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Biomedical subjects

L Medina

Publications and source records attributed to L Medina.

At least 37 records · Page 2Linked to original sources

Immunohistochemical localization of DARPP32 in striatal projection neurons and striatal interneurons in pigeons.

DARPP32 is a D1-receptor associated signaling protein found in striatal projection neurons in mammals, including both substance P-containing (SP+) neurons and enkephalinergic (ENK+) projection neurons. The present study used immunohistochemical single- and double-labeling to examine the cellular localization of DARPP32 in pigeon striatum. Single-label studies revealed that DARPP32 is present in numerous medium-sized striatal perikarya and DARPP32+ axons and terminals were seen to profusely innervate the two major striatal projection targets, the pallidum and the substantia nigra. The single-labeling studies indicated that about 60% of all striatal perikarya labeled for DARPP32+ in striatum, which exceeds the abundance of either SP+ or ENK+ perikarya. Single-labeling studies also showed that the abundance of DARPP32+ fibers and terminals in pallidum exceeds that of either SP+ or ENK+ fibers and terminals in pallidum. Double-labeling found that 30-50% of striatal SP+ perikarya and 7-24% of ENK+ striatal perikarya labeled for DARPP32 in pigeon, and confirmed that DARPP32 was found in both SP+ and ENK+ fibers and terminals in pallidum. In contrast to its prevalence in striatal projection neurons, DARPP32 was virtually absent from cholinergic and NPY+ striatal interneurons, as also true in mammals. Our data are consistent with the interpretation that many SP+ neurons and many ENK+ neurons in avian striatum possess D1-type dopamine receptors and use a DARPP32 signalling pathway, although this may be more common for SP+ than for ENK+ neurons.

Animals↗

Calf thymus high mobility group proteins are nonenzymatically glycated but not significantly glycosylated.

Over the past decade, there have been many reports suggesting the presence of complex carbohydrates on nuclear and cytoplasmic proteins in mammalian cells. Some of the most often cited of these reports deal with the glycosylation of the high mobility group (HMG) proteins. These are relatively abundant chromosomal proteins that are known to be associated with nucleosomes and actively transcribed regions of chromatin. The original report describing HMG protein glycosylation presented several lines of evidence suggesting that these proteins are glycosylated, including carbohydrate compositional analysis and periodic-acid Schiff staining. We have attempted to repeat these observations with more highly purified protein than was utilized in the original study. Using carbohydrate compositional analysis performed by high pH anion exchange chromatography coupled to pulsed-amperometric detection, we saw no evidence for significant glycosylation of these proteins. In addition, we found no evidence for the presence of O-GlcNAc, a well known form of nuclear glycosylation. The HMG proteins did react with periodate, suggesting the presence of a modification containing cis-diols on the protein. Several tryptic peptides isolated from HMG 14 and 17 which retained the periodate reactivity had in common lysine residues, suggesting a potential modification of the straightepsilon-amino groups of lysines such as nonenzymatic glycation. Western blot analysis of the HMG proteins using anti-advanced glycation endproducts (AGE) antibodies confirmed the presence of glycation products on the HMG proteins.

Acetylglucosamine↗

SV40 large T antigen is modified with O-linked N-acetylglucosamine but not with other forms of glycosylation.

SV40 large T antigen has been reported to be modified with several different sugars including N-acetylglucosamine, galactose, and mannose. In this report we have reexamined the glycosylation of T antigen and found that while we could detect modification with N-acetylglucosamine, we could not detect any other sugars on the protein. Surprisingly, even though [3H]galactose could be metabolically incorporated into the protein, analysis showed that all of the radioactivity in T antigen had been converted to other species. The N-acetylglucosamine was demonstrated to be linked to the protein in the form of O-linked N-acetylglucosamine, the best characterized form of nuclear and cytoplasmic glycosylation in mammalian systems. We have localized the major site of glycosylation to the amino terminal portion of the molecule. Analysis of mutated T antigen where serines 111/112 were substituted with alanine suggest that these residues constitute a glycosylation site on the protein. These two serines fall within a typical O-linked N-acetylglucosamine glycosylation site (PSS) and are also known to be phosphorylated. Thus, it is likely that competition between phosphorylation and glycosylation occurs at this site.

