Effect of pituitary adenoma consistency on surgical outcomes in patients undergoing endonasal endoscopic transsphenoidal surgery.

BACKGROUND: Most pituitary adenomas (PAs) are considered to have a soft tumor consistency. However, there is a non-negligible percentage (5-13%) of tumors presenting or exhibiting a fibrous consistency that would entail a more difficult and complicated surgical excision with higher surgical morbidity and mortality rates. PURPOSE: To analyze the clinical consequences of PA tumor consistency on the surgical outcomes in patients undergoing endonasal endoscopic transsphenoidal (EET) pituitary surgery. METHODS: An ambispective study of patients with PAs operated on through an EET approach in two Spanish tertiary hospitals over the last 12 years. A total of 226 consecutive interventions were carried out in the Neurosurgery Departments of the Hospital Universitario Ramón y Cajal (HURC) and the Hospital Universitario Puerta del Sur by the same neurosurgeon. PAs were grouped into soft (n = 150) and fibrous (n = 76). All patients underwent hormonal and magnetic resonance imaging (MRI) studies before and after surgery. In addition, neurosurgical complications were recorded in each patient. RESULTS: Fibrous adenomas were independently associated with lower resection rates compared to soft adenomas (fibrous gross total resection [GTR] rate 48.7% vs. 76.3%, p < 0.001), even in those adenomas without invasion of the cavernous sinus (Knosp grades 0, I, and II). There were more intraoperative cerebrospinal fluid (CSF) leaks in patients with fibrous PAs. Moreover, fibrous PAs showed higher rates of postoperative hypopituitarism, permanent diabetes insipidus (DI) and postoperative treatments (hormonal treatment and radiotherapy). The excision of a fibrous PA required a longer surgical time (22.5 min more than soft PAs, p = 0.014), regardless of other factors. CONCLUSION: The consistency of the PAs significantly conditions both the results of surgery (lower resections rates), complications (higher incidence of postoperative hypopituitarism, permanent DI), and the prognosis (higher incidence of postoperative treatments) of the patient undergoing EET.

Screening for biosurfactant production by 2,4,6-trinitrotoluene-transforming bacteria.

AIMS: To isolate and identify TNT-transforming cultures from explosive-contaminated soils with the ability to produce biosurfactants. METHODS AND RESULTS: Bacteria (pure and mixed cultures) were selected based on their ability to transform TNT in minimum media with TNT as the sole nitrogen source and an additional carbon source. TNT-transforming bacteria were identified by 16S rRNA gene sequencing. TNT transformation rates were significantly lower when no additional carbon or nitrogen sources were added. Surfactant production was enabled by the presence of TNT. Fourteen cultures were able to transform the explosive (>50%); of these, five showed a high transformation capacity (>90%), and six produced surfactants. CONCLUSIONS: All explosive-transforming cultures contained Proteobacteria of the genera Achromobacter, Stenotrophomonas, Pseudomonas, Sphingobium, Raoultella, Rhizobium and Methylopila. These cultures transformed TNT when an additional carbon source was added. Remarkably, Achromobacter spanius S17 and Pseudomonas veronii S94 have high TNT transformation rates and are surfactant producers. SIGNIFICANCE AND IMPACT OF THE STUDY: TNT is a highly toxic, mutagenic and carcinogenic nitroaromatic explosive; therefore, bioremediation to eliminate or mitigate its presence in the environment is essential. TNT-transforming cultures that produce surfactants are a promising method for remediation. To the best of our knowledge, this is the first report that links surfactant production and TNT transformation by bacteria.

Perioperative management of patients with cardiac implantable electronic devices. / Manejo perioperatorio de los pacientes con dispositivos cardíacos electrónicos implantables.

The use of implantable cardiac devices in people of all ages is increasing, especially in the elderly population: patients with pacemakers, cardioverter-defibrillators or cardiac resynchronization therapy devices regularly present for surgery for non-cardiac causes. This review was made in order to collect and analyze the latest evidence for the proper management of implantable cardiac devices in the perioperative period. Through a detailed exploration of PubMed, Academic Search Complete (EBSCO), ClinicalKey, Cochrane (Ovid), the search software UpToDate, textbooks and patents freely available to the public on Google, we selected 33 monographs, which matched the objectives of this publication.

Pro-cognitive activity in rats of 3-furan-2-yl-N-p-tolyl-acrylamide, a positive allosteric modulator of the α7 nicotinic acetylcholine receptor.

