Efficacy and feasibility of basal-bolus insulin regimens and a discharge-strategy in hospitalised patients with type 2 diabetes--the HOSMIDIA study.

AIMS: Guidelines recommend use of basal-bolus insulin in hospitalised patients with hyperglycaemia, but information about implementation and medication reconciliation at discharge is scarce. The HOSMIDIA study evaluated a management program involving basal-bolus insulin and an algorithm for medication reconciliation at discharge in non-critically ill hospitalised patients with type 2 diabetes in clinical practice. METHODS: HOSMIDIA was a prospective, observational study performed during routine clinical practice at 15 Spanish hospitals during hospitalisation, with follow-up 3 months postdischarge. Study patients (n = 134) received a basal-bolus regimen with insulin glargine during hospitalisation and treatment at discharge was adjusted according to a simple algorithm. The control group (n = 62) included patients with similar characteristics hospitalised during the month before study initiation and had no follow-up after discharge. RESULTS: Compared with control subjects, patients in the prospective study achieved lower mean total (167.7 ± 41.1 vs. 190.5 ± 53.3 mg/dl) preprandial (164.2 ± 42.4 vs. 189.6 ± 52.6 mg/dl; p < 0.001) and fasting (137.0 ± 42.2 vs. 165.8 ± 56.5 mg/dl) blood glucose levels while hospitalised, without increased hypoglycaemic episodes (17.7% vs. 19.3% patients). In the prospective study, glycaemic control improved from admission to discharge, with control maintained 3 months after discharge. The main treatment modification at discharge compared with admission was addition of basal insulin, and treatment at discharge was maintained at 3 months in 89% of patients. CONCLUSION: The HOSMIDIA study confirmed that management of hyperglycaemia with basal-bolus insulin is feasible and effective in routine clinical practice, and that a simple strategy facilitating the reconciliation of medication on discharge can improve glycaemic control postdischarge.

[Placenta percreta with bladder invasion: an uncommon cause of hematuria during pregnancy]. / Placenta percreta con invasión vesical: una causa infrecuente de hematuria en la embarazada.

Placenta percreta with bladder invasion is a very uncommon condition that can lead to very severe complications in pregnant women. Although it is often diagnosed during delivery, imaging techniques are very useful for early diagnosis, which is fundamental for planning surgery and avoiding potentially lethal complications. We present the case of a woman with a history of cesarean section who presented with hematuria and low back pain. The diagnosis of placenta percreta with bladder invasion was suggested after ultrasonography and magnetic resonance imaging and was confirmed at surgery. We provide a brief review of the literature, emphasizing the role of imaging techniques.

Stability of graph communities across time scales.

The complexity of biological, social, and engineering networks makes it desirable to find natural partitions into clusters (or communities) that can provide insight into the structure of the overall system and even act as simplified functional descriptions. Although methods for community detection abound, there is a lack of consensus on how to quantify and rank the quality of partitions. We introduce here the stability of a partition, a measure of its quality as a community structure based on the clustered autocovariance of a dynamic Markov process taking place on the network. Because the stability has an intrinsic dependence on time scales of the graph, it allows us to compare and rank partitions at each time and also to establish the time spans over which partitions are optimal. Hence the Markov time acts effectively as an intrinsic resolution parameter that establishes a hierarchy of increasingly coarser communities. Our dynamical definition provides a unifying framework for several standard partitioning measures: modularity and normalized cut size can be interpreted as one-step time measures, whereas Fiedler's spectral clustering emerges at long times. We apply our method to characterize the relevance of partitions over time for constructive and real networks, including hierarchical graphs and social networks, and use it to obtain reduced descriptions for atomic-level protein structures over different time scales.

[Unicompartmental knee prosthesis]. / Prótesis unicompartimental de rodilla.