Acetylglucosamine↗

Absorption studies of albendazole and some physicochemical properties of the drug and its metabolite albendazole sulphoxide.

In several studies of patients with neurocysticercosis under treatment with albendazole the pharmacokinetic data were difficult to interpret, probably because of slow and erratic drug dissolution response and absorption problems in-vivo. Because there is no information available about the physicochemical properties of the drug, the aim of this work was to explain this erratic behaviour by fully characterizing the solution behaviour of the drug and its metabolite. To accomplish this, the physicochemical properties, pKa and solubility, and in-vitro plasma binding of albendazole and its main metabolite, albendazole sulphoxide, were studied by conventional methods. The intestinal and gastric absorption and dissolution behaviour of albendazole were also studied. The solubility of both compounds is very low. Both are amphoteric molecules with two ionization steps, with pKa values of 10.26 and 2.80 for albendazole and 9.79 and 0.20 for albendazole sulphoxide; low pKa values were obtained by performing linear free energy relationship calculations. On the other hand, protein binding studies showed that albendazole is 89-92% bound to plasma proteins whereas for albendazole sulphoxide the figure is 62-67%. This metabolite is bound by albumin and to alpha1-glycoprotein. Absorption of albendazole occurs along the gastrointestinal tract and is limited by its solubility. Good dissolution profiles were observed when 0.1 M HCl was used as dissolution medium. The results show that 0.1 M HCl enables discrimination between the drug-release characteristics of different products.

Albendazole↗

Pharmacokinetic interaction in rabbits between a new anticonvulsant, DL-3-hydroxy-3-ethyl-3-phenylpropionamide, and phenytoin.

The effect of phenytoin on the disposition of DL-3-hydroxy-3-ethyl-3-phenylpropionamide (HEPP) has been studied in New Zealand white rabbits. Plasma HEPP levels decreased when the drug was administered with phenytoin. The area under the plasma concentration-time curve was reduced by 56.49% (from 43.23+/-7.0 to 18.81+/-2.03 microg h mL(-1)), the elimination half-life was also significantly (P<0.01) reduced (from 2.68+/-0.35 to 1.04+/-0.07 h) and the clearance was increased (from 0.35 to 0.81 L h(-1) kg(-1)). In-vitro protein binding to bovine serum albumin (BSA) and plasma was evaluated by equilibrium dialysis. Plasma protein binding was low (between 33.69 and 37.43% at concentrations ranging from 6.25 to 100 microg mL(-1)). The compound binds preferentially to albumin with an association constant (Ka) of 3.81 x 10(3) M(-1) at 37 degrees C. The results suggest a pharmacokinetic interaction between phenytoin and HEPP, probably on the drug-metabolizing enzyme system in the liver.

Animals↗

[Perforations of nasal septum: surgical considerations].

Perforations of the nasal septum vex both patients and physicians. A variety of methods have been devised to close septal perforations although none has gained universal acceptance. The following report presents a new approach for closing septal perforations using a modification of the Fairbanks technique. The "open" approach is performed as it provides easier access. A supporting graft is only used in the temporalis fascia, not in the cranial periosteum. Furthermore, the graft is sutured only on one side of the septum. Forty patients were treated with this technique and good results were obtained with small and mid-sized perforations (respectively 80% and 83%). In cases of large perforations the results were good enough (37.5%) to warrant recommendation of this technique.

Adult↗

[Clinical features and diagnosis of mucocutaneous leishmaniasis in patients of an endemic area in Salta].

A total of 39 patients with a clinical diagnosis of mucocutaneous leishmaniasis, in an endemic area for leishmaniasis in Salta, Argentina, were examined between June 1990 and December 1992. Of these cases, 87% (34/39) presented the cutaneous simple form, 10.3% the cutaneous multiple form and 2.6% the mucosal form. Lesions were more frequently located in legs and arms (71.8%), followed by trunk and multiple location (10.3%). Of the patients, 43% were housewives, students or children, suggesting that the infection could be contracted in the domestic or peridomestic environment. Of 39 patients diagnosed, in 22 (56.4%) the parasite was found. Direct microscopy (smear) permitted a diagnosis in 13 (59.4%) of these 22 patients. Among these, 5 (22.7%) had positive diagnosis by culture, and 9 (40.9%) by inoculation in hamsters. Ten parasite isolates (45.4%) were obtained. The smear is recommended as a diagnostic method for epidemiological surveillance due to the sensibility demonstrated herein and its easy application in the endemic area. The time of clinical evolution, from the appearance of the lesion up to the detection of the patient by Sanitary Agents, was approximately 90 days. This would be related to the frequency of the visits, usually every 3 months. Only one of 30 treated patients had a relapse at 6 months, due to non fulfillment of the treatment.