BACKGROUND AND PURPOSE: α7 nicotinic acetylcholine receptors (α7 nAChRs) may represent useful targets for cognitive improvement. The aim of this study is to compare the pro-cognitive activity of selective α7-nAChR ligands, including the partial agonists, DMXBA and A-582941, as well as the positive allosteric modulator, 3-furan-2-yl-N-p-tolyl-acrylamide (PAM-2). EXPERIMENTAL APPROACH: The attentional set-shifting task (ASST) and the novel object recognition task (NORT) in rats, were used to evaluate the pro-cognitive activity of each ligand [i.e., PAM-2 (0.5, 1.0, and 2.0 mg·kg(-1) ), DMXBA and A-582941 (0.3 and 1.0 mg·kg(-1) )], in the absence and presence of methyllycaconitine (MLA), a selective competitive antagonist. To determine potential drug interactions, an inactive dose of PAM-2 (0.5 mg·kg(-1) ) was co-injected with inactive doses of either agonist - DMXBA: 0.1 (NORT); 0.3 mg·kg(-1) (ASST) or A-582941: 0.1 mg·kg(-1) . KEY RESULTS: PAM-2, DMXBA, and A-582941 improved cognition in a MLA-dependent manner, indicating that the observed activities are mediated by α7 nAChRs. Interestingly, the co-injection of inactive doses of PAM-2 and DMXBA or A-582941 also improved cognition, suggesting drug interactions. Moreover, PAM-2 reversed the scopolamine-induced NORT deficit. The electrophysiological results also support the view that PAM-2 potentiates the α7 nAChR currents elicited by a fixed concentration (3 µM) of DMXBA with apparent EC50 = 34 ± 3 µM and Emax = 225 ± 5 %. CONCLUSIONS AND IMPLICATIONS: Our results support the view that α7 nAChRs are involved in cognition processes and that PAM-2 is a novel promising candidate for the treatment of cognitive disorders.

3D localization of a labeled target by means of a stereo vision configuration with subvoxel resolution.

We present a method for the visual measurement of the 3D position and orientation of a moving target. Three dimensional sensing is based on stereo vision while high resolution results from a pseudo-periodic pattern (PPP) fixed onto the target. The PPP is suited for optimizing image processing that is based on phase computations. We describe experimental setup, image processing and system calibration. Resolutions reported are in the micrometer range for target position (x,y,z) and of 5:3x10(-4) rad: for target orientation (θx,θy,θz). These performances have to be appreciated with respect to the vision system used. The latter makes that every image pixel corresponds to an actual distance of 0:3x0:3 mm2 on the target while the PPP is made of elementary dots of 1 mm with a period of 2 mm. Target tilts as large as π=4 are allowed with respect to the Z axis of the system.

Interaction of benzylidene-anabaseine analogues with agonist and allosteric sites on muscle nicotinic acetylcholine receptors.

BACKGROUND AND PURPOSE: Benzylidene-anabaseines (BAs) are partial agonists of the alpha7 nicotinic acetylcholine receptor (nAChR) but their mechanism(s) of action are unknown. Our study explores several possibilities, including direct interactions of BAs with the nAChR channel. EXPERIMENTAL APPROACH: Functional and radioligand-binding assays were used to examine the interaction of two BA analogues, 3-(2,4-dimethoxybenzylidene)-anabaseine (DMXBA) and its primary metabolite 3-(4-hydroxy-2-methoxybenzylidene)-anabaseine (4OH-DMXBA) with both agonist and non-competitive antagonist (NCA)-binding sites on muscle-type nAChRs. KEY RESULTS: Both BAs non-competitively inhibited ACh activation of human fetal muscle nAChRs and sterically inhibited the specific binding of the NCAs [piperidyl-3,4-3H(N)]-(N-(1-(2-thienyl)cyclohexyl)-3,4-piperidine ([(3)H]TCP) and [(3)H]dizocilpine to Torpedo nAChRs in the desensitized state. These compounds modulated [(3)H]tetracaine, [(14)C]amobarbital and [(3)H]TCP binding to resting nAChRs by allosteric mechanisms. Both BAs enhanced [(3)H]TCP binding when the nAChR was initially in the resting but activatable state, suggesting that both compounds desensitized the Torpedo nAChR. Although DMXBA failed to activate human fetal muscle nAChRs, 4OH-DMXBA was found to be a partial agonist. [(3)H]Nicotine competition-binding experiments confirmed that 4OH-DMXBA has higher affinity than DMXBA for the agonist sites, and that DMXBA is also a competitive antagonist. CONCLUSIONS AND IMPLICATIONS: 3-(4-hydroxy-2-methoxybenzylidene)-anabaseine is a partial agonist for human fetal muscle nAChRs, whereas DMXBA only has competitive and NCA activities. The NCA-binding site for BAs overlaps both the phencyclidine- and dizocilpine-binding sites in the desensitized Torpedo nAChR ion channel. The desensitizing property of BAs suggests another possible mode of non-competitive inhibition in addition to direct channel-blocking mechanisms.

Caso clínico: aspergilosis pulmonar invasiva tratada con Voriconazol en paciente con Síndrome de Inmunodeficiencia Adquirida / Clinical case: invasive pulmonary aspergillosis treated with Voriconazol in an AIDS patient

Se reporta un caso clínico de aspergilosis pulmonar invasiva en un paciente de 29 años VIH(+) en etapa SIDA, sin antecedentes mórbidos conocidos, con diagnóstico inicial de neumonía por Pneumocystis jirovecii. Fue tratado con éxito, pero sin asistir a controles posterior a su alta . Tres meses después ingresa al servicio de Urgencias del Hospital Gustavo Fricke con tos productiva mucopurulenta, disnea progresiva, fiebre intermitente y compromiso del estado general. La radiografía de tórax sugirió neumonía atípica, detectándose en los exámenes Pneumocystis jirovecii y Enterobacter aerógenes , por lo que se inicia tratamiento con Cotrimoxazol y Ertapenem. En los cultivos en agar Sabouraud se detectó abundante desarrollo de Aspergillus fumigatus , por lo que se empieza tratamiento con anfotericina B en dosis crecientes hasta alcanzar 50 mg/día, sin embargo, por reacciones adversas severas se decidió tratamiento con Voriconazol intravenoso y luego oral, con buena respuesta clínica, radiológica y de laboratorio. Es dado de alta con tratamiento con Voriconazol oral, además de profilaxis secundaria para P. jirovecii y Mycobaterium avium.

A clinical case of an invasive pulmonary aspergillosis in a 29 aged VIH (+) patient, at an AIDS stage, lacking any known morbid data, and bearing an initial diagnosis of pneumonia by Pneumocystis jirovecii is herein described. Was successfully treated even though he failed to attend subsequent health controls. Three months later he is admitted in the Hospital Gustavo Fricke, showing productive mucupurulent cough, progressive disnea, intermittent fever and his overall health condition resulting deeply compromised. Thorax X-ray revealed an atypical pneumonia together with the presence of P. jirovecii and Enterobacter aerogenes, and decided to treat him with Cotrimoxazol and Ertapenem. Meanwhile in agar cultures a heavy development of Aspergillus fumigatus was detected, thus the patient was given Anfotericina B in increasing doses up to reach 50mg/day; however due to some severe adverse reactions, the treatment with intravenous and later oral Voriconazol, which rendered satisfactory clinical, radiological and laboratory responses was ultimately preferred. The patient is discharged from the hospital and advised to continue with oral Voriconazol besides undergoing secondary profilaxis for P. jirovecii and Mycobaterium avium.

Heavy metal content in soils under different wastewater irrigation patterns in Chihuahua, Mexico.

An area near the city of Chihuahua has been traditionally irrigated with wastewater to grow forage crops. It has been hypothesized that metal levels could be found in these soils high enough to cause potential health problems to the population. The objective of this study was to determine heavy metal concentrations in different soils due to irrigation practices. Four soil types were evaluated; a soil with a past and present history of wastewater irrigation (S1), a soil with a history of wastewater irrigation until 2003 (S2), a soil with no irrigation history (S3), and a soil similar to S1 and adjacent to the river where the wastewater is transported (S11). Three soil depths were evaluated; 0-15, 15-30 and 30-50 cm. Consequently, a total of 150 soil samples were analyzed evaluating pH, EC, OM and the following elements; Na, K, Cd, Pb, Ni, Cr, Cu and Fe. The pH (P=0.000) and EC (P=0.000) were different for each soil type but no differences were noted for soil depth and the interaction. Maximum pH levels were noted in S3 with a value of 8.74 while maximum EC was observed in S1 with a value of 0.850 dSm-1. The OM level was different for soil type (P=0.000), soil depth (P=0.005) and the interaction (P=0.014). S1 and S11 obtained maximum levels of OM while minimum levels were noted in S3. Maximum OM levels were observed at the 0-15 cm depth followed by the 15-30 cm depth and finally at the 30-50 cm depth. The highest concentration of metals was as follows: K in S1 (359.3 mg kg-1); Cd in S1 (4.48 mg kg-1); Pb in S11 (155.83 mg kg-1); Ni in S1 (10.74 mg kg-1); Cu in S1 (51.36 mg kg-1); B in S3 (41.5 mg kg-1); Fe in S3 (20,313.0 mg kg-1), Cr in S3 (44.26 mg kg-1) and Na in S3 (203.0 mg kg-1). The conclusion is that some metals are present in the soils due to anthropogenic activities but others are present in natural forms.

Effects of alpha-tocopherol on oxidative status and metabolic profile in overweight women.