In a patient with severe unicompartmental knee osteoarthritis where conservative treatments have been exhausted, with painful symptoms located on the affected side and with a reducible axis, the unicompartmental knee prosthesis (UKP) is the first option for our work group. Within the study to confirm the diagnosis and plan the surgery, weight-bearing knee x-rays, Rosenberg x-rays, and teleradiographs of the lower extremities stand out. The objective of surgery is to replace the affected area, restoring the anatomy with an adequate balance of soft tissues. Regarding alignment, the challenge is not to overload the opposite side or that of the prosthesis. There are mobile and fixed plates and although the clinical and survival results are similar, in recent years with the incorporation of robotic surgery, the balance has tipped towards the use of fixed plates. The clinical and functional results are better and there are fewer complications than when total knee prostheses (TKP) are used in the same type of patients. The survival studied in registries is lower than for TKP, but when used in high-flow centers where the percentage of UKP is close to a third of the total with strict patient selection, the duration is as good as in PTR.

En un paciente con artrosis unicompartimental de rodilla grave, donde se agotaron los tratamientos conservadores que tiene clínica dolorosa localizada en el lado afectado y con alteración del eje reductible, la prótesis unicompartimenal de rodilla (PUR) es la primera opción para nuestro grupo de trabajo. Dentro del estudio para confirmar el diagnóstico y planificar la cirugía destacan las radiografías de rodillas con carga, radiografía de Rosenberg y telerradiografías de extremidades inferiores. El objetivo de la cirugía es reemplazar la zona afectada, restituyendo la anatomía con un adecuado balance de partes blandas. Respecto al alineamiento el desafío es no sobrecargar el lado contrario ni tampoco el de la prótesis. Existen de platillo móvil y fijo y aunque los resultados clínicos y de supervivencia son semejantes, en los últimos años con la incorporación de la cirugía robótica, la balanza se ha inclinado para el uso de los platillos fijos. En pacientes con artrosis unicompartimental los resultados clínicos y funcionales son mejores con PUR y tiene menos complicaciones que cuando se usan prótesis totales (PTR). La supervivencia estudiada en registros es menor que para PTR, pero cuando se usa en centros de alto flujo en que el porcentaje de PUR es cercano a un tercio del total con una estricta selección de pacientes, la duración es tan buena como en la PTR.

Usefulness of Ultrasound in Assessing the Impact of Bariatric Surgery on Body Composition: a Pilot Study.

BACKGROUND: Bariatric surgery (BS) has a significant impact on body composition. The purpose of the study is to evaluate the usefulness of musculoskeletal ultrasound (MUS) to bioelectrical impedance (BIA) in the follow-up of patients undergoing BS in terms of body composition and quality of life (QoL). METHODS: This is a prospective pilot study including 32 subjects (75% female, mean age: 49.15 ± 1.9 years) who underwent BS. Fat mass (FM), lean mass (LM), and skeletal muscle index (SMI) were calculated by BIA. MUS measured subcutaneous fat (SF) and thigh muscle thickness (TMT) of the quadriceps. QoL was assessed by the Moorehead-Ardelt questionnaire. All these measurements were performed 1 month prior to BS and at 12-month follow-up. RESULTS: The mean BMI decreased by 6.63 ± 1.25 kg/m2 (p=0.001). We observed significant reductions in FM (p=0.001) and SF (p=0.007) and in LM (p=0.001) but not in SMI and TMT. We found a correlation between the FM and SF (pre-surgical, r=0.42, p=0.01; post-surgical, r=0.52, p=0.003) and between SMI and TMT (pre-surgical, r=0.35, p=0.04; post-surgical, r=0.38, p=0.03). QoL test showed significant improvement (p=0.001). In addition, a correlation between the QoL questionnaire and TMT post-surgery (r=0.91, p=0.019) was observed. However, we did not find any statistically significant correlation between QoL assessment and SMI or LM. CONCLUSIONS: Our results suggest that MUS can be complementary to BIA for the evaluation and the follow-up of body composition after BS. TMT of quadriceps can provide relevant information about regional sarcopenia and has a significant correlation with QoL.

GLUT2 expression by glial fibrillary acidic protein-positive tanycytes is required for promoting feeding-response to fasting.

Feeding behavior is a complex process that depends on the ability of the brain to integrate hormonal and nutritional signals, such as glucose. One glucosensing mechanism relies on the glucose transporter 2 (GLUT2) in the hypothalamus, especially in radial glia-like cells called tanycytes. Here, we analyzed whether a GLUT2-dependent glucosensing mechanism is required for the normal regulation of feeding behavior in GFAP-positive tanycytes. Genetic inactivation of Glut2 in GFAP-expressing tanycytes was performed using Cre/Lox technology. The efficiency of GFAP-tanycyte targeting was analyzed in the anteroposterior and dorsoventral axes by evaluating GFP fluorescence. Feeding behavior, hormonal levels, neuronal activity using c-Fos, and neuropeptide expression were also analyzed in the fasting-to-refeeding transition. In basal conditions, Glut2-inactivated mice had normal food intake and meal patterns. Implementation of a preceeding fasting period led to decreased total food intake and a delay in meal initiation during refeeding. Additionally, Glut2 inactivation increased the number of c-Fos-positive cells in the ventromedial nucleus in response to fasting and a deregulation of Pomc expression in the fasting-to-refeeding transition. Thus, a GLUT2-dependent glucose-sensing mechanism in GFAP-tanycytes is required to control food consumption and promote meal initiation after a fasting period.

Protein multi-scale organization through graph partitioning and robustness analysis: application to the myosin-myosin light chain interaction.

Despite the recognized importance of the multi-scale spatio-temporal organization of proteins, most computational tools can only access a limited spectrum of time and spatial scales, thereby ignoring the effects on protein behavior of the intricate coupling between the different scales. Starting from a physico-chemical atomistic network of interactions that encodes the structure of the protein, we introduce a methodology based on multi-scale graph partitioning that can uncover partitions and levels of organization of proteins that span the whole range of scales, revealing biological features occurring at different levels of organization and tracking their effect across scales. Additionally, we introduce a measure of robustness to quantify the relevance of the partitions through the generation of biochemically-motivated surrogate random graph models. We apply the method to four distinct conformations of myosin tail interacting protein, a protein from the molecular motor of the malaria parasite, and study properties that have been experimentally addressed such as the closing mechanism, the presence of conserved clusters, and the identification through computational mutational analysis of key residues for binding.

Body composition after endogenous (Cushing's syndrome) and exogenous (rheumatoid arthritis) exposure to glucocorticoids.

Exposure to chronic glucocorticoid (GC) excess determines changes in body composition. The aim of the study was to compare body composition in women exposed to endogenous hypercortisolism (Cushing's syndrome, CS), exogenous glucocorticoid treatment (rheumatoid arthritis, RA) and controls. Fifty-one CS women, 26 RA women treated with low-dose prednisone (5 mg/day or 10 mg/2 days), and 78 female controls were included. Fourteen CS patients were hypercortisolemic, 37 in remission (10 required hydrocortisone substitution after surgery). Body composition parameters were measured by dual-energy X-ray absorptiometry scanning (DEXA). RA patients had a greater waist-hip ratio (WHR) (p<0.01), less lean body mass (LBM) (p<0.01), and lumbar bone mineral density (BMD) (p<0.01) than controls. CS patients, globally and those with cured disease, had more total fat (both percentage and kg) and trunk fat percentage, and less whole body-BMD than RA patients (p<0.05, p<0.01, p<0.05, respectively). Active CS patients had less whole body-BMD and more LBM than RA patients (p<0.05, p=0.01, respectively). Cured CS patients not taking hydrocortisone had more total fat [both percentage (p<0.05) and kg (p<0.05)], trunk fat percentage (p<0.05), lumbar BMD (p<0.01) than RA patients. Cured CS patients requiring hydrocortisone only differed from RA patients by smaller WHR (p<0.01). All the differences in BMD disappeared when the data were reanalyzed including only the estrogen-deficient groups. Hypercortisoliof CS determines an irreversible increase in body fat, greater than in RA. Endogenous and exogenous exposure to GC negatively affects body composition by increasing the WHR. There appears to be no additional effect on BMD in estrogen-deficient women.