Adolescent↗

Evidence for a possible avian dorsal thalamic region comparable to the mammalian ventral anterior, ventral lateral, and oral ventroposterolateral nuclei.

In the present study, we investigated whether a dorsal thalamic region comparable to the motor part of the mammalian ventral tier (the ventral anterior nucleus, the ventral lateral nucleus, and the oral ventroposterolateral nucleus) exists in pigeon. With this aim, we reinvestigated the projections of the pigeon dorsal pallidum to the dorsal thalamus by using 1) injections of the anterogradely transported form of biotinylated dextran amine (BDA; 10,000 molecular weight) in the pigeon dorsal pallidum (paleostriatum primitivum) and 2) injections of the retrogradely transported form of BDA (3,000 molecular weight) in the pigeon dorsal thalamus. Our results indicate that the dorsal pallidum in pigeons projects to three areas of the dorsal thalamus: the dorsointermediate posterior nucleus, the ventrointermediate area, and the nucleus subrotundus. Only the projection to the dorsointermediate posterior nucleus was described previously (Karten and Dubbeldam [1973] J. Comp. Neurol. 148:61-90; Kitt and Brauth [1982] Neuroscience 6:1551-1566). To investigate whether any of the dorsal thalamic nuclei receiving pallidal input project to a motor cortical field, injections of the retrograde tracer Fluoro-Gold were placed into the rostral Wulst. This is an avian cortical field that appears to contain a region comparable to mammalian primary somatomotor cortex (Karten [1971] Anat. Rec. 169:353; Wild [1992] J. Comp. Neurol. 287:1-18). Our results indicate that neurons in the rostral ventrointermediate area, but not in the nucleus subrotundus, the dorsointermediate posterior nucleus, or the intermediate or caudal parts of the ventrointermediate area, project to the rostral Wulst. In addition to the input from the dorsal pallidum, the avian ventrointermediate area also receives input from the lateral substantia nigra and the lateral and internal cerebellar nuclei (present results). Our results suggest the existence in birds of a pallidothalamocortical loop similar to the pallidoventral tier-motor cortex loop of mammals and suggest that the avian ventrointermediate area is comparable to the motor part of the mammalian ventral tier in both location and connections. If this is confirmed by physiological experiments, then it would indicate that basal ganglia control of movement mediated by a pallidothalamocortical loop may have evolved with the stem reptiles.

Animals↗

The efferent projections of the dorsal and ventral pallidal parts of the pigeon basal ganglia, studied with biotinylated dextran amine.

In the present study we have investigated the efferent projections of both the dorsal and the ventral pallidum of the pigeon basal ganglia, using the sensitive anterograde tracer biotinylated dextran amine [Veenman C. L. et al. (1992) J. Neurosci. Meth. 41, 239-254]. Injections of biotinylated dextran amine in the pigeon dorsal pallidum produced numerous fibers and terminals in specific nuclei of the thalamus, hypothalamus, pretectum and midbrain tegmentum. In the thalamus, labeled fibers and terminals were observed in the avian thalamic reticular nucleus, the proposed motor part of the avian ventral tier (ventrointermediate area), the avian parafascicular nucleus (nucleus dorsointermedius posterior), as well as in the avian nucleus subrotundus (which may be comparable to the posterior intralaminar nuclei of mammals). Labeled fibers and terminals were also observed in the avian subthalamic nucleus (anterior nucleus of the ansa lenticularis), in the pretectum (nucleus spiriformis lateralis) and in the avian substantia nigra pars reticulata. Injections of biotinylated dextran amine in the pigeon ventral pallidum produced fibers and terminals in specific centers of the telencephalon, hypothalamus, thalamus, epithalamus, and midbrain and isthmic tegmentum. Labeled fibers and terminals were also observed in the avian subthalamic nucleus and the inmediately adjacent lateral hypothalamus, the avian thalamic reticular nucleus, the avian medidorsal nucleusaand posterior intralaminar nuclei, and the lateral habenula. Finally, labeled fibers and terminals were found in the ventral tegmental area, the avian substantia nigra pars compacta and the midbrain/isthmic tegmentum, which includes the pedunculopontine tegmental nucleus. Our results indicate that both the dorsal and ventral pallida of birds have unique and specific projection patterns, which are very similar to those of their counterparts in mammals. Our study suggests that these avian basal ganglia regions may be related mainly to somatomotor and limbic functions, respectively.