Despite extensive research, the effects of alpha-tocopherol supplementation remain controversial. Few studies have been focused on obese and overweight people. We examined the effects of alpha-tocopherol (AT) on the oxidative status and metabolic profile in overweight women. Sixteen overweight women between the ages of 40-60 years old, received AT, 800 IU/day during 12 weeks, followed by a 6-week washout period. Blood samples were taken at the beginning and then every 6 weeks until the end of the study. AT, retinol, malondialdehyde (MDA), total antioxidant status (TAS), selenium-dependent glutathione peroxidase (GPx) and CuZn-superoxide dismutase (SOD) were quantified to evaluate the oxidative stress. The metabolic profile was estimated by measuring glycated hemoglobin (HbA1c) in erythrocytes and glucose, phosphate, magnesium, lipid and lipoprotein concentrations in serum. Under AT administration HbA1c, serum- MDA levels and erythrocyte GPx activity were markedly reduced. TAS, AT and Mg2+ concentrations in serum and SOD activity in erythrocytes were higher after AT treatment. Body weight; glucose, lipid and retinol concentrations, or blood cells count were unchanged. Lipid peroxidation was considerably reduced in AT treated women and also improved serum antioxidant status was observed, but the imbalanced response between erythrocyte SOD and GPx activities could affect normal response to oxidative stress.

Lipid-protein interactions and effect of local anesthetics in acetylcholine receptor-rich membranes from Torpedo marmorata electric organ.

The selectivity of lipid-protein interaction for spin-labeled phospholipids and gangliosides in nicotinic acetylcholine receptor-rich membranes from Torpedo marmorata has been studied by ESR spectroscopy. The association constants of the spin-labeled lipids (relative to phosphatidylcholine) at pH 8.0 are in the order cardiolipin (5.1) approximately equal to stearic acid (4.9) approximately equal to phosphatidylinositol (4.7) > phosphatidylserine (2.7) > phosphatidylglycerol (1.7) > G(D1b) approximately equal to G(M1) approximately equal to G(M2) approximately equal to G(M3) approximately equal to phosphatidylcholine (1.0) > phosphatidylethanolamine (0.5). No selectivity for mono- or disialogangliosides is found over that for phosphatidylcholine. Aminated local anesthetics were found to compete with spin-labeled phosphatidylinositol, but to a much lesser extent with spin-labeled stearic acid, for sites on the intramembranous surface of the protein. The relative association constant of phosphatidylinositol was reduced in the presence of the different local anesthetics to the following extents: tetracaine (55%) > procaine (35%) approximately benzocaine (30%). For stearic acid, only tetracaine gave an appreciable reduction (30%) in association constant. These displacements represent an intrinsic difference in affinity of the local anesthetics for the lipid-protein interface because the membrane partition coefficients are in the order benzocaine >> tetracaine approximately procaine.

Interaction of barbiturate analogs with the Torpedo californica nicotinic acetylcholine receptor ion channel.

Barbiturate-induced anesthesia is a complex mechanism that probably involves several ligand-gated ion channel superfamilies. One of these superfamilies includes the archetypical nicotinic acetylcholine receptor (nAChR), in which barbiturates act as noncompetitive antagonists. In this regard, we used the Torpedo californica nAChR and a series of barbiturate analogs to characterize the barbiturate binding site(s) on this superfamily member. [(14)C]Amobarbital binds to one high-affinity (K(d) = 3.7 microM) and several (approximately 11) low-affinity (K(d) = 930 microM) sites on the resting and desensitized nAChRs, respectively. Characteristics of the barbiturate binding site on the resting nAChR include: (1) a tight structure-activity relationship. For example, the barbiturate isobarbital [5-ethyl-5'-(2-methylbutyl) barbituric acid] is >10-fold less potent than its formula isomer amobarbital [5-ethyl-5'-(3-methylbutyl) barbituric acid] in inhibiting [(14)C]amobarbital binding. (2) A binding locus within the pore of the nAChR ion channel. Each of the barbiturate analogs inhibited the binding of [(3)H]tetracaine or photoincorporation of 3-trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine in a mutually exclusive manner. (3) Stereoselective binding. The R(+)-enantiomers of isobarbital and pentobarbital are approximately 2-fold more potent in inhibiting 3-trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine photoincorporation than the S(-)-enantiomers. Finally, molecular modeling suggests that within the channel, the pyrimidine ring of the barbiturate is located just above the highly conserved leucine ring (M2--9; e.g., delta Leu-265), whereas the 5' side chain projects downward, and depending upon its conformation, introduces steric hindrance to binding because of the restriction in the lumen of the channel introduced by the leucine side chains.

Characterization of the dizocilpine binding site on the nicotinic acetylcholine receptor.