Diagnosis of cure in Cushing's syndrome: lessons from long-term follow-up.

It is generally assumed that endocrine 'cure' of hypercortisolism after successful treatment for Cushing's syndrome (CS) is associated with reversal of increased morbidity and mortality, typical of the active disease. However, recent data do not support this idea; increased cardiovascular risk is still present 5 years after endocrine cure, and health-related quality of life (HRQoL), although improved when compared to the active phase of hypercortisolism, is still impaired when compared to normal population. Abnormal body composition typical of hypercortisolism (i.e., increased total and trunk fat, reduced bone mass and lean body mass) is not completely normalized, even years after controlling hypercortisolism. Thus, control of hypercortisolism in CS does not normalize HRQoL, long-term cardiovascular risk and morbidity, body composition nor some metabolic parameters. Whether the same occurs in patients exposed to pharmacological doses of exogenous glucocorticoids, and whether the body composition abnormalities associated with the exposure to exogenous glucocorticoids are reversible or not, are worth considering.

Anatomy of the brainstem: a gaze into the stem of life.

The brainstem has an ectodermal origin and is composed of 4 parts: the diencephalon, mesencephalon, pons, and medulla oblongata. It serves as the connection between the cerebral hemispheres with the medulla and the cerebellum and is responsible for basic vital functions, such as breathing, heartbeat blood pressure, control of consciousness, and sleep. The brainstem contains both white and gray matter. The gray matter of the brainstem (neuronal cell bodies) is found in clumps and clusters throughout the brainstem to form the cranial nerve nuclei, the reticular formation, and pontine nuclei. The white matter consists of fiber tracts (axons of neuronal cells) passing down from the cerebral cortex--important for voluntary motor function--and up from peripheral nerves and the spinal cord--where somatosensory pathways travel--to the highest parts of the brain. The internal structure of brainstem, although complex, presents a systematical arrangement and is organized in 3 laminae (tectum, tegmentum, and basis), which extend its entire length. The motor pathway runs down through the basis, which is located at the most anterior part. The cranial nerve nuclei are settled into the middle layer (the tegmentum), just in front of the 4th ventricle and are placed, from medial to lateral, on the basis of their function: somatic motor, visceral motor, visceral sensory, and somatic sensory. All the somatosensory tracts run upward to the thalamus crossing the tegmentum in front of the cranial nerve nuclei. The tectum, formed by the quadrigeminal plate and the medullary velum, contains no cranial nuclei, no tracts and no reticular formation. The knowledge of precise anatomical localization of a lesion affecting the brainstem is crucial in neurological diagnosis and, on this basis, is essential to be familiar with the location of the mayor tracts and nuclei appropriately. Nowadays, current magnetic resonance imaging techniques, although still macroscopic, allow the fine internal structure of the brainstem to be viewed directly and make it possible to locate the main intrinsic structures that justify the symptoms of the patient. In this article we discuss the anatomy of the brainstem and highlight the features and landmarks that are important in interpreting magnetic resonance imaging.

Thyrotoxic periodic paralysis: a case report and literature review.

We describe a 37-year-old man with a 4-month history of episodic muscular weakness, involving mainly lower-limbs. Hypokalemia was documented in one episode and managed with intravenous potassium chloride. Hyperthyroidism was diagnosed 4 months after onset of attacks because of mild symptoms. The patient was subsequently diagnosed as having thyrotoxic periodic paralysis associated with Graves' disease. Treatment with propranolol and methimazol was initiated and one year later he remains euthyroid and symptom free. Thyrotoxic periodic paralysis is a rare disorder, especially among Caucasians, but it should always be considered in patients with acute paralysis and hypokalemia, and thyroid function should be evaluated.

Toxicity and characterization of cholinesterase-inhibition induced by diisopropyl fluorophosphate in Artemia salina larvae.

