Fungal community and physicochemical profiles of ripened cheeses from the Canastra of Minas Gerais, Brazil.

Canastra's Minas artisanal cheese [QMA (Minas artisanal cheese)] is a protected geographical indication traditional food. The influence of fungi on the cheese ripening process is of great importance. This study aimed to apply culture-dependent and -independent methods to determine the mycobiota of QMA produced in the Canastra region, as well as to determine its physicochemical characteristics. Illumina-based amplicon sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were the culture-independent methods used. The physicochemical analysis results showed that the QMA has a moisture content ranging 18.4-28.2%, fat content ranging 20.5-40%, sodium chloride percentage of approximately 0.9%, and pH ranging 5.2-5.5. The population of fungi ranged between 6.3 and 8 log colony-forming unit/g. Fusarium spp., Geotrichum candidum, Paecilomyces spp., Trichosporon coremiiforme, Candida catenulata, Aspergillus spp., Trichosporon japonicum, Aspergillus oryzae, Kluyveromyces spp., Torulaspora spp., and Debaryomyces spp. were the most prevalent fungi. The methods used to evaluate the mycobiota provide a better understanding of which species are present in the final product and eventually contribute to the characteristics of QMA. Geotrichum candidum and C. catenulata were identified as promising species for future studies on product quality.

Development of a Prototype E-Textile Sock.

The aim of this work is to design and develop a sensorized sock in Electronic Textile (ET), SWEET-Sock. The device has been realized by three textile sensor placed in a specific points of plantar arch and an accelerometer unit, both embedded and connected by conductive thread. The sensors allows the acquisition of plantar pressure and acceleration signals deriving from the motion of the lower limbs. The detected biosignals have been condictionated by a voltage divider and then were acquired through a LilyPad Arduino microcontroller and transmitted using the Simblee BLE technology to a custom made mobile app. Data were afterwards uploaded through a smartphone on a dropbox cloud where a custom made MATLAB GUI platform has been developed for further digital signal processing of main biomechanical parameters of clinical interest in postural and gait analysis.

Changes of excitatory and inhibitory neurotransmitters in the colon of rats underwent to the wrap partial restraint stress.

BACKGROUND: Animal models proposed to reproduce some of the human irritable bowel syndrome (IBS) symptoms are based on the hypothesis that psychosocial stressors play a pivotal role in the IBS etio-pathology. We investigated the wrap restraint stress (WRS) model with the aim to analyze the morphological changes of the entire colonic wall of these animals that showed some of the human IBS symptoms such as visceral hypersensitivity. METHODS: Male Wistar rats were used and WRS was maintained for 2 h. Abdominal contractions (AC) were recorded in the colon-rectum by balloon distension. Fecal pellets were quantitated. Colonic specimens were examined by routine histology, immunohistochemistry and western blot. KEY RESULTS: WRS animals were characterized by: (i) increase in AC number and fecal pellets mean weight; (ii) clusters of mononucleated cells, increase in eosinophilic granulocytes and mast cells in the mucosa; (iii) increase in CGRP-immunoreactive (IR) nerve fibers in the lamina propria; (iv) decrease in myenteric NK1r-IR and nNOS-IR neurons and in submucous nNOS-IR neurons; (v) decrease in SP-IR nerve fibers in the muscle wall; (vi) reduction in S100ß-IR glia in the entire colonic wall; (vii) increase in CRF1r-IR myenteric neurons; (viii) no change in ChAT-IR neurons, smooth muscle cells and interstitial cells of Cajal. CONCLUSIONS AND INFERENCES: The present results support the consistency of the WRS as a potential model where part of the human IBS signs and symptoms are reproduced. The changes in glial cells and in excitatory and inhibitory neurotransmitters might represent the substrate for the dysmotility and hypersensitivity.

Changes in nitrergic and tachykininergic pathways in rat proximal colon in response to chronic treatment with otilonium bromide.

BACKGROUND: Otilonium bromide (OB) is used as a spasmolytic drug in the treatment of the functional bowel disorder irritable bowel syndrome. Although its acute effects on colonic relaxation are well-characterized, little is known about the effects of chronic administration of OB on enteric neurons, neuromuscular transmission, and interstitial cells of Cajal (ICC), key regulators of the gut function. METHODS: Adult Sprague-Dawley rats were treated with OB in drinking water at a dose of 2 mg/kg for 30 days. The colons of OB-treated and age-matched control rats were studied by confocal immunohistochemistry to detect immunoreactivity (IR) in myenteric plexus neurons for nitrergic and tachykininergic markers, and also by microelectrode electrophysiology. KEY RESULTS: Using immunohistochemistry, chronic OB administration did not change total neuron number, assessed by anti-Hu IR, but resulted in a significant increase in NK1 receptor positive neurons, a decrease in neuronal nitric oxide synthase expressing neurons, and a reduction in volume of substance P in nerve fibers in the myenteric plexus. Chronic OB administration potentiated inhibitory and excitatory junction potentials evoked by repetitive electrical field stimulation. The various types of colonic ICC, detected by Kit IR, were not altered nor were slow waves or smooth muscle membrane potential. CONCLUSIONS & INFERENCES: Chronic treatment with OB caused significant changes in the nitrergic and tachykinergic components of the myenteric plexus and in both inhibitory and excitatory neurotransmission in the rat colon.