Although the dissociative anesthetic dizocilpine [(+)-MK-801] inhibits nicotinic acetylcholine receptor (AChR) function in a noncompetitive manner, the location of the dizocilpine binding site(s) has yet to be clearly established. Thus, to characterize the binding site for dizocilpine on the AChR we examined 1) the dissociation constant (K(d)) and stoichiometry of [(3)H]dizocilpine binding; 2) the displacement of dizocilpine radioligand binding by noncompetitive inhibitors (NCIs) and conversely dizocilpine displacement of fluorescent and radiolabeled NCIs from their respective high-affinity binding sites on the AChR; and 3) photoaffinity labeling of the AChR using (125)I-dizocilpine. The results establish that one high-affinity (K(d) = 4.8 microM) and several (3-6) low-affinity (K(d) = approximately 140 microM) binding sites exist for dizocilpine on the desensitized and resting AChR, respectively. The binding of the fluorescent NCIs ethidium, quinacrine, and crystal violet as well as [(3)H]thienylcyclohexylpiperidine was inhibited by dizocilpine on desensitized AChRs. However, Schild-type analyses indicate that only the inhibition of quinacrine in the desensitized state seems to be mediated by a mutually exclusive action. Photoaffinity labeling of the AChR by (125)I-dizocilpine was primarily restricted to the alpha1 subunit and subsequent mapping revealed that the principal sites of labeling are localized to the M4 (approximately 70%) and M1 (30%) transmembrane domains. Collectively, the data indicate that the high-affinity dizocilpine binding site is not located in the lumen of the ion channel but probably near the quinacrine binding locus at a nonluminal domain in the AChR desensitized state.

Alpha-conotoxins.

alpha-Conotoxins (alpha-CgTxs) are a family of Cys-enriched peptides found in several marine snails from the genus Conus. These small peptides behave pharmacologically as competitive antagonists of the nicotinic acetylcholine receptor (AChR). The data indicate that (1) alpha-CgTxs are able to discriminate between muscle- and neuronal-type AChRs and even among distinct AChR subtypes; (2) the binding sites for alpha-CgTxs are located, like other cholinergic ligands, at the interface of alpha and non-alpha subunits (gamma, delta, and epsilon for the muscle-type AChR, and beta for several neuronal-type AChRs); (3) some alpha-CgTxs differentiate the high- from the low-affinity binding site found on either alpha/non-alpha subunit interface; and that (4) specific residues in the cholinergic binding site are energetically coupled with their corresponding pairs in the toxin stabilizing the alpha-CgTx-AChR complex. The alpha-CgTxs have proven to be excellent probes for studying the structure and function of the AChR family.

Localization of agonist and competitive antagonist binding sites on nicotinic acetylcholine receptors.

Identification of all residues involved in the recognition and binding of cholinergic ligands (e.g. agonists, competitive antagonists, and noncompetitive agonists) is a primary objective to understand which structural components are related to the physiological function of the nicotinic acetylcholine receptor (AChR). The picture for the localization of the agonist/competitive antagonist binding sites is now clearer in the light of newer and better experimental evidence. These sites are located mainly on both alpha subunits in a pocket approximately 30-35 A above the surface membrane. Since both alpha subunits are identical, the observed high and low affinity for different ligands on the receptor is conditioned by the interaction of the alpha subunit with other non-alpha subunits. This molecular interaction takes place at the interface formed by the different subunits. For example, the high-affinity acetylcholine (ACh) binding site of the muscle-type AChR is located on the alphadelta subunit interface, whereas the low-affinity ACh binding site is located on the alphagamma subunit interface. Regarding homomeric AChRs (e.g. alpha7, alpha8, and alpha9), up to five binding sites may be located on the alphaalpha subunit interfaces. From the point of view of subunit arrangement, the gamma subunit is in between both alpha subunits and the delta subunit follows the alpha aligned in a clockwise manner from the gamma. Although some competitive antagonists such as lophotoxin and alpha-bungarotoxin bind to the same high- and low-affinity sites as ACh, other cholinergic drugs may bind with opposite specificity. For instance, the location of the high- and the low-affinity binding site for curare-related drugs as well as for agonists such as the alkaloid nicotine and the potent analgesic epibatidine (only when the AChR is in the desensitized state) is determined by the alphagamma and the alphadelta subunit interface, respectively. The case of alpha-conotoxins (alpha-CoTxs) is unique since each alpha-CoTx from different species is recognized by a specific AChR type. In addition, the specificity of alpha-CoTxs for each subunit interface is species-dependent. In general terms we may state that both alpha subunits carry the principal component for the agonist/competitive antagonist binding sites, whereas the non-alpha subunits bear the complementary component. Concerning homomeric AChRs, both the principal and the complementary component exist on the alpha subunit. The principal component on the muscle-type AChR involves three loops-forming binding domains (loops A-C). Loop A (from mouse sequence) is mainly formed by residue Y(93), loop B is molded by amino acids W(149), Y(152), and probably G(153), while loop C is shaped by residues Y(190), C(192), C(193), and Y(198). The complementary component corresponding to each non-alpha subunit probably contributes with at least four loops. More specifically, the loops at the gamma subunit are: loop D which is formed by residue K(34), loop E that is designed by W(55) and E(57), loop F which is built by a stretch of amino acids comprising L(109), S(111), C(115), I(116), and Y(117), and finally loop G that is shaped by F(172) and by the negatively-charged amino acids D(174) and E(183). The complementary component on the delta subunit, which corresponds to the high-affinity ACh binding site, is formed by homologous loops. Regarding alpha-neurotoxins, several snake and alpha-CoTxs bear specific residues that are energetically coupled with their corresponding pairs on the AChR binding site. The principal component for snake alpha-neurotoxins is located on the residue sequence alpha1W(184)-D(200), which includes loop C. In addition, amino acid sequence 55-74 from the alpha1 subunit (which includes loop E), and residues gammaL(119) (close to loop F) and gammaE(176) (close to loop G) at the low-affinity binding site, or deltaL(121) (close to the homologous region of loop G) at the high-affinity binding site, are i

[Tissue infections caused by group A beta hemolytic Streptococcus in a regional hospital of Talca, Chile]. / Infecciones tisulares por Streptococcus beta-hemolítico grupo A en Hospital Regional de Talca.