Anatomy, Artistic↗

Avian homologues of mammalian intralaminar, mediodorsal and midline thalamic nuclei: immunohistochemical and hodological evidence.

This paper presents and reviews data suggesting that the dorsal thalamic zone (abbreviated DTZ) in birds is homologous to the intralaminar, midline, and mediodorsal thalamic nuclear complex (abbreviated IMMC) in mammals. The DTZ is located dorsomedially in the diencephalon of birds and consists of several subnuclei: nucleus dorsomedialis anterior thalami (DMA), nucleus dorsomedialis posterior thalami (DMP), nucleus dorsolateralis anterior thalami, pars medialis (DLM), nucleus dorsointermedius posterior thalami (DIP), nucleus dorsolateralis posterior thalami (DLP), and nucleus subhabenularis lateralis (SHL). Our immunohistochemical studies show that: (1) SHL and medial and dorsal parts of DMA and DMP are relatively rich in GABAergic, enkephalin-containing, substance P-containing, and cholinergic fibers; (2) lateral parts of DMA and DMP are relatively poor in these neurotransmitters; and (3) DIP, DLP, and DLM are moderately rich in cholinergic and substance P-containing fibers. Our retrograde pathway tracing studies indicate that the DIP and DLP in the more lateral parts of DTZ project to somatic striatum, while the DMA, DMP, and SHL located more medially in the DTZ project to visceral/limbic striatum. Our anterograde tracing studies indicate that DIP receives afferents from the dorsal pallidum, whereas DMA and DMP appear to receive afferents from both the ventral striatum and ventral pallidum. Diverse prior studies have shown that in general medial and lateral components of DTZ are connected with visceral/ limbic and somatic brain regions, respectively. These characteristics indicate that: (1) SHL and medial and dorsal parts of DMA and DMP are comparable to mammalian midline thalamic nuclei, including the medial components of the intralaminar nuclei; (2) lateral parts of DMA and DMP are comparable to the mediodorsal nucleus in mammals; (3) DIP is comparable to the parafascicular nucleus in mammals; and (4) DLM and DLP are comparable to the laterally located intralaminar nuclei in mammals. The comparability of avian DTZ and mammalian IMMC suggests that they evolved from thalamic precursor nuclei present in the common reptilian ancestors and that they may perform similar roles in the movement control function of the basal ganglia.

Animals↗

Light and electron microscopic immunohistochemical study of dopaminergic terminals in the striatal portion of the pigeon basal ganglia using antisera against tyrosine hydroxylase and dopamine.

A dopaminergic projection from the midbrain to the striatal portion of the basal ganglia is present in reptiles, birds, and mammals. Although the ultrastructure of these fibers and terminals within the striatum has been studied extensively in mammals, little information is available on the ultrastructure of this projection in nonmammals. In the present study, we used immunohistochemical labeling with antibodies against tyrosine hydroxylase (TH) or dopamine (DA) to study the dopaminergic input to the striatal portion of the basal ganglia in pigeons (i.e., lobus parolfactorius and paleostriatum augmentatum). At the light microscopic level, the anti-TH and anti-DA revealed a similar abundance and distribution of numerous labeled fine fibers and varicosities within the striatum. In contrast, the use of an antidopamine beta-hydroxylase antiserum (which labels only adrenergic and noradrenergic terminals) labeled very few striatal fibers, which were restricted to visceral striatum. These results demonstrate that anti-TH mainly labels dopaminergic terminals in the striatum. At the electron microscopic level, the anti-TH and anti-DA antisera labeled numerous axon terminals within the striatum (15-20% of all striatal terminals). These terminals tended to be small (with an average length of 0.6 microns) and flattened, and their vesicles tended to be small (35-60 nm in diameter) and pleomorphic. About 50% of the terminals were observed to make synaptic contacts in the planes of section examined, and nearly all of these synaptic contacts were symmetric. Both TH+ and DA+ terminals typically contacted dendritic shafts or the necks of dendritic spines, but a few contacted perikarya. No clear differences were observed between TH+ and DA+ terminals within medial striatum (whose neurons project to the nigra in birds) or between TH+ and DA+ terminals within lateral striatum (whose neurons project to the pallidum in birds). In addition, no differences were observed between medial and lateral striata in either TH+ or DA+ terminals. Thus, there is no evident difference in pigeons between striatonigral and striatopallidal neurons in their dopaminergic innervation. Our results also indicate that the abundance, ultrastructural characteristics, and postsynaptic targets of the midbrain dopaminergic input to the pigeon striatum are highly similar to those in mammals. This anatomical similarity is consistent with the pharmacologically demonstrable similarity in the role of the dopaminergic input to the striatum in birds and mammals.