Chronic treatment with otilonium bromide induces changes in L-type Ca²âº channel, tachykinins, and nitric oxide synthase expression in rat colon muscle coat.

BACKGROUND: Otilonium bromide (OB) is a quaternary ammonium derivative used for the treatment of intestinal hypermotility and is endowed with neurokinin2 receptor (NK2r) antagonist and Ca²âº channel blocker properties. Therefore, the possibility that OB might play a role in the neurokinin receptor/Substance-P/nitric oxide (NKr/SP/NO) circuit was investigated after chronic exposition to the drug. METHODS: Rats were treated with OB 2-20 mg kg⁻¹ for 10 and 30 days. In the proximal colon, the expression and distribution of muscle NOsynthase 1 (NOS1), NK1r, NK2r, SP and Cav 1.2 subunit (for L-type Ca²âº channel) and the spontaneous activity and stimulated responses to NK1r and NK2r agonists were investigated. KEY RESULTS: Immunohistochemistry showed a redistribution of NK1r and L-type Ca²âº channel in muscle cells with no change of NK2r at 30 days, a significant increase in muscle NOS1 expression at 10 days and a significant decrease in the SP content early in the ganglia and later in the intramuscular nerve fibers. Functional studies showed no change in spontaneous activity but a significant increase in maximal contraction induced by NK1r agonist. CONCLUSIONS & INFERENCES: Chronic exposition to OB significantly affects the NKr/SP/NO circuit. The progressive decrease in SP-expression might be the consequence of the persistent presence of OB, the increase of NOS1 expression in muscle cells at 10 days in an attempt to guarantee an adequate NO production, and, at 30 days, the redistribution of the L-type Ca²âº channel and NK1r as a sign to compensate the drug channel block by re-cycling both of them. The physiological data suggest NK1r hypersensitivity.

Comparison between the effects of lisinopril and losartan on the cougn reflex in anesthetized and awake rabbits.

The aim of the present study was to analyze differences in cough induction between losartan and lisinopril in both anaesthetized and awake rabbits, i.e., under conditions in which the influences of higher brain areas on the cough reflex are strongly reduced or abolished. Losartan (500 µg/kg), lisinopril (100 µg/kg) and NaCl 0.9% saline solution (vehicle) were administered by intravenous injections. Animals were randomly assigned to the different experimental treatments. The cough reflex was induced by chemical (citric acid) and/or mechanical stimulation of the tracheobronchial tree. In anaesthetized rabbits, losartan and lisinopril caused similar hypotensive effects. Lisinopril, but not losartan, increased the cough response induced by both mechanical and chemical stimulation due to increases in the cough number, i.e. the number of coughs induced by each stimulation challenge. In awake animals, only lisinopril significantly increased the cough number. The results support the notion that cough potentiation induced by losartan, and possibly other sartans, is lower than that induced by most angiotensin-converting enzyme inhibitors despite the reduction or complete absence of higher brain functions. In this connection, the comparison between present results and our previous findings on ramipril and zofenopril shows that losartan and zofenopril display similar cough-inducing potency, much lower than that of lisinopril and ramipril.

Fitoterápicos na odontologia: estudo etnobotânico na cidade de Manaus / Phytotherapics in Odontology: ethnobotanical study in Manaus

