolonged warm ischemia/reperfusion can be attenuated by ischemic preconditioning in the vascular isolated cremaster muscle model. Male Sprague-Dawley rats were anesthetized with pentobarbital. Arteriole diameter and capillary perfusion were measured utilizing intravital microscopy. In the control group, the cremasters sustained 4-hour warm global ischemia followed by 60-minute reperfusion. In the ischemic preconditioning group, the cremasters were subjected to one cycle of 45-minute ischemia followed by 15-minute reperfusion prior to 4-hour warm global ischemia followed by 60-minute reperfusion. The results from this experiment showed that ischemic preconditioning significantly attenuated ischemia/reperfusion-induced vasospasm and capillary no-reflow which occur early during reperfusion after prolonged warm ischemia in skeletal muscle. The mechanism of this phenomenon remains to be elucidated.

Animals↗

PubMed1996

Mechanisms of triggered activity induction at the border zone of normal and abnormal cardiac tissue.

Ionic mechanisms that may be involved in inducing triggered activations at the border zone (BZTAs) of normal and abnormal Purkinje fiber segments were investigated. In a two-chamber bath, fibers were divided into a normal segment and segment treated with ethylenediaminetetraacetic acid to stimulate electrophysiologic alterations 24 hours after infarct. Interventions to normal segments included 1.8 mM lidocaine (n = 10), 3 x 10(-4) mM tetrodotoxin (n = 5), 10(-3) mM aconitine (n = 4), 3 mM cesium chloride (n = 7), 10(-2) mM verapamil (n = 4), and 6-8 mM (n = 7) of K+. Ethylenediaminetetraacetic acid (3.3 mM) prolonged action potentials and induced low diastolic potentials in the normal segment border zone. Tetrodotoxin, lidocaine, and high K+ levels suppressed BZTAs; cesium chloride and aconitine increased BZTAs; and verapamil did not reduce BZTAs. The finding that BZTAs were not abolished by verapamil suggests that abnormal automaticity is not a mechanism. Apparently, BZTAs depend on the Na+ inward current activated by depolarization of the membrane secondary to depolarization of adjacent cells.

Action Potentials↗

PubMed1996

Caparratriene, an active sesquiterpene hydrocarbon from Ocotea caparrapi.

Caparratriene (1), a new sesquiterpene hydrocarbon with significant growth inhibitory activity (IC50 = 3.0 +/- 0.5 x 10(-6) M) against CEM leukemia cells, was isolated from the oil of Ocotea caparrapi (Nates) Dugand. The structure of 1, determined by spectroscopic techniques, corresponded to (E,E)-3,7,11-trimethyl-2,4,10-dodecatriene (C15H26).

Antineoplastic Agents, Phytogenic↗

PubMed1996

Cellular and subcellular localization of glycogenin in chicken retina.

We investigated the cellular and subcellular localization of glycogenin in chicken retina using a polyclonal antibody raised against chicken muscle glycogenin. The antiserum recognized both free and glycogen-bound glycogenin on dot blots. Immunocytochemistry revealed an uniform staining of all the retina layers except for the outer segments and ganglion cells layers that were very weakly stained and for the inner segments layer and a zone between the inner nuclear and inner plexiform layers that were heavily stained. Electron microscopy of neuronal cells showed immunoreactivity localized in the cytoplasm and nucleus. This is the first description of the cellular and subcellular localization of glycogenin. Our results suggest that the biosynthesis of glycogen could begin in both, the cytoplasm and nucleus of the neurone.

Animals↗

PubMed1996

Neuroradiology and clinical aspects of Usher syndrome.

We describe the neurological evaluation and MRI analysis of 30 patients, belonging to 16 families with Usher syndrome (US) type I and type II (US1 and US2). In addition to the classic visual and audiological abnormalities seen in these patients, we observed abnormal gait in 88.9% of US1 and in 66.7% of US2 patients and abnormal coordination in 33.4% of US1, and in 58.3% of US2. Borderline mental retardation, depression or bipolar affective disorder were observed in 16.7% of US1 and 33.3% of US2 patients. MRI analysis showed cerebellar abnormalities in 50% of US1 and 75% of US2 patients, but no clear correlation was observed between structural abnormalities and clinical findings. A pattern for the MRI classification of US patients is suggested.

Abnormalities, Multiple↗

PubMed1996

Exogenous Fibronectin to Prevent Neointimal Hyperplasia after Balloon Angioplasty.

This study examined the effect of fibronectin to prevent restenosis in a microswine model after balloon angioplasty of the right iliac artery. Immediately following angioplasty, fourteen hypercholesterolemic microswines were randomized to receive fibronectin (223.5 mg, n = 8) or saline containing albumin (75.6 mg, n = 6). At 60-days post-angioplasty, the angioplasty-injured and intact contralateral arteries were examined with angiography and histopathology. With angiography, there was no significant difference in the luminal diameters of angioplasty arteries compared with the intact contralateral vessels. Histological examination of angioplasty-injured vessels showed neointimal hyperplasia. The intimal areas of angioplasty-injured vessels, in the placebo and fibronectin groups, were much larger than the areas of their contralateral vessels (fibronectin 287 +/- 160 vs. 138 +/- 88 µm2, p = 0.018; and the placebo (1,245 +/- 1,567 vs. 248 +/- 219 µm2, p = 0.041). Mean total cholesterol levels of both groups were maintained at levels > 400 mg/dl throughout the study period. CONCLUSION: At sixty days after balloon angioplasty injury: 1) fibronectin did not only not prevent neointimal hyperplasia, in some animals, it increased neointimal growth; and 2) angiographic results were not sensitive enough to quantify changes observed in histologic findings.

Journal Article↗

PubMed1996

Brevetoxin-3 (PbTx-3) on mouse liver slices: a histological study.

Brevetoxin-3 (PbTx-3) is a marine toxin produced by the dinoflagellate Ptychodiscus brevis. Its effects on excitable tissues have been the main subject of studies, but little is known about how it affects non-excitable tissues. To study possible non-neural effects of PbTx-3 (78nM), its effects on hepatic cell structure in vitro were evaluated. PbTx-3 caused hypertrophy and increased vacuolation of hepatocytes, and an increase in basophilia in the perivenous area of the lobules. Ultrastructurally, it was evident that the vacuolation was related to swelling of the endoplasmic reticulum, changes that probably account for the increased basophilic reaction of the cells. The swelling in smooth endoplasmic reticulum, degranulation of rough endoplasmic reticulum, the deformities and lytic cristae in the mitochondria, and the presence of active lysosomes are evidence of the PbTx-3 effects upon liver cells. These responses are probably caused by the liver's detoxification role on the PbTx-3.

Animals↗