[Supply medicinal products improvement in outpatient care in a hospital pharmacy service]. / Mejora de la dispensación a pacientes externos de un servicio de farmacia hospitalaria.

INTRODUCTION: Pharmaceutical care to outpatients is currently one of the main occupations of hospital pharmacy services (PEX). There are several questionnaires to measure the satisfaction of the PEX of a pharmacy service, and the results of these questionnaires can generate improvement actions that result in satisfaction. OBJECTIVES: To verify if a satisfaction questionnaire for outpatients is valid for the generation of improvements in the care provided, and if after its implementation, the same questionnaire is able to detect changes in satisfaction. MATERIAL AND METHOD: Prospective study of a single center carried out in a tertiary hospital in 2015 and 2016. A questionnaire previously validated with 16 Likert-type items was used. Demographic and classification data were collected. A descriptive analysis was performed and the internal consistency was calculated using the Cronbach's α value. RESULTS: A total of 258 questionnaires were collected in 2015 and 493 in 2016. There were no differences in the baseline characteristics of the patients and users of the service. The items with the lowest satisfaction scores in 2015 (comfort of the waiting room, dispensing privacy, drug pick-up time and medication pick-up time) guided the improvement actions to be implemented. In 2016 there was an improvement in the waiting time until collection in 12.3% (p = 0.002); in the comfort of the waiting room 4.9% (p = 0.304); business hours for medication collection, 10.7% (p = 0.013); and in the confidentiality of the dispensation 4% (p = 0.292). The remaining scores fluctuated minimally, with no statistical significance at all. A 5.1% improvement in overall satisfaction was found (p < 0.001). Satisfaction values obtained as a whole were high. CONCLUSIONS: The satisfaction questionnaire is a valid instrument for generating actions to improve the care received in an outpatient unit of a pharmacy service. This same questionnaire is a tool to monitor the changes implemented to improve the care received.

Aging affects but does not eliminate the enzymatic antioxidative response to hypoxia/reoxygenation in cerebral cortex.

The effect of aging on basal and hypoxia/reoxygenation levels of both oxidative stress (protein carbonyl and TBARS) and antioxidative-enzyme activity (Cu/Zn-SOD; Mn-SOD; Catalase, CAT; Se-independent and Se-dependent glutathione peroxidase, GPX; glutathione transferase, GST and glutathione reductase, GR) has been studied in the cerebral cortex of adult and old rats. Oxidative stress markers increased with aging and show an age-dependent post-hypoxic response. Moreover, aging caused either no change (GST, GR and CAT) or an increase (Se-GPX, Cu/Zn-SOD, Mn-SOD) in the basal activity of the enzymes analysed. Only Se-independent GPX activity decreases. However, we detected an age-dependent response of SODs to the hypoxic injury. The early and sustained Cu/Zn-SOD activity rise in adult animals became late and weak in aged animals. Meanwhile, aging slowed the Mn-SOD post-hypoxic response although this activity was consistently higher in aged rats. Aging eliminated the post-hypoxic CAT response, but, perhaps offset by increased GPX activity, did not affect the GST response and slightly reduced post-hypoxic GR activity. In conclusion, aging rise basal ROS production, does not diminish or even increase the antioxidative-enzyme activity, and may slow but does not usually eliminate the enzymatic antioxidant response to the increased post-hypoxic ROS generation.

Constitutive nitric oxide synthases are responsible for the nitric oxide production in the ischemic aged cerebral cortex.

Aged brain shows reduced biological plasticity to meet emergency conditions such as ischemia, a process in which nitric oxide (NO) and apoptosis have been shown to play important roles. Using a model of transient global ischemia, we have analyzed the NO system and the p53, bax and bcl-2 response in the cerebral cortex of aged rats. Although immediately after ischemia the NO level is maintained, the reperfusion period increases NO concentrations together with the following: (i) greater bulk-protein nitration mainly due to a 50-kDa immunoreactive band; (ii) an increase in p53 protein; and (iii) an up-regulation of Bax together with a down-regulation of Bcl-2. These results match up with induced endothelial nitric oxide synthase expression immediately after ischemia and in neuronal nitric oxide synthase with the reperfusion. However, inducible nitric oxide synthase was not altered with ischemia/reperfusion. Altogether, these data suggest that NO production in cerebral cortex of aged ischemic animals is due to the constitutive NO synthase isoforms. This response is accompanied by the increased expression of pro-apoptotic proteins.