BACKGROUND: A resurgence of group A beta hemolytic Streptococcus infections such as fasciitis, cellulitis and Streptococcal Toxic Syndrome has been observed recently. AIM: To study the clinical features of patients with group A beta hemolytic Streptococcus infections in a regional hospital. PATIENTS AND METHODS: Retrospective review of medical records of patients notified as having a group A beta hemolytic Streptococcus tissue infection, between 1994 and 1999. RESULTS: Twenty four patients were notified in the period as having a group A beta hemolytic Streptococcus infection and 18 (13 male, mean age 39 tears old) had tissue involvement. Eleven patients had a fasciitis (61%), six had a cellulitis (33%) and 14 patients (77%), a Streptococcal Toxic Shock Syndrome. Eight patients died during hospital stay. The infection portal of entry was identify in 13 patients (the skin in 10 and intramuscular injections in three). Decreased patients had a longer lapse of disease before admission than patients discharged alive (5(range 3h-7 days) and 2.1 (range 6h-5 days) respectively). In seven patients a quick serological test, designed for pharyngeal infections was performed and it was positive in five. Blood cultures were positive in seven patients and in 11, the germ was isolated from the lesions. CONCLUSIONS: As the early diagnosis of group A beta hemolytic Streptococcus tissue infections has a prognostic value, the population should be instructed to recognize early signs and symptoms of these infections.

Role of local anesthetics on both cholinergic and serotonergic ionotropic receptors.

A great body of experimental evidence indicates that the main target for the pharmacological action of local anesthetics (LAs) is the voltage-gated Na+ channel. However, the epidural and spinal anesthesia as well as the behavioral effects of LAs cannot be explained exclusively by its inhibitory effect on the voltage-gated Na+ channel. Thus, the involvement of other ion channel receptors has been suggested. Particularly, two members of the neurotransmitter-gated ion channel receptor superfamily, the nicotinic acetylcholine receptor (AChR) and the 5-hydroxytryptamine receptor (5-HT3R type). In this regard, the aim of this review is to explain and delineate the mechanism by which LAs inhibit both ionotropic receptors from peripheral and central nervous systems. Local anesthetics inhibit the ion channel activity of both muscle- and neuronal-type AChRs in a noncompetitive fashion. Additionally, LAs inhibit the 5-HT3R by competing with the serotonergic agonist binding sites. The noncompetitive inhibitory action of LAs on the AChR is ascribed to two possible blocking mechanisms. An open-channel-blocking mechanism where the drug binds to the open channel and/or an allosteric mechanism where LAs bind to closed channels. The open-channel-blocking mechanism is in accord with the existence of high-affinity LA binding sites located in the ion channel. The allosteric mechanism seems to be physiologically more relevant than the open-channel-blocking mechanism. The inhibitory property of LAs is also elicited by binding to several low-affinity sites positioned at the lipid-AChR interface. However, there is no clearcut evidence indicating whether these sites are located at either the annular or the nonannular lipid domain. Both tertiary (protonated) and quaternary LAs gain the interior of the channel through the hydrophilic pathway formed by the extracellular ion channel's mouth with the concomitant ion flux blockade. Nevertheless, an alternative mode of action is proposed for both deprotonated tertiary and permanently-uncharged LAs: they may pass from the lipid membrane core to the lumen of the ion channel through a hydrophobic pathway. Perhaps this hydrophobic pathway is structurally related to the nonannular lipid domain. Regarding the LA binding site location on the 5-HT3R, at least two amino acids have been involved. Glutamic acid at position 106 which is located in a residue sequence homologous to loop A from the principal component of the binding site for cholinergic agonists and competitive antagonists, and Trp67 which is positioned in a stretch of amino acids homologous to loop F from the complementary component of the cholinergic ligand binding site.

5-Doxylstearate-induced displacement of phencyclidine from its low-affinity binding sites on the nicotinic acetylcholine receptor.