Animals↗

Calretinin is largely localized to a unique population of striatal interneurons in rats.

Previous studies have reported the presence of the calcium binding protein calretinin in neurons in the striatal part of the basal ganglia in rats and primates. In the present study, immunofluorescence double-labeling techniques and immunofluorescence combined with retrograde labeling were used in rats to determine whether calretinin is found in any of the known types of striatal neurons. The results showed that a small fraction of the calretinin-containing neurons (< 10%) contain parvalbumin, but none of the calretinin-containing striatal neurons contained markers for the other two major types of striatal interneurons (i.e., choline acetyltransferase-containing cholinergic neurons and somatostatin-containing neurons). Additionally, calretinin was not found in projection neurons, using either calbindin or DARPP32 as immunofluorescent markers of striatal projections neurons in general, or using retrograde labeling to specifically identify either striatonigral or striatopallidal neurons. Thus, calretinin appears to be largely found in a unique population of striatal interneurons in rats. This population appears to be about one third the abundance of any of the previously identified populations of striatal interneurons.

Animals↗

Differential abundance of superoxide dismutase in interneurons versus projection neurons and in matrix versus striosome neurons in monkey striatum.

To investigate whether differences in vulnerability to free radicals might underlie differences among striatal neurons in their vulnerability to neurodegenerative processes such as occur in ischemia and Huntington's disease, we have analyzed the localization of superoxide free radical scavengers in different striatal neuron types in normal rhesus monkey. Single- and double-label immunohistochemical experiments were carried out using antibodies against the enzymes copper, zinc superoxide dismutase (SOD1), or manganese superoxide dismutase (SOD2), and against markers of various striatal cell types. Our results indicate that the striatal cholinergic and parvalbumin interneurons are enriched in SOD1 and/or SOD2, whereas striatal projection neurons and neuropeptide Y/somatostatin (NPY+/SS+) interneurons express only low levels of both SOD1 and SOD2. We also found that projection neurons of the matrix compartment express significantly higher levels of SOD than those in the striosome compartment. Since projection neurons have been reported to be more vulnerable than interneurons and striosome neurons more vulnerable than matrix neurons to neurodegenerative processes, our results are consistent with the notion that superoxide free radicals are at least partly involved in producing the differential neuron loss observed in the striatum following global brain ischemia or in Huntington's disease.

Animals↗

Differential abundance of glutamate transporter subtypes in amyotrophic lateral sclerosis (ALS)-vulnerable versus ALS-resistant brain stem motor cell groups.