The acute toxicity of diisopropyl fluorophosphate (DFP) on three age classes of Artemia salina was evaluated. An increase in toxicity of this organophosphorous (OP) compound was found following longer development of A. salina larvae. The effects of pretreatment with the non-selective muscarinic antagonist atropine, the two reversible acetylcholinesterase inhibitors physostigmine and pyridostigmine, and the cholinesterase-reactivating oxime 2-pyridine aldoxime methoiodide (2-PAM), as individual and combined pretreatments, on DFP-induced lethality in 24h Artemia were also investigated. The lethal action of DFP was not prevented by pretreatment of 24h Artemia with atropine, physostigmine, and pyridostigmine, while 2-PAM proved effective against intoxication with this OP compound. The inhibitory effects of combinations of atropine (10(-5)M) plus 2-PAM or physostigmine were greater than those elicited by either drug alone, with the maximum protection afforded being 100%. Pretreatment with 2-PAM (10(-6)M) plus physostigmine or pyridostigmine was ineffective. These results suggest that the combinations of atropine plus 2-PAM or physostigmine are effective in the prevention of the lethal effects induced by DFP in A. salina larvae.

[Role of health-related quality of life in clinical practice: subjectivity in evidence]. / Calidad de vida relacionada con la salud, su lugar en la práctica clínica: lo subjetivo en la evidencia.

The incorporation of subjective elements provided by the patient to medical practice constitutes a new bridge between two historical movements of human culture: naturalism and rationalism. In recent years, medical sciences have identified these movements as patient-centered medicine and evidence-based medicine. By revisiting the paradigms of medicine, theories of causality, doctor-patient relationship models, bioethical principles and examples of clinical studies, their role and meaning has been revised. The applicability for neuroendocrinology has been demonstrated by validating therapeutic results with measurement of health-related quality of life. The need to incorporate the patient's subjective perception poses a challenge that, when accepted, can advance the understanding and care of human beings--ultimately rational, thinking subjects, and therefore subjective--as our species has been defined.

Role of neuronal voltage-gated K(+) channels in the modulation of the nitrergic neurotransmission of the pig urinary bladder neck.

BACKGROUND AND PURPOSE: As nitric oxide (NO) plays an essential role in the inhibitory neurotransmission of the bladder neck of several species, the current study investigates the mechanisms underlying the NO-induced relaxations in the pig urinary bladder neck. EXPERIMENTAL APPROACH: Urothelium-denuded bladder neck strips were dissected and mounted in isolated organ baths containing a physiological saline solution at 37 degrees C and continuously gassed with 5% CO(2) and 95% O(2), for isometric force recording. The relaxations to transmural nerve stimulation (EFS), or to exogenously applied acidified NaNO(2) solution were carried out on strips pre-contracted with phenylephrine, and treated with guanethidine and atropine, to block noradrenergic neurotransmission and muscarinic receptors, respectively. KEY RESULTS: EFS (0.2-1 Hz) and addition of acidified NaNO(2) solution (1 microM-1 mM) evoked frequency- and concentration-dependent relaxations, respectively. These responses were potently reduced by the blockade of guanylate cyclase and were not modified by the K(+) channel blockers iberiotoxin, charybdotoxin, apamin or glibenclamide. The voltage-gated K(+) (Kv) channels inhibitor 4-aminopyridine, greatly enhanced the nitrergic relaxations evoked by EFS, but did not affect the NaNO(2) solution-induced relaxations. CONCLUSIONS AND IMPLICATIONS: NO, whose release is modulated by pre-junctional Kv channels, relaxes the pig urinary bladder neck through a mechanism dependent on the activation of guanylate cyclase, in which post-junctional K(+) channels do not seem to be involved. Modulation of Kv channels could be useful in the therapy of the urinary incontinence produced by intrinsic sphincteric deficiency.

Prótesis unicompartimental de rodilla / Unicompartmental knee prosthesis

Resumen: En un paciente con artrosis unicompartimental de rodilla grave, donde se agotaron los tratamientos conservadores que tiene clínica dolorosa localizada en el lado afectado y con alteración del eje reductible, la prótesis unicompartimenal de rodilla (PUR) es la primera opción para nuestro grupo de trabajo. Dentro del estudio para confirmar el diagnóstico y planificar la cirugía destacan las radiografías de rodillas con carga, radiografía de Rosenberg y telerradiografías de extremidades inferiores. El objetivo de la cirugía es reemplazar la zona afectada, restituyendo la anatomía con un adecuado balance de partes blandas. Respecto al alineamiento el desafío es no sobrecargar el lado contrario ni tampoco el de la prótesis. Existen de platillo móvil y fijo y aunque los resultados clínicos y de supervivencia son semejantes, en los últimos años con la incorporación de la cirugía robótica, la balanza se ha inclinado para el uso de los platillos fijos. En pacientes con artrosis unicompartimental los resultados clínicos y funcionales son mejores con PUR y tiene menos complicaciones que cuando se usan prótesis totales (PTR). La supervivencia estudiada en registros es menor que para PTR, pero cuando se usa en centros de alto flujo en que el porcentaje de PUR es cercano a un tercio del total con una estricta selección de pacientes, la duración es tan buena como en la PTR.

Abstract: In a patient with severe unicompartmental knee osteoarthritis where conservative treatments have been exhausted, with painful symptoms located on the affected side and with a reducible axis, the unicompartmental knee prosthesis (UKP) is the first option for our work group. Within the study to confirm the diagnosis and plan the surgery, weight-bearing knee x-rays, Rosenberg x-rays, and teleradiographs of the lower extremities stand out. The objective of surgery is to replace the affected area, restoring the anatomy with an adequate balance of soft tissues. Regarding alignment, the challenge is not to overload the opposite side or that of the prosthesis. There are mobile and fixed plates and although the clinical and survival results are similar, in recent years with the incorporation of robotic surgery, the balance has tipped towards the use of fixed plates. The clinical and functional results are better and there are fewer complications than when total knee prostheses (TKP) are used in the same type of patients. The survival studied in registries is lower than for TKP, but when used in high-flow centers where the percentage of UKP is close to a third of the total with strict patient selection, the duration is as good as in PTR.

[Caring for perilesional skin or skin having a lesion risk]. / Cuidados en piel perilesional o con riesgo de lesión.

In order to increase the clinical and scientific evidence of the Hyperoxygenated Fatty Acids (HFA) in emulsion preparation for skin care, this study considers to evaluate prospectively how it influences in the state of the periwound skin (when there are active lesions) or in which it presents a high risk of lesion production.

[Effect of sports training in angular compartment of the lower limbs in children footballers aged 11 to 12 years old]. / Efecto de un entrenamiento deportivo en el compartimiento angular de la extremidad inferior en niños futbolistas de entre 11 y 12 años.

Anterior Cruciate Ligament (ACL) injury is an important cause of days lost in athletes. Most ACL injuries are non-contact and are associated with biomechanical risk factors that increase tension in the ACL: increased knee valgus (KV) and hip flexion (HF) and decreased flexion of knee (KF). Muscle around the knee contributes to knee stability, so fatigue produced by exercise could alter knee balance, increasing LCA tension. The aim of the study is to determine the angular behavior before and after a physical load for CF, RR and RV in children born in 2002-2003. A non-randomized clinical trial was conducted. The sample consisted of 50 students from soccer schools born between 2002 and 2003. The angular behavior of CF, RR and VR was compared, before and after performing standardized training. The angular behavior was measured by performing the DJ test with data obtained by inertial sensors. After exercise, the 3 variables increased, but only HF reached significant difference. Other important finding was the difference found in KV between the dominant leg and the support limb, at both times: rest and post exercise. It was concluded that the angular behavior of CF increases significantly in both limbs post-exercise and that preventive measures should be applied for the management of valgus in the supporting limb.