Fatty acids as well as phencyclidine (PCP) inhibit the ion channel activity of the nicotinic acetylcholine receptor (AChR) by a noncompetitive mechanism. However, the exact localization of the fatty acid binding sites is unknown and, thus, the noncompetitive inhibitory mechanism for these endogenous modulators remains to be elucidated. In an attempt to determine the location of the fatty acid binding sites, we study the mutually exclusive action between 5-doxylstearate (5-SASL), a derivative of the endogenous noncompetitive antagonist (NCA) stearic acid, and other exogenous NCAs. For this purpose, both equilibrium and competitive binding assays using fluorescent and radiolabeled ligands were performed on desensitized AChRs. More specifically, we determined: (i) the effect of 5-SASL on the binding of the exogenous NCA [(3)H]PCP; (ii) the effect of 5-SASL on the binding of either quinacrine or ethidium, two fluorescent NCAs from exogenous origin; and (iii) the PCP-induced displacement of quinacrine and ethidium from their respective high-affinity binding sites. Our first target (i) is carried out by measuring the [(3)H]PCP binding in the absence or in the presence of increasing concentrations of 5-SASL. We found that 5-SASL displaces PCP from its low-affinity binding sites. The low-affinity PCP binding sites were pharmacologically characterized by an apparent dissociation constant (K(d)) of 6.1 +/- 5.0 microM and a stoichiometry of 3.7 +/- 1.5 sites per AChR. The fact that 5-SASL increased the apparent K(d) without changing the number of sites per AChR is indicative of a mutually exclusive action. From these results, an apparent inhibition constant (K(i)) of 75 +/- 31 microM for 5-SASL was calculated. In addition, 5-SASL affected neither the apparent K(d) (0.46 +/- 0.37 microM) nor the stoichiometry (1.07 +/- 0.57 sites per AChR) of the high-affinity PCP binding site. The second objective (ii) is achieved by titrating either quinacrine or ethidium into AChR native membranes in the absence or in the presence of increasing concentrations of 5-SASL. These experiments showed that 5-SASL efficiently increased the apparent K(d) of quinacrine without perturbing the interaction of ethidium with its high-affinity locus. Considering that (a) 5-SASL effectively quenched the AChR-bound quinacrine fluorescence (H. R. Arias, Biochim. Biophys. Acta 1347, 9-22, 1997) and (b) fluorescence-quenching is a short-range process, it is possible to suggest that 5-SASL displaces quinacrine from its high-affinity binding site by a steric mechanism. In this regard, a K(i) of 38 +/- 5 microM for 5-SASL was calculated. Concerning the last objective (iii), AChR-bound quinacrine or ethidium was back titrated with PCP. Two PCP K(i) values were obtained by fitting the displacement plots by nonlinear regression with two components. The lowest K(i) values obtained for either quinacrine (0.86 +/- 0.37 microM) or ethidium (0. 29 +/- 0.23 microM) displacement from their respective high-affinity binding sites coincide with the previously determined high-affinity [(3)H]PCP K(d). In addition, the highest K(i) values obtained for either NCA displacement are in the same concentration range as the observed low-affinity [(3)H]PCP K(d). Taking into account all experimental data, we reached the following conclusions: (i) fatty acid molecules, or at least 5-SASL, sterically interact with both the PCP low-affinity and the quinacrine high-affinity binding sites; (ii) the low-affinity PCP binding sites, as well as the high-affinity quinacrine locus, are located at the nonannular lipid domain of the AChR; and, finally, (iii) fatty acid molecules are not accessible to the lumen of the ion channel, indicating an allosteric mode of action for fatty acids to inhibit ion flux. Thus, the 5-SASL, the quinacrine high-affinity, and the PCP low-affinity binding sites are all located at overlapping nonannular loci on the muscle-type AChR.

Binding sites for exogenous and endogenous non-competitive inhibitors of the nicotinic acetylcholine receptor.