Previous studies have suggested that defective high-affinity glutamate uptake, due mainly to a major loss of the astroglial-specific GLT-1 glutamate transporter, underlies the selective motoneuron degeneration observed in sporadic ALS (24, 28). If a defect in glutamate transport underlies the pathogenesis of sporadic ALS, the glutamate transporter subtype found to be lost in sporadic ALS should be present in abundance in the affected motor nuclei under normal conditions. To investigate this, we used immunohistochemical methods to analyze the localization of two subtypes of high-affinity glutamate transporters in the cranial motor nuclei of normal monkey brain stem: GLT-1, localized to astroglia; and EAAC1, localized to neurons. Our results indicated that all motor cell groups of monkey brain stem are rich in the GLT-1 glutamate transporter, which is localized to astroglial cells and processes that surround and envelop motoneuron cell bodies and dendrites. Image analysis indicated that the abundance of GLT-1 immunoreactive astroglial elements in ALS-vulnerable motor cell groups (i.e., the trigeminal, facial, and hypoglossal motor cell groups) is higher than in ALS-resistant motor cell groups (i.e., the oculomotor, trochlear, and abducens motor cell groups), and statistical analysis showed that this difference is significant. Our results also indicated that both ALS-vulnerable and ALS-resistant motor cell groups of monkey brain stem are relatively poor in EAAC1 immunoreactivity. Therefore, in the case of a loss in the GLT-1 glutamate transporter in sporadic ALS, glutamate may increase in the vicinity of motoneurons in all brain-stem motor cell groups, but especially in the ALS-vulnerable motor cell groups, which are normally richer in GLT-1. Increased extracellular glutamate could lead to excess entry of Ca2+ into motoneurons via glutamate-gated or voltage-activated Ca2+ channels and produce degeneration of those motoneurons unable to resist the insult. Since motoneurons in the ALS-resistant motor cell groups of the brain stem are enriched in the Ca2+ buffering protein parvalbumin, they should be better able to resist the damage than the majority of motoneurons in the ALS-vulnerable motor cell groups, which lack parvalbumin (20).

ATP-Binding Cassette Transporters↗

Effectiveness of a program of early hospital discharge of cardiac surgery patients.

Managed care was the impetus for a program designed to move adult patients from acute care to the lowest level of appropriate services after cardiac surgery. Clinical pathways and a home care cardiac specialty team were the major components of the Early Discharge Program. The program was evaluated based on both financial and clinical outcomes. A convenience sample of 119 pretest patients was compared with 101 posttest patients 3 months after program implementation. Hospital length of stay decreased only 0.34 days on average, but inpatient direct variable costs decreased by an average of $1,790 per patient. Based on the 101 patients in the posttest group, $180,790 in direct variable hospital costs were saved. The largest decrease in resource use was in those patients who were discharged to home care. Complications and home caregiver burden after discharge were no higher in patients discharged early. Early discharge of cardiac surgery patients appears to be safe and cost-effective.

Aged↗

An ultrastructural double-label immunohistochemical study of the enkephalinergic input to dopaminergic neurons of the substantia nigra in pigeons.

Electron microscopic immunohistochemical double-label studies were carried out in pigeons to characterize the ultrastructural organization and postsynaptic targets of enkephalinergic (ENK+) striatonigral projection. ENK+ terminals in the substantia nigra were labeled with antileucine-enkephalin antiserum by using peroxidase-antiperoxidase methods, and dopaminergic neurons were labeled with anti-tyrosine hydroxylase antiserum by using silver-intensified immunogold methods. ENK+ terminals on dopaminergic neurons were equal in abundance to ENK+ terminals on nondopaminergic neurons, although the former were typically somewhat smaller than the latter (mean size: 0.50 vs. 0.75 micron, respectively). ENK+ terminals were evenly distributed on the cell bodies and dendrites of dopaminergic neurons, and they were evenly distributed on dendrites but rare on perikarya of nondopaminergic neurons. Transection of the basal telencephalic output revealed that 75% of the nigral ENK+ terminals were of basal telencephalic origin. These telencephalic ENK+ terminals included over 80% of those smaller than 0.80 micron on dopaminergic neurons and smaller than 1.0 micron on nondopaminergic neurons, and none greater than this in size. Both telencephalic and the nontelencephalic ENK+ nigral terminals made predominantly symmetric synapses on nigral neurons. Although the basal telencephalic ENK+ terminals uniformly targeted dendrites and perikarya, nontelencephalic ENK+ terminals seemed to avoid perikarya. The results indicate that ENK+ striatonigral neurons in birds may directly influence both dopaminergic and nondopaminergic neurons of the substantia nigra. Based on similar data for substance P-containing striatonigral terminals, the roles of enkephalin and substance P in influencing nigral dopaminergic neurons may differ slightly, as they appear to target preferentially different portions of dopaminergic neurons. The overall results in pigeons are similar to those for ENK+ terminals in the ventral tegmental area in rats, suggesting that the synaptic organization of the ENK+ input to the tegmental dopaminergic cell fields is similar in mammals and birds.

Animals↗