La lesión del ligamento cruzado anterior (LCA) constituye una causa importante de reposo en deportistas. En su mayoría, las lesiones del LCA no requieren contacto externo y se asocian con factores de riesgo biomecánicos que aumentan la tensión en el LCA: el aumento del ángulo de valgo de rodilla (VR), flexión de cadera (FC) y una menor flexión de rodilla (FR). El LCA requiere cooperación de los grupos musculares perirrodilla, por lo que la fatiga producida por el ejercicio alteraría el balance y pondría en riesgo a este ligamento. El objetivo del estudio es determinar el comportamiento angular antes y después de una carga física para FC, FR y VR en niños nacidos en los años 2002-2003. Se realizó un estudio clínico no aleatorizado. La muestra consistió en 50 alumnos de escuelas de fútbol nacidos en los años 2002 y 2003. Se comparó el comportamiento angular de FC, FR y VR antes y después de realizar un entrenamiento estandarizado. El comportamiento angular fue medido a través de la realización de la prueba DJ, con datos obtenidos por sensores inerciales. Posterior al ejercicio, aumentaron las tres variables; sólo fue significativo el incremento en la flexión de cadera. Por otra parte, destaca la diferencia encontrada tanto en reposo como postejercicios entre la extremidad de apoyo y la hábil en el peak de valgo angular. Se concluyó que el comportamiento angular de FC aumenta significativamente en ambas extremidades postentrenamiento y que se deben aplicar medidas preventivas para el manejo del valgo en la extremidad de apoyo.

Neuronal and smooth muscle receptors involved in the PACAP- and VIP-induced relaxations of the pig urinary bladder neck.

BACKGROUND AND PURPOSE: As pituitary adenylate cyclase-activating polypeptide 38 (PACAP 38)- and vasoactive intestinal peptide (VIP) are widely distributed in the urinary tract, the current study investigated the receptors and mechanisms involved in relaxations induced by these peptides in the pig bladder neck. EXPERIMENTAL APPROACH: Urothelium-denuded strips were suspended in organ baths for isometric force recordings and the relaxations to VIP and PACAP analogues were investigated. KEY RESULTS: VIP, PACAP 38, PACAP 27 and [Ala(11,22,28)]-VIP produced similar relaxations. Inhibition of neuronal voltage-gated Ca(2+) channels reduced relaxations to PACAP 38 and increased those induced by VIP. Blockade of capsaicin-sensitive primary afferents (CSPA), nitric oxide (NO)-synthase or guanylate cyclase reduced the PACAP 38 relaxations but failed to modify the VIP responses. Inhibition of VIP/PACAP receptors and of voltage-gated K(+) channels reduced PACAP 38 and VIP relaxations, which were not modified by the K(+) channel blockers iberiotoxin, charybdotoxin, apamin or glibenclamide. The phosphodiesterase 4 inhibitor rolipram and the adenylate cyclase activator forskolin produced potent relaxations. Blockade of protein kinase A (PKA) reduced PACAP 38- and VIP-induced relaxations. CONCLUSIONS AND IMPLICATIONS: PACAP 38 and VIP relax the pig urinary bladder neck through muscle VPAC(2) receptors linked to the cAMP-PKA pathway and involve activation of voltage-gated K(+) channels. Facilitatory PAC(1) receptors located at CSPA and coupled to NO release, and inhibitory VPAC receptors at motor endings are also involved in the relaxations to PACAP 38 and VIP, respectively. VIP/PACAP receptor antagonists could be useful in the therapy of urinary incontinence produced by intrinsic sphincter deficiency.

Prediction of allosteric sites and mediating interactions through bond-to-bond propensities.

Allostery is a fundamental mechanism of biological regulation, in which binding of a molecule at a distant location affects the active site of a protein. Allosteric sites provide targets to fine-tune protein activity, yet we lack computational methodologies to predict them. Here we present an efficient graph-theoretical framework to reveal allosteric interactions (atoms and communication pathways strongly coupled to the active site) without a priori information of their location. Using an atomistic graph with energy-weighted covalent and weak bonds, we define a bond-to-bond propensity quantifying the non-local effect of instantaneous bond fluctuations propagating through the protein. Significant interactions are then identified using quantile regression. We exemplify our method with three biologically important proteins: caspase-1, CheY, and h-Ras, correctly predicting key allosteric interactions, whose significance is additionally confirmed against a reference set of 100 proteins. The almost-linear scaling of our method renders it suitable for high-throughput searches for candidate allosteric sites.