The nicotinic acetylcholine receptor (AChR) is the paradigm of the neurotransmitter-gated ion channel superfamily. The pharmacological behavior of the AChR can be described as three basic processes that progress sequentially. First, the neurotransmitter acetylcholine (ACh) binds the receptor. Next, the intrinsically coupled ion channel opens upon ACh binding with subsequent ion flux activity. Finally, the AChR becomes desensitized, a process where the ion channel becomes closed in the prolonged presence of ACh. The existing equilibrium among these physiologically relevant processes can be perturbed by the pharmacological action of different drugs. In particular, non-competitive inhibitors (NCIs) inhibit the ion flux and enhance the desensitization rate of the AChR. The action of NCIs was studied using several drugs of exogenous origin. These include compounds such as chlorpromazine (CPZ), triphenylmethylphosphonium (TPMP+), the local anesthetics QX-222 and meproadifen, trifluoromethyl-iodophenyldiazirine (TID), phencyclidine (PCP), histrionicotoxin (HTX), quinacrine, and ethidium. In order to understand the mechanism by which NCIs exert their pharmacological properties several laboratories have studied the structural characteristics of their binding sites, including their respective locations on the receptor. One of the main objectives of this review is to discuss all available experimental evidence regarding the specific localization of the binding sites for exogenous NCIs. For example, it is known that the so-called luminal NCIs bind to a series of ring-forming amino acids in the ion channel. Particularly CPZ, TPMP+, QX-222, cembranoids, and PCP bind to the serine, the threonine, and the leucine ring, whereas TID and meproadifen bind to the valine and extracellular rings, respectively. On the other hand, quinacrine and ethidium, termed non-luminal NCIs, bind to sites outside the channel lumen. Specifically, quinacrine binds to a non-annular lipid domain located approximately 7 A from the lipid-water interface and ethidium binds to the vestibule of the AChR in a site located approximately 46 A away from the membrane surface and equidistant from both ACh binding sites. The non-annular lipid domain has been suggested to be located at the intermolecular interfaces of the five AChR subunits and/or at the interstices of the four (M1-M4) transmembrane domains. One of the most important concepts in neurochemistry is that receptor proteins can be modulated by endogenous substances other than their specific agonists. Among membrane-embedded receptors, the AChR is one of the best examples of this behavior. In this regard, the AChR is non-competitively modulated by diverse molecules such as lipids (fatty acids and steroids), the neuropeptide substance P, and the neurotransmitter 5-hydroxytryptamine (5-HT). It is important to take into account that the above mentioned modulation is produced through a direct binding of these endogenous molecules to the AChR. Since this is a physiologically relevant issue, it is useful to elucidate the structural components of the binding site for each endogenous NCI. In this regard, another important aim of this work is to review all available information related to the specific localization of the binding sites for endogenous NCIs. For example, it is known that both neurotransmitters substance P and 5-HT bind to the lumen of the ion channel. Particularly, the locus for substance P is found in the deltaM2 domain, whereas the binding site for 5-HT and related compounds is putatively located on both the serine and the threonine ring. Instead, fatty acid and steroid molecules bind to non-luminal sites. More specifically, fatty acids may bind to the belt surrounding the intramembranous perimeter of the AChR, namely the annular lipid domain, and/or to the high-affinity quinacrine site which is located at a non-annular lipid domain. Additionally, steroids may bind to a site located on the extracellular hydrophi

[Gene therapy and cancer]. / Terapia génica y cáncer.

Worldwide, cancer remains one of the major causes of mortality. For example, American epidemiologists have projected that for 1996 nearly 550,000 USA patients will die-up from 310,000 in 1970. Furthermore, some 40 percent of Americans will eventually be stricken with the disease, and more than one in five will die of it. While this trend remains the same for developed countries, the situation is worse in developing countries. Although with chemotherapy, radiotherapy and surgery it is often possible to reduce the tumor burden dramatically, regrowth from residual cells is depressingly common, and the disease eventually becomes resistant to all standard forms of therapy, which is reflected by the fact that the survival rate has not improved during the last 20 years. Hence, it is hoped that new, simple, sensitive and specific methodologies will help to reduce the cancer incidence and, within this context, gene therapy has been heralded for its potential to revolutionize modern medicine through the use of recombinant DNA technology, in the search for cancer treatment solutions. A review of the different strategies employed in gene therapy and the analysis of more promising areas of development in gene therapy is presented.

Noncompetitive inhibition of nicotinic acetylcholine receptors by endogenous molecules.

Over the past few decades much effort has been expended elucidating the key domains of the nicotinic acetylcholine receptor (AChR) responsible for agonist binding, ion conduction, and gating. An emerging concept in the receptor field has been to consider the receptor entity as a signal transducer that suffers modulatory control by allosterically acting ligands. Of particular interest are the molecules that inhibit the agonist-evoked ion flux activity in a noncompetitive manner: the so-called noncompetitive inhibitors (NCIs). The actual knowledge on the action of NCIs was obtained by using several drugs from exogenous origin. However, several lines of investigation indicate that the receptor protein can be modulated by endogenous substances other than acetylcholine. In this regard, we outline the progress evidenced on the localization of binding sites for drugs of endogenous origin that have been found to directly interact with the AChR in a noncompetitive fashion. Among them we can quote lipids such as steroids and fatty acids, the neurotransmitter 5-hydroxytryptamine (5-HT) and related compounds, as well as the neuropeptide substance P. We present the available experimental evidence indicating the existence of both luminal (located into the ion channel) and nonluminal (located out of the ion channel) binding sites for endogenous NCIs. Particularly, the binding site for substance P is found in the delta M2 domain. In addition, the locus for 5-HT is putatively located in the ion channel close to the serine ring, whereas the binding site for two competitive antagonists of 5-HT receptors (e.g., methysergide and spiperone) is located closer to the external end of the ion channel. Instead, fatty acid and steroid molecules bind to nonluminal sites. More specifically, fatty acids may bind to the annular lipid domain of the AChR or/and to the high-affinity quinacrine site (a NCI from exogenous origin) which is located at a nonannular lipid domain. Additionally, steroids may bind to a site located on the extracellular hydrophilic domain of the AChR or/and at the lipid-protein interface, specifically, at the annular lipid domain and/or close to the nonannular quinacrine binding site.