O reconhecimento oficial da fitoterapia na odontologia no Brasil veio acompanhado de diversas lacunas na pesquisa científica e na utilização de plantas medicinais, especificamente para espécies vegetais com aplicação nas doenças da cavidade oral. O estado do Amazonas, especificamente a cidade de Manaus, não possui um diagnóstico da aplicabilidade de plantas medicinais nos serviços de atenção Odontológica. Esse estudo propôs realizar um estudo do tipo quali-quantitativo, descritivo e exploratório visando o levantamento da comercialização de plantas medicinais e o estudo etnobotânico para identificação das principais plantas medicinais indicadas e utilizadas nas patologias orais nos atendimentos odontológicos ambulatoriais na cidade de Manaus. Foram entrevistados 197 usuários do serviço odontológico, 150 Cirurgiões-Dentistas, e 47 comerciantes credenciados na prefeitura Municipal de Manaus. A amostragem foi realizada por acessibilidade ou conveniência para os Cirurgiões-Dentistas, enquanto os usuários foram selecionados através de amostragem aleatória simples, e os comerciantes de plantas medicinais foram entrevistados em sua totalidade. Os resultados demonstraram a existência de comercialização de plantas medicinais para patologias orais, destacando-se as seguintes espécies: Pedra ume cãa (Aulomyrcia sphareocarpa), Crajiru (Arrabidae chica), além da planta, sem identificação botânica, conhecida popularmente como Sara tudo. Entre os Cirurgiões-Dentistas e entre os pacientes, apenas 8% e 7,61%, respectivamente, utilizaram plantas medicinais para alterações patológicas orais. Os autores concluíram que as plantas medicinais comercializadas na cidade de Manaus são utilizadas de maneira empírica e que, apesar da Política Nacional de Práticas Integrativas e Complementares (PNPIC), novas políticas públicas de saúde devem inserir plantas medicinais e fitoterápicos de uso oral na rede pública de saúde na cidade de Manaus.

The official recognition of herbal medicine in dentistry in Brazil was accompanied by several gaps in scientific research and the use of medicinal plants, specifically for plant species applied to diseases of the oral cavity. The state of Amazonas, specifically the city of Manaus, does not have a diagnosis of applicability of medicinal plants in Dental care services. This research intended to conduct a qualitative and quantitative-type, descriptive and exploratory study in order to conduct a survey on the commercialization of medicinal plants and an ethnobotanical study to identify the main medicinal plants indicated and used in oral pathologies in outpatient dental care in the city of Manaus. We interviewed 197 users of dental services, 150 dentists, and 47 accredited traders in the Municipality of Manaus. The sampling was performed by accessibility or convenience for surgeon-dentists, while users were selected through simple random sampling, and all traders of medicinal plants were interviewed. The results demonstrated that medicinal plants for oral diseases are commercialized, among which the following species are highlighted: Pedra ume Caa (Aulomyrcia sphareocarpa), Crajiru (Arrabidaea chica), in addition to an unidentified botanical plant popularly known as Sara Tudo. 8% of dentists and 7.61% of patients used medicinal plants for oral pathological changes. The authors concluded that medicinal plants commercialized in the city of Manaus are used empirically, and that, despite the rules from the National Policy on Integrative and Complementary Practices (PNPIC), new public health policies must add medicinal plants and herbal medicines for oral use to the public health system of the city of Manaus.

Effect of otilonium bromide and ibodutant on the internalization of the NK2 receptor in human colon.

BACKGROUND: The present aim was to study the modulation of NK2 receptor internalization by two compounds, the spasmolytic otilonium bromide (OB) endowed with NK2 receptor antagonistic properties and the selective NK2 receptor antagonist ibodutant. METHODS: Full-thickness human colonic segments were incubated in the presence of OB (0.1-10 µmol L(-1)) or ibodutant (0.001-0.1 µmol L(-1)), with or without the NK2 receptor selective agonist [ßAla8]NKA(4-10) and then fixed in 4% paraformaldehyde. Cryosections were processed for NK2 receptor immunohistochemical revelation. Quantitative analysis evaluated the number of the smooth muscle cells that had internalized the NK2 receptor. KEY RESULTS: Immunohistochemistry revealed that in basal condition, the NK2 receptor was internalized in about 23% of total smooth muscle cells. The exposure to the selective NK2 receptor agonist induced internalization of the receptor in more than 77% of the cells. Previous exposure to both OB or ibodutant, either alone or in the presence of the agonist, concentration-dependently reduced the number of the cells with the internalized receptor. CONCLUSIONS & INFERENCES: Both OB and ibodutant antagonize the internalization of the NK2 receptor in the human colon. As NK2 receptors are the predominant receptor mediating spasmogenic activity of tachykinins on enteric smooth muscle, we hypothesize that the antagonistic activity found for both OB and ibodutant should play a specific therapeutic role in gut diseases characterized by hypermotility.

The effect of structure and a secondary carbon source on the microbial degradation of chlorophenoxy acids.

Pseudomonas putida, Aspergillus niger, Bacillus subtilis, Pseudomonas fluorescens, Sphingomonas herbicidovorans and Rhodococcus rhodochrous growing on glucose in a medium containing one of three chlorophenoxy acids at a concentration of 0.1 g L(-1) (clofibric acid, (R)-2-(4-chloro-2-methylphenoxy)propionic acid (mecoprop or MCPP) and 4-chloro-2-methylphenoxyacetic acid (MCPA)) degraded these compounds to varying degrees; from nonmeasurable to almost complete removal. These results with the addition of glucose (2.5 g L(-1)) as an easy to use carbon source indicated the formation of metabolites different from results reported in the literature for growth studies in which the chlorophenoxy acid was the sole carbon source. The metabolite, 4-chloro-2-methylphenol, which had been reported previously, was only observed in trace amounts for MCPP and MCPA in the presence of S. herbicidovorans and glucose. In addition, three other compounds (M1, M3 and M4) were observed. It is suggested that these unidentified metabolites resulted from ring opening of the metabolite 4-chloro-2-methylphenol (M2). The rate of biodegradation of the chlorophenoxy acids was influenced by the degree of steric hindrance adjacent to the internal oxygen bond common to all three compounds. The most hindered compound, clofibric acid, was converted to ethyl clofibrate by R. rhodochrous but was not degraded by any microorganisms studied. The more accessible internal oxygen bonds of the other two chlorophenoxy acids, MCPP and MCPA, were readily broken by S. herbicidovorans.

Influence of tachykinin NK2 receptors on intestinal sensitivity and motility in newborn rats.

The effect of tachykinin neurokinin NK(2) receptors activation on intestinal propulsion and colorectal sensitivity was studied in 7-15 days old newborn rats. In a first set of experiments investigating the intestinal transit, the selective NK(2) receptor agonist, [betaAla(8)]NKA-(4-10) was used. It produced an increase of the small intestinal transit measured by charcoal test of 54%, that was inhibited in a dose-dependent manner by nepadutant ([N(4)-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-L-asparaginyl-L-aspartyl-L-tryptophyl-L-phenylalanyl-L-2,3-diaminopropionyl-L-leucyl]-C-4.2-N-3.5-lactam-C-1.6-N-2.1-lactam), a known selective NK(2) receptor antagonist, orally administered 2-48 h before the challenge with the NK(2) receptor agonist. Nepadutant did not affect the basal intestinal propulsion and showed a good oral bioavailability and long duration of action. In another set of experiments investigating visceral sensitivity, a fixed distension volume of a balloon inserted intrarectally in 14-15 days old newborns rats produced abdominal contractions (AC) that were increased after colonic application of acetic acid (50 microl, 0.5%). In this latter condition nepadutant, at 0.5 and 2.5 mg/kg p.o., significantly reduced the resulting AC. In control rats, untreated with acetic acid, nepadutant did not affect AC evoked by colorectal distension. These findings show for the first time two models to assess intestinal motility and visceral sensitivity in newborn rats and indicate nepadutant as a valuable tool to assess the role of NK(2) receptors in the intestinal propulsive and nociceptive activity in infants.

Role of calcitonin gene-related Peptide in gastric mucosal defence and healing.

Calcitonin gene-related peptide (CGRP) is a polypeptide produced by alternative processing of calcitonin gene transcripts and is endowed with important systemic physiological effects. The recent characterization of its receptor and the discovery of stable antagonists has addressed them in the indication migraine. Beside this, several studies have been focused on role of CGRP at gastric level. CGRP is considered a marker of afferent fibers in the upper gastrointestinal tract being almost completely depleted following treatment with the selective neurotoxin capsaicin that targets these fibers via transient receptor potential vanilloid of type-1. The exogenous administration of the peptide was able to afford protection against experimental ulcers induced by an increase in gastric secretion. The use of CGRP knockout mice has let to characterize the endogenous role of CGRP showing that the local release of this neuropeptide protects from ethanol injury and favours ulcer healing. Decreased levels of gastric CGRP-like immunoreactivity (li) were observed during acetic acid-, cysteamine-, concentrated ethanol- or water immersion stress-ulcers. Restoration of CGRP-li was found in animals bearing ulcers in healing status and delayed healing in mice knockout to CGRP. CGRP was able to release somatostatin from gastric D cells but its main effects on the stomach homeostasis rely on local vasodilator action during increased acid-back diffusion.

Spatial and temporal expression of tachykinins and NK1- and NK2-receptor gene during TNB induced colitis in rats.

The distribution and modification of the tachykinins Substance P (SP) and neurokinin A (NKA), their precursor beta-preprotachykinins (beta-PPT) and the receptors involved in their activity, NK-1 and NK-2, were studied in trinitrobenzensulphonic acid (TNB) colitis. Rats were intrarectally treated with a 120 mg/ml of TNB solution and sacrificed at various times after colitis induction. During the acute phases of colitis, a marked decrease in tissue SP and NKA levels were observed along with an increased transcription of beta-PPT mRNA in the neurons of the myenteric plexus and an increased myeloperoxidase activity, which is an index of the tissue's inflammatory status. De novo expression of both NK(1) and NK(2) receptor mRNA was observed during the acute phase of TNB-colitis in mesenchymal cells around dilated submucosal vessels but their expression in smooth muscle cells of the muscularis mucosae and propria was moderately down-regulated. The peptide levels, myeloperoