Static Magnetic Fields Modulate the Response of Different Oxidative Stress Markers in a Restraint Stress Model Animal.

Stress is a state of vulnerable homeostasis that alters the physiological and behavioral responses. Stress induces oxidative damage in several organs including the brain, liver, kidney, stomach, and heart. Preliminary findings suggested that the magnetic stimulation could accelerate the healing processes and has been an effective complementary therapy in different pathologies. However, the mechanism of action of static magnetic fields (SMFs) is not well understood. In this study, we demonstrated the effects of static magnetic fields (0.8 mT) in a restraint stressed animal model, focusing on changes in different markers of oxidative damage. A significant increase in the plasma levels of nitric oxide (NO), malondialdehyde (MDA), and advanced oxidation protein products (AOPP), and a decrease in superoxide dismutase (SOD), glutathione (GSH), and glycation end products (AGEs) were observed in restraint stress model. Exposure to SMFs over 5 days (30, 60, and 240 min/day) caused a decrease in the NO, MDA, AGEs, and AOPP levels; in contrast, the SOD and GSH levels increased. The response to SMFs was time-dependent. Thus, we proposed that exposure to weak-intensity SMFs could offer a complementary therapy by attenuating oxidative stress. Our results provided a new perspective in health studies, particularly in the context of oxidative stress.

Novel GP64 envelope variants for improved delivery to human airway epithelial cells.

Lentiviral vectors pseudotyped with the baculovirus envelope protein GP64 transduce primary cultures of human airway epithelia (HAE) at their apical surface. Our goal in this study was to harness a directed evolution approach to develop a novel envelope glycoprotein with increased transduction properties for HAE. Using error-prone PCR, a library of GP64 mutants was generated and used to prepare a diverse pool of lentiviral virions pseudotyped with GP64 variants. The library was serially passaged on HAE and three GP64 mutations were recovered. Single-, double- and the triple-combination mutant envelope glycoproteins were compared with wild-type GP64 for their ability to transduce HAE. Our results suggest that lentiviral vectors pseudotyped with evolved GP64 transduced HAE with greater efficiency than wild-type GP64. This effect was not observed in primary cultures of porcine airway epithelial cells, suggesting that the directed evolution protocol was species specific. In summary, our studies indicate that serial passage of a GP64 mutant library yielded specific variants with improved HAE cell tropism, yielding tools with the potential to improve the success of gene therapy for airway diseases.

Phosphodiesterase-4 inhibition improves corticosteroid insensitivity in pulmonary endothelial cells under oxidative stress.

BACKGROUND: Several clinical studies have shown that smoking in asthmatics and chronic obstructive pulmonary disease patients is closely associated with corticosteroid refractoriness. In this work, we have analyzed glucocorticoid insensitivity in human pulmonary artery endothelial cells (HPAECs) under cigarette smoke extract (CSE) exposure as well as the possible additive effects of the combination therapy with a phosphodiesterase (PDE)-4 inhibitor. METHODS: Interleukin (IL)-8 was measured in cell supernatants by ELISA. Histone deacetylase (HDAC), histone acetylase (HAT), and intracellular cAMP levels were measured by colorimetric assays and enzyme immunoassay, respectively. PDE4 isotypes and glucocorticoid receptor (GR)-α and ß expression were measured by real-time RT-PCR. RESULTS: The PDE4 inhibitor rolipram dose dependently inhibited the IL-8 secretion induced by CSE 5%. In contrast, dexamethasone 1 µM did not show inhibitory effect on IL-8 secretion. Combination of subeffective rolipram concentrations at 10 nM increased the inhibitory effect of dexamethasone to ~45% of inhibition. Cigarette smoke extract 5% inhibited HDAC activity and increased HAT activity generating glucocorticoid insensitivity. Rolipram did not modify the HDAC activity, however partially inhibited the increase in HAT activity at 1 µM. PDE4 isotypes were up-regulated by CSE 5% with the consequent cAMP down-regulation. Dexamethasone reduced all PDE4 isotypes expression and showed additive effects with rolipram enhancing cAMP levels. Furthermore, rolipram enhanced GR-α expression and inhibited the increase in GR-ß induced by CSE. CONCLUSIONS: Combination of rolipram and dexamethasone shows additive properties in HPAECs under glucocorticoid insensitive conditions. These results may be of potential value in future anti-inflammatory therapies using combination of PDE4 inhibitors and glucocorticoids.

Bosentan inhibits cigarette smoke-induced endothelin receptor expression in pulmonary arteries.

The endothelin (ET) system contributes to lung vascular tension and remodelling in smokers and chronic obstructive pulmonary disease (COPD) patients. This study examined the effect of cigarette smoke (CS) on ET receptor A (ET(A)) and B (ET(B)) expression in human pulmonary artery smooth muscle cells (HPASMCs) and human small intrapulmonary arteries, as well as their functional consequences. CS extract (CSE) increased ET(A) and ET(B) expression in HPASMCs and small intrapulmonary arteries, which was attenuated by bosentan, the ET(A) antagonist BQ123 and the ET(B) antagonist BQ788, and by blocking ET-1 with a monoclonal antibody against ET-1, suggesting a feed-forward mechanism mediated by ET-1 release. ET receptor (ETR) antagonism attenuated the CSE-induced HPASMC proliferation. Furthermore, CSE exposure increased the acute ET-1-induced small intrapulmonary artery contraction, which was attenuated by bosentan, BQ123 and BQ788. Pulmonary arteries from smokers and COPD patients showed a higher expression of ET(A) and ET(B) than those of nonsmoker patients. These results show a novel mechanism by which ETR blockade attenuates CS-induced ETR overexpression and, subsequently, small intrapulmonary artery tension. These data may be of potential value to explain therapeutic effects of bosentan in some forms of disproportionate pulmonary hypertension in COPD patients.

Cigarette smoke exposure up-regulates endothelin receptor B in human pulmonary artery endothelial cells: molecular and functional consequences.

BACKGROUND AND PURPOSE: Pulmonary arteries from smokers and chronic obstructive pulmonary disease patients show abnormal endothelium-dependent vascular reactivity. We studied the effect of cigarette smoke extract (CSE) on endothelin receptor B (ET(B) ) expression in human pulmonary artery endothelial cells (HPAECs) and its role in endothelial dysfunction. EXPERIMENTAL APPROACH: ET(B) receptor expression was measured by real time RT-PCR, Western blot and immunofluorescence. Cell contraction, intracellular Ca(2+) , F/G-actin, RhoA activity, myosin light chain phosphorylation, ET, NO, thromboxane (Tx)A(2) and reactive oxygen species (ROS) were measured by traction microscopy, fluorescence microscopy, phalloidin fluorescence, colorimetric assay, Western blot, elisa and DCFDA fluorescence respectively. KEY RESULTS: Cigarette smoke extract dose-dependently increased ET(B) receptor expression in HPAECs after 24h incubation. CSE-induced ET(B) expression was attenuated by bosentan, the ET(B) receptor antagonist BQ788, the Rho kinase antagonist Y27632 and the antioxidant N-acetylcysteine. A monoclonal antibody to ET-1 prevented CSE-induced ET(B) receptor overexpression. Twenty-four hour exposure to ET-1 dose-dependently increased ET(B) receptor expression, mimicking the effect of CSE. CSE-induced ET(B) receptor overexpression caused greater cell contraction; increased intracellular Ca(2+) ; increased F/G-actin and RhoA activity; increased myosin light chain phosphorylation; augmented TxA(2) and ROS production; and decreased NO after acute ET-1 (10nM). These effects were attenuated by bosentan, BQ788, Y27632 and N-acetylcysteine. CONCLUSIONS AND IMPLICATION: Cigarette smoke extract induced ET(B) receptor overexpression by a feed forward mechanism mediated partly by ET release, promoting HPAEC dysfunction and attenuated by ET(B) receptor blockade, Rho kinase and ROS inhibition. These results provide support for the use of bosentan in CS-related endothelial dysfunction.

[Fibroepithelial polyp of the urethra in an adult]. / Pólipo fibroepitelial de uretra en un adulto.

We present the case of a fibroepithelial polyp of the prostatic urethra in a forty-five year old patient. This type of polyp is rare in the adult. The principal symptoms which the patient presented were haematuria and an intermittent reduction in micturitional flow. Endoscopic images of the tumour are shown and certain details of its histology; this is a polypoid lesion of loose connective tissue covered by typical urothelium, with occasional fascicles of smooth muscle, no prostatic-type glandular structures and no appreciable inflammatory component.

[Adrenal ectopy in epididymis]. / Ectopia adrenal en epidídimo.

OBJECTIVE: To report a case of ectopic adrenal in the epididymis. METHODS: A 17-year-old male consulted for a left varicocele and complaints referred to the right epididymis a year and a half after an episode of acute epididymitis. A nodule was excised from the epididymis. RESULTS: The pathological analysis showed an 0.4 cm nodule comprised of adrenocortical tissue. CONCLUSIONS: Ectopic adrenal cortical tissue is a benign lesion. Although routine search for this lesion is not indicated, it should be resected for histological analysis and differential diagnosis when it is found during a surgical procedure in this area.

[Distal ureteral lithiasis. ESWL versus ambulatory URS]. / Litiasis ureteral distal. LEOC versus URN ambulatoria.

OBJECTIVE: To present the results achieved in the treatment of 1802 distal ureteral stones treated at the Lithotripsy Unit of the San Cecilio University Hospital over the last 10 years (1990-2000). METHODS: Stones were treated by extracorporeal shock wave lithotripsy (ESWL) or ureteroscopy (URS). ESWL was the initial treatment in 81% of the cases (1460 calculi) and URS in the remaining 19% (342 stones). URS was performed for complication or failed ESWL (102 stones) and ESWL was performed for failed URS, basically due to stone migration (24 stones). Ureterolithotomy was required on 7 occasions. Sedation-analgesia with fentanyl and midazolam was routinely used in URS. Sedation was required in only 55% of the ESWL procedures. RESULTS: Elective ESWL resolved 93% of the cases, a percentage which is similar to that achieved with URS as first treatment. The ESWL retreatment rate was 1.3. URS was successful in 98% of the cases of failed ESWL. CONCLUSIONS: There are two treatment modalities for stones in the distal ureter: ESWL and URS. We advocate the use of outpatient URS with sedation preferably in the female patient, impacted stones, obstructive uropathy, stones larger than 2 cm and radiotransparent stones.

Histamine up-regulates phosphodiesterase 4 activity and reduces prostaglandin E2-inhibitory effects in human neutrophils.

OBJECTIVE: To investigate whether histamine produces up-regulation of phosphodiesterase (PDE) activity with functional consequences in human peripheral blood neutrophils. METHODS: PDE activity was studied by a radioisotopic method following anion-exchange chromatography. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for detection of mRNA transcripts of PDE4 subtypes. Cyclic AMP (cAMP) levels were measured by enzyme-immunoassay, and superoxide generation by cytochrome c reduction. TREATMENT: Neutrophils were incubated for 4 h with histamine (1 microM). RESULTS: PDE4 was the only isoenzyme activity increased in treated neutrophils. Kinetic analysis showed a approximately 1.5-fold increase in Vmax without alteration of Km values. cAMP content in treated cells was higher than resting values (0.52+/-0.07 vs. 2.75+/-0.31 pmol/10(6) cells). RT-PCR showed increased expression of mRNA transcripts for PDE4B in histamine-treated cells. Functionally, up-regulation of PDE4 reduced the inhibition by prostaglandin E2 of zymosan-induced superoxide generation. CONCLUSION: Histamine up-regulates PDE4 activity and produces heterologous desensitisation of human neutrophils.

[Renal cell carcinoma in patient with situs inversus and Kartagener syndrome]. / Carcinoma de células renales en paciente con situs inversus y síndrome de Kartagener.

A case of a adenocarcinoma renal in a patient with situs inversus complete to which was associated bronchiectasis and chronic sinusitis (kartagener's syndrome) is reported. It is the third case described in the international literature of a renal cells carcinoma in a patient with situs inversus totalis and the first in patient the one which has the triada classic of the kartagener's syndrome plus sterility. The tumor was discovered in a way incidental upon accomplishing a TAC toracoabdominal and was solved through nefrectomia for lumbotomy approach.

beta-Adrenoceptor stimulation up-regulates phosphodiesterase 4 activity and reduces prostaglandin E2-inhibitory effects in human neutrophils.

Human neutrophils were treated for 4 h with a combination of salbutamol (1 microM), a beta2-adrenoceptor agonist, and rolipram (30 microM), a selective phosphodiesterase 4 inhibitor, to investigate whether this treatment produces up-regulation of phosphodiesterase activity with functional consequences. Anion-exchange chromatography coupled with the use of selective activators and inhibitors demonstrated that a phosphodiesterase activity with characteristics of the isoenzyme type 4 was increased in drug-treated cells. Kinetic analysis showed a approximately 1.5-fold increase in Vmax without alteration of Km values. The augmented phosphodiesterase activity in drug-treated cells was abolished by actinomycin D. Cyclic AMP content in drug-treated cells was higher than resting values (27.28+/-2.79 pmol/10(6) cells vs. 0.34+/-0.03 pmol/10(6) cells). Reverse transcriptase-polymerase chain reaction showed increased expression of mRNA transcripts for PDE4B and PDE4A in drug-treated cells. Functionally, up-regulation of phosphodiesterase 4 reduced the inhibition by prostaglandin E2 of zymosan-induced superoxide generation.

[Non-functional renal paraganglioma associated with carotid paraganglioma and appendicular mucocele]. / Paraganglioma renal no funcionante asociado con un paraganglioma carotídeo y un mucocele apendicular.

OBJECTIVE: A case of nonfunctional renal paraganglioma associated with appendicular mucocele and metachronic carotid paraganglioma is presented. METHODS/RESULTS: A 68-year-old patient consulted for abdominal pain. Patient evaluation with US, CT and opague enema showed an elongated mass that compromised the cecum and a solid mass in the right kidney. The patient underwent appendectomy and radical right nephrectomy. The anatomopathological findings showed an appendicular mucocele and renal paraganglioma. The postoperative hormone analyses were normal. A carotid paraganglioma was removed 8 months later. CONCLUSIONS: Renal tumors arising from neural crest cells are uncommon. To our knowledge, this is the third case of nonfunctional renal paraganglioma reported in the literature. The associated appendicular mucocele was an incidental finding.

[Perioperative transfusion in prostatic surgery]. / Transfusión perioperatoria en la cirugía de próstata.

OBJECTIVE: To analyze the likelihood of perioperative transfusion in prostate surgery. METHODS: The records of patients who underwent prostate surgery at the Hospital Central de Asturias en 1996 were reviewed. The abstracted patient discharge records, encoded according to the ICD-9-CM, were analyzed for gender, age, diagnosis, surgical and non-surgical procedures, including transfusions of blood products. RESULTS: 29 (14.2%) of 204 operated patients required transfusion. By univariate analysis, the likelihood of requiring transfusion was significantly higher in patients with cancer (28.8%), patients undergoing radical prostatectomy (40%), and those undergoing more than one surgical procedure (21.2%). By multivariate logistic regression analysis, the surgical procedure, age and simultaneous urinary bladder surgery were related with the likelihood of requiring transfusion. The adjusted odds ratio for perioperative transfusion was 10.14 times in radical prostatectomy than in transurethral prostatectomy, 2.67 times for patients more than 75 years than in those aged less, and 2.85 times in patients undergoing simultaneous urinary bladder surgery than those who do not. CONCLUSIONS: Due to the wide range of variability of the adjusted likelihood for transfusion in prostate surgery (3.5% to 83.4%), estimation of the individual likelihood for transfusion could be useful for planning surgical transfusions, pretransfusion tests and autologous transfusion in these patients.

[Innervation of the human adrenal gland]. / Inervación de la glándula suprarrenal humana.

OBJECTIVE: To study the innervation of the human adrenal gland by immunohistochemical techniques using monoclonal antibodies against the 200-kDa phosphorylated neurofilament and S-100 protein, specific markers of the axons and Schwann cells, respectively. METHODS: 24 specimens of normal adrenal gland from patients that had undergone radical nephrectomy for a renal tumor were analyzed. The tissue was embedded in paraffin and prepared for analysis by immunohistochemical techniques with the indirect peroxidase method using primary monoclonal antibodies against the 200-kDA phosphorylated neurofilament and protein S-100. RESULTS: The nerves localized in the adrenal gland form a subcapsular plexus where branches arise that extend to the medulla and come into contact with the enterochromaffin cells and medullary neurons. The adrenal neurons are restricted to the medulla; they are dispersed or form ganglia. The satellite cells of the medullary ganglia and the sustentacular cells were positive for S-100. CONCLUSIONS: Most of the nerve fibers penetrate into the human adrenal gland, extending directly to the medulla apparently without coming into contact with the cells of the adrenal cortex and form similar connections to the synaptics with the chromaffin cells and the medullary neurons. The microganglia of the human adrenal gland are dispersed and comprised of few neurons.

[Efficacy of radiotherapy for upper urinary tract tumor following radical surgery]. / Eficacia de la radioterapia en los tumores del tracto urinario superior, tras cirugía radical.

OBJECTIVE: To determine the efficacy of postoperative irradiation for locoregional tumor recurrence and in the prevention of distant metastasis in patients with upper urinary tract tumor treated by radical surgery. METHODS: 51 urothelial tumors of the upper urinary tract, most of which had been treated by radical surgery (nephroureterectomy with resection of bladder cuff were analyzed. Since 1989 an adjuvant postoperative radiotherapy protocol (55 Gy) has been included for high grade and stage tumors. Tumor staging was done according to the Jewett-Marshall classification and the Mostoffi cytological grading. RESULTS: Of the total number of cases analyzed, 16 received postoperative irradiation; of these, 5 (31%) showed recurrence. The recurrence rate was 47% for those treated by radical surgery alone. In patients with stage C tumors, 5 out of 12 cases (41.6%) showed recurrence. The recurrence rate was higher (83%) for patients with stage C tumors treated by radical surgery alone. CONCLUSIONS: Adjuvant postoperative radiotherapy is advocated for high grade and stage upper urinary tract tumors. This therapeutic approach has been found to reduce the number of recurrences and, consequently, morbidity. Although the overall survival rate did not increase, a larger series may show more consistent results in this regard.

[Leiomyosarcoma and rhabdomyosarcoma: rare tumors of the kidney]. / Leiomiosarcoma y rabdomiosarcoma: tumores raros del riñón.

Renal sarcoma represents 2.8% of all renal malignant tumours. The most common kind is leiomyosarcoma while rhabdomyosarcoma is the most unusual. Clinical presentation and diagnosis of both types of tumours are unspecific. Treatment is controversial and the overall prognosis poor. The present paper presents two cases, one leiomyosarcoma and one rhabdomyosarcoma which were treated surgically and were followed by chemo- and radiotherapy. A revision of the criteria for grading these as primary tumours of the kidney is also included.

[Renal hematomas secondary to extracorporeal shockwave lithotripsy]. / Hematomas renales secundarios a litotricia extracorpórea por ondas de choque.

OBJECTIVE: 0.6% of the patients submitted to extracorporeal shock wave lithotripsy (ESWL) present symptomatic renal hematoma that can permanently alter renal function and morphology. The present study analyzes the risk factors, types of renal hematoma secondary to shock waves and treatment. METHODS: The study comprised 9 patients with ESWL-induced renal hematoma that had been diagnosed by renal ultrasound and/or urography. Patient evaluation included CT and an isotopic renogram. Two patients had associated arterial hypertension and ischemic coronary heart disease treated with antiaggregants. In 4 patients the renal calculi had produced partial obstruction of the urinary tract. RESULTS: In 4 patients the ESWL-induced renal hematoma was less than 1/3 of the renal volume and did not alter renal function. These patients have been classified as having type I renal hematoma and have been successfully managed conservatively. In 5 patients the hematoma was more than 1/3 of the renal volume and renal function was impaired. These patients have been classified as having type II renal hematoma. Spontaneous reabsorption of the hematoma facilitated recovery of renal function in one patient, two had persistent hematoma and progressive renal function impairment, and the remaining two patients underwent delayed evacuation of the hematoma on days 30-45. Decompression renal surgery preserved renal function of the two kidneys. CONCLUSION: Advanced arteriosclerosis, arterial hypertension, primary coagulation disorders or secondary to antiaggregation therapy and obstruction of the urinary tract increase the risk of ESWL-induced renal hematoma. Renal hematoma following ESWL has been classified as type I or type II. In the latter, renal decompression surgery facilitates recovery of renal function.

Inhibitory effects of N-acetylcysteine on superoxide anion generation in human polymorphonuclear leukocytes.

It has been suggested that reactive oxygen species released by activated polymorphonuclear leukocytes (PMN) in man is one mechanism of tissue injury. Therapeutic action aimed at increasing antioxidant defence mechanisms is still a clinical challenge. This study examines the activity of N-acetylcysteine, a known antioxidant, in the protection of PMN exposed in-vitro to the chemoattractant peptide fMet-Leu-Phe (FMLP), the protein kinase C activator phorbol myristate acetate or the lipid peroxidation promoter t-butyl hydroperoxide. FMLP (3-300 nM) and phorbol myristate acetate (160 pm-160 nM) induced concentration-related superoxide anion generation. Pre-treatment with N-acetylcysteine (33-333 microM) resulted in concentration-related inhibition of superoxide production induced by FMLP (30 nM) or phorbol myristate acetate (16 nM);-log IC50 values were 3.97 +/- 0.07 and 3.91 +/- 0.10, respectively. Changes in intracellular calcium ion concentration ([Ca2+]i) induced by FMLP (30 nM) were studied in fura-2-loaded human PMN. FMLP produced a transient calcium response, i.e. a peak followed by decay to a residual value above baseline. N-Acetylcysteine (333 microM) did not affect either basal [Ca2+]i values or changes in [Ca2+]i values after treatment with FMLP. Activation by phorbol myristate acetate caused a reduction in glutathione levels from 5.94 +/- 0.86 (control) to 1.84 +/- 0.51 nmol/3 x 10(6) cells (P < 0.05 compared with control). Pre-treatment with N-acetylcysteine (333 microM) fully reversed the reduction in glutathione levels induced by phorbol myristate acetate (4.83 +/- 0.68 nmol/3 x 10(6) cells; P > 0.05 compared with control). Exposure to t-butyl hydroperoxide (0.5 mM, 30 min) markedly increased malondialdehyde levels (from 0.03 +/- 0.02 to 0.73 +/- 0.07 nmol/10(6) cells), and index of lipid peroxidation. Malondialdehyde levels were significantly reduced in PMN treated with N-acetylcysteine (333 microM; 0.55 +/- 0.04 nmol/10(6) cells; P < 0.05 compared with untreated cells exposed to t-butyl hydroperoxide). In conclusion, N-acetylcysteine reduces superoxide generation in response to FMLP and phorbol myristate acetate and partially protects against lipid peroxidation in PMN from man. The protection afforded by N-acetylcysteine is not related to alteration of the intracellular calcium signal but might be effected by replenishment of the intracellular glutathione levels.

Leukotrienes and their antagonists in allergic disorders.

Cysteinyl leukotrienes are important mediators of asthma, and inhibition of their effects may represent a potential breakthrough in the therapy of allergic rhinitis and asthma. Strategies for inhibition of cysteinyl leukotriene receptors and inhibition of 5-lipoxygenase activity. The leukotrienes antagonists, with particular reference to asthma and allergic rhinitis, is reviewed in this paper. In studies in asthmatic patients, these compounds can inhibit bronchoconstriction in response to exercise, aspirin and allergen. Results from clinical studies using receptor antagonists, such as LY-171883, SK&F-104353, ICI-204219, ONO-1078, MK-751, MK-0679, demonstrate beneficial effects, with improvement in symptoms and forced expiratory volume in one second (FEV1), and a reduction in the use of beta 2-adrenergic relief medication. NZ-107 was studied for its effect on airway inflammation caused by intratracheal injection of LTB4 or IL-5, or by inhalation of PAF, and by cell activation. Analysis of the BAL fluid revealed that both induced eosinophilia and neutrophilia were suppressed. Surprisingly, although PAF and superoxide generation were inhibited in macrophages and eosinophils, NZ-107 had no effect on neutrophil activation. U-75302 was studied in guinea pigs and inhibited LTB4 induced chemotaxis of eosinophils in vitro and antigen-induced lung eosinophilia in vivo. Further studies are needed to clarify the exact mechanism by which these compounds provide beneficial effects.