Upregulation of endothelial nitric oxide synthase maintains nitric oxide production in the cerebellum of thioacetamide cirrhotic rats.

This study examines the expression and cellular distribution pattern of nitric oxide synthase (NOS) isoforms, nitrotyrosine-derived complexes, and the nitric oxide (NO) production in the cerebellum of rats with cirrhosis induced by thioacetamide (TAA). The results showed local changes in the tissue distribution pattern of the NOS isoforms and nitrated proteins in the cerebellum of these animals. Particularly, eNOS immunoreactivity in perivascular glial cells of the white matter was detected only in TAA-treated animals. In addition, although neither neuronal NOS (nNOS) nor inducible NOS (iNOS) cerebellar protein levels appeared to be affected, the endothelial NOS (eNOS) isoform significantly increased its expression, and NO production slightly augmented in TAA-treated rats. These NOS/NO changes may contribute differently to the evolution of the hepatic disease either by maintaining the guanosine monophosphate-NO signal transduction pathways and the physiological cerebellar functions or by inducing oxidative stress and cell damage. This model gives rise to the hypothesis that the upregulation of the eNOS maintains the physiological production of NO, while the iNOS is silenced and the nNOS remains unchanged. The differential NOS-distribution and expression pattern may be one of the mechanisms involved to balance cerebellar NO production in order to minimize TAA toxic injury. These data help elucidate the role of the NOS/NO system in the development and progress of hepatic encephalopathy associated with TAA cirrhosis.

Immunohistochemical localisation of neuronal nitric oxide synthase in the rainbow trout kidney.

The distribution of nitrergic nervous structures in the trout kidney was studied by peroxidase-linked ABC immunostaining procedures using a polyclonal antibody raised against the neuronal isoform of nitric oxide synthase. The nitrergic plexus reaches the kidney along the vasculature, mainly running with the postcardinal vein where nitrergic fibres, microganglia like cellular clusters and isolated neurones were detected. The atubular head-kidney only showed isolated nitrergic fibres close to the larger arteries. On the other hand, the collecting tubules, collecting ducts, large arteries and glomerular arterioles of the tubular middle and posterior trunks were innervated by nitrergic fibres even though immunoreactive neurones were also observed in close apposition to some tubular elements and large arteries. These results suggest that, according to morphofunctional differences between the fish and mammalian kidneys, nitrergic neural structures may be involved in the control of particular renal functions in the rainbow trout.

Quantitative and ultrastructural changes in glia and pericytes in the parietal cortex of the aging rat.

The frequency of astrocytes, microglia plus oligodendrocytes, and pericytes displaying nuclei was analyzed and quantified in 160-microm-wide strips of the parietal cortex (Par1 region) from young and aged Wistar rats. The study was performed on two groups of rats aged 3-4 and 32-36 months. Quantifications of the glial cell types and pericytes were made in 1-microm-thick sections stained with toluidine blue. Ultrathin sections were also made to analyze the ultrastructural features of these cells during aging. Astrocytes and pericytes increased in number by about 20% and 22%, respectively, with age. These increases were most significant in layers II-IV and V for both cellular types. Clusters of astrocytes were common in these layers of aging rats. The ultrastructural analysis also indicated changes in all cell types that stored inclusions and vacuoles with age, which were particularly abundant in microglial cells. End-feet astrocytes and pericytes surrounding the vascular wall also contained vacuoles and inclusions, and consequently the vascular wall increased in thickness. In conclusion, the aging process increased astrocyte and pericyte populations, but not microglia plus oligodendrocyte populations, in the rat parietal cortex. Although no significant change in nuclear size could be observed in any cell type, all glial cells as well as pericytes underwent morphological ultrastructural changes. These modifications may result from the need to correct possible homeostatic imbalances during aging.

Neuronal and inducible nitric oxide synthase and nitrotyrosine immunoreactivities in the cerebral cortex of the aging rat.

Neuronal and inducible nitric oxide synthase (nNOS and iNOS) and nitrotyrosine immunoreactivities were localized and semiquantitatively assessed in the cerebral cortex of aged rats by means of light microscopic immunocytochemistry and Western blotting, using a new series of specific polyclonal antibodies. In the aged rats the strongly nNOS-immunoreactive multipolar neurons found in layers II-VI of the cortex of young rats were seen in similar numbers, but showed varicose, vacuolated, and fragmented processes, with an irregular outline and loss of spines. A large number of more weakly nNOS-positive neurons, characterized by a ring of immunoreactive cytoplasm, and not seen in young rats, were observed in layers II-VI of aged rat cortex. While no iNOS-immunopositive neurons were found in the cortex of young rats, a large number of such neurons appeared throughout the aged rat cortex. Nitrotyrosine-positive cells outnumbered total NOS-positive neurons in the cortex of young rats, but this relation was inverted in the aged rats, although these showed a slight increase in the number and staining intensity of nitrotyrosine-positive cells. Western blots of brain extracts showed a several-fold increase in both nNOS- and iNOS-immunoreactive bands in the aged rat, but a less marked increase in nitrotyrosine-containing proteins. The results suggest that while nNOS and iNOS expression is substantially increased in the aged rat cortex, this is not necessarily accompanied by a proportionate increase in nitric oxide synthesis. The mechanisms underlying the increased expression of nNOS and iNOS, and the functional implications of this increase, require elucidation.

Nitrergic innervation of the cat liver.

The aim of this work was to study the nitrergic innervation in the liver of the cat using immunocytochemical procedures. At the hepatic hilus, a rich plexus of neuronal nitric oxide synthase immunoreactive (nNOS-IR) nerve fibers and ganglia was detected around the interlobular branch of the bile duct. nNOS-IR nerve fibers were observed running with the components of the intralobular portal triads located close to the hepatic hilus, as well as with a few vessels and ducts of the deeper parenchyma. These latter fibers, beside others located in Glisson's capsule, occasionally showed short ramifications entering the parenchyma itself. The present results suggest that, in the cat liver, nNOS is involved in the autonomic control of hepatic blood flow, with a limited role in the regulation of hepatobiliary excretory activity and hepatocellular metabolic function.

Distribution of neuronal nitric oxide synthase in the rat liver.

We studied the distribution of neuronal nitric oxide synthase (nNOS) in the rat liver with a specific polyclonal antibody by using immunocytochemical procedures in the light microscopic level. Immunoreactive varicose nerve fibers were found forming a dense plexus around the interlobular hepatic artery and the interlobular bile duct in the hepatic hilus, and in the hepatic artery ramifications of the portal triads. The density of nNOS positive nerve fibers decreases with successive portal ramifications, and some non-immune positive nerve fibers were found in the distal portions of the arterial vessels. The presence of the nNOS positive nerve fibers suggests that the possible main functional role could be related with the regulation of hepatic blood circulation and hepatobiliary activities.

Effect of thioacetamide and dexamethasone on serum lipids in rats fed on high-fat sunflower or olive oil diets.

We have previously reported that high-fat diets develop hepatic steatosis and, depending on the fat quality, affect serum lipid levels differently (J Nutr Sci Vitaminol, 1997, 43, 155-160). The aim of this work is to study the influence of high-fat diets (14% sunflower or olive oils) on serum lipids in a model of hepatic acute damage induced by thioacetamide, and their influence when dexamethasone is administered before thioacetamide injection. Serum lipids and hepatic collagen have been evaluated using biochemical methods, and the steatotic process by histological staining. The results showed that hepatic steatosis and fibrosis are developed either by high-fat diets or thioacetamide injection. Pretreatment with dexamethasone did not decrease the hepatic collagen content. Thioacetamide injection alone or pretreatment with dexamethasone produced increase in serum tryglicerides (TG), total cholesterol (TC) and LDL-C in both high-fat diet groups, and a HDL-C increase in the olive-oil group, even though the atherogenic indices (HDL/TC and HDL/TG) were different depending on the enriched diet. The administration of high-fat diets to study the influence of the fat quality on health and disease should be interpreted carefully due to the ability of the diets themselves to cause hepatic damage.

High-fat sunflower and olive oil diets affect serum lipid levels in steatotic rat liver differently.

This work describes the long-term effects of two different diets, one rich in olive oil and the other in sunflower oil, on serum lipid and lipoprotein levels after the establishment of fatty liver in rats 8 and 15 months old. The serum lipid and lipoprotein levels as well as the steatotic process have been evaluated by biochemical and histological methods, respectively. The results showed that fatty liver was well developed with both long-term high-fat diets, and hepatocytes were filled with many lipid droplets. This process was more evident in the portal zones, where fat hepatocytes were more numerous. Serum total cholesterol (TC) and HDL-C levels were highest in the sunflower oil fed rats, whereas the TG and LDL-C levels were highest in the olive oil group. Finally, the atherogenic indexes (HDL/TC, HDL/LDL, HDL/(TC-HDL)) were higher in the sunflower oil diet group than in the olive oil group.

[Aging and neurodegeneration: molecular and cellular bases]. / Envejecimiento y neurodegeneración: bases moleculares y celulares.

OBJECTIVE: A review about the possible cellular and molecular mechanisms of aging and related neurodegenerative diseases. DEVELOPMENT: The mechanisms involved in neuronal decrease, connectivity losses and glial reactivity, detected both in neurodegenerative (Alzheimer's disease) and physiological aging, are analyzed from the morphological and histological point of view to provide the morphofunctional base of the cognitive and intellectual alterations characterizing the senescence process. Taken together, these data are correlated to the possible genetical aspects implied in this process, reviewing the most relevant results on senescence and cellular death obtained from yeast, fruit fly and nematodes; besides this, a brief review of the molecular biology of gerontogenes was carried out, and the possible mechanisms inducing aging and neurodegenerative processes are analyzed according to the state-of-the-art related theories. Finally, cellular, biochemical and genetical data are correlated in the signal transduction way implied in the increase of the intracellular calcium level as the starting point of cell death. CONCLUSIONS: The main process implied in the neuronal cell death responsible for aging and the related neurodegenerative diseases are started by different agents such as the lacking of neurotrophic factors, hypoxia, hypoglycemia, excitotoxicity, and oxygen and nitrogen free radicals.

Distal urethrectomy for localized penile squamous carcinoma in situ extending into the urethra: an updated series.

INTRODUCTION: The presence of squamous carcinoma in situ (CIS) of the distal penis extending into the urethral meatus is generally considered a contraindication for glans-sparing procedures. Distal urethrectomy with subsequent reconstruction can provide an alternative approach toward urethral resection while providing penile preservation in select cases. Unfortunately, long-term oncologic outcomes with this approach are ill-defined. MATERIALS AND METHODS: Between 1988 and 2012, five patients at Indiana University Medical Center underwent distal urethrectomy with reconstruction for penile squamous CIS extending into the urethral meatus. This cohort was retrospectively reviewed to evaluate functional and oncological outcomes. RESULTS: Of the five patients, four presented with glanular lesions and were initially managed with Mohs procedure in three cases, and local excision in one. The final patient presented with extensive urethral disease and was managed with primary urethrectomy. Reconstruction was performed with penile skin pedicle grafts in four patients and perineal urethrostomy in one. Final pathologic stage was T1 in one patient and Tis in the remaining four. Follow-up ranged from 6 to 96 months. One local recurrence was verified; however, it occurred outside the urethral area. This was confirmed in the pathologic analysis after the patient underwent a partial penectomy. Meatal dilation was necessary in two patients 12 and 7 months after the procedure. CONCLUSION: Distal urethrectomy for penile squamous CIS extending into the urethral meatus is a valid alternative to achieve negative surgical margins while preserving a penile function. Oncologic outcomes appear acceptable but larger series are still warranted to confirm our findings.

Age and sex-related serum changes in nitric oxide: correlations with serological markers.

Serum nitric oxide levels, systematically determined in 200 men and women from 18 to 65 year-old, undergo age and sex changes that strongly correlate with serological markers such as those related with cardiovascular functions and lipid profile.

Quantitative morphological changes in neurons and glia in the frontal lobe of the aging rat.

The quantitative morphological changes in neurons and glia during the aging process were analyzed in the different cortical layers, grouped as I, II-IV, V, and VI, of the frontal cortex of the rat. The parameters analyzed were cortical volume, neuronal density, glial density, and neuronal soma and nucleus areas. No changes with age were found in the volume of the layers, in neuronal density (with the exception of layer I), or in the area of the neuronal soma. However, older animals showed a 10 to 20% increase in glial density, depending on the layer studied. In addition, there was an age-related decrease in the area of the neuronal nucleus in layers II-IV, V, and VI. These results support the idea that the aging frontal cortex undergoes structural changes that may be involved in the morphological basis of memory and cognitive impairments characteristic of aging.

Vegetative innervation of the esophagus. III. Intraepithelial endings.

Intraepithelial fibers do occur in the mucosa of the esophagus, as demonstrated by the osmium tetroxide-zinc iodide method in cats and rhesus monkeys. The esophagus is divided into three parts, in order to study the penetration incidence, and the uppermost and the lowest show the greatest density of penetration, while in the middle portion only occasional fibers in small numbers are found. The specific characteristics observed in this type of fiber, such as their distribution along the wall of the esophagus, the levels reached by their endings within the mucous epithelium itself and this same epithelium considered as the specified destination of the endings, lead to the belief that they may be functionally regarded as structures of a sensory character.

Vegetative innervation of the esophagus. II. Intraganglionic laminar endings.

The intraganglionic laminar endings in the esophagus of the cat and the rhesus monkey show absolute equivalence between the results in both species from the morphological standpoint. The different types of apparatus found are described, with their location in the esophagus and their percentage distribution in relation to the different portions of its wall. The osmium tetroxide-zinc iodide technique gives pictures equivalent to those using silver impregnations, with the added advantage that the former brings out the morphological details more clearly, to the point of showing up the peculiar characteristics of the edges with their thorn-like protrusions. The complete independence of these structures within the ganglion is confirmed, and evidence is provided for rejecting the possibility that they might be dendritic prolongations of the neuronal elements composing the intramural ganglia. A possible afferent function is proposed, which, however, must be considered an open question, pending the results of further experimental investigation.

Vegetative innervation of the esophagus. IV. Endings in the tela submucosa and tunica muscularis.

The osmium tetroxide-zinc iodide technique has been applied to the study of the tela submucosa and tunica muscularis in the esophagus of the cat and the rhesus monkey. Other than the classical terminal vegetative formation, three types of endings were found, classified as perifascicular, free and intramuscular. The comparative results between cat and rhesus monkey show differences in the distribution and number of such endings. A sensory functional significance is proposed and discussed.

Sensorivagal nature of oesophageal submucous layer nerve endings. Determination of surgical degeneration methods.

Two types of surgery were performed on domestic cats to establish the source and functional significance of the apparatuses we have denominated perivascular, perifascicular and free endings of the oesophageal submucous layer. After extirpation of the cranial regions of the sympathetic ganglionic chain, of all the cervical ganglia and of the four proximal thoracic ganglia, no Wallerian degeneration of the concerned nerve endings was observed, thus ruling out their dependence on this neural pathway. After ablation of the central portion of the nodose ganglion, of the vagal nerve. Wallerian degeneration was observed in these nerve-ending apparatuses, thus demonstrating: (i) that these nerve apparatuses depend on perikarya situated in the nodose ganglion and (ii) that they are sensory receptors.

[Ganglionic cytoarchitecture of the esophagus in the cat and the rhesus monkey]. / Quelques aspects de la cytoarchitectonie ganglionnaire de l'oesophage chez le chat et le singe rhésus

The comparative study of neuron distribution in the myenteric and submucous plexuses has granted an appraisal of the various neuron types in the oesophagus ganglia of cat and rhesus monkey. Neurons are classified in Dogiel types I and II, plus an intermediate one, the so-called "type III". From the functional point of view, the hypothesis is established that these types are, respectively: efferent, afferent and associative. A confirmation of this hypothesis requires the endorsement of the results of degeneration experiments and their integration with the roles of the rest of the structures which participate in the oesophagus innervation as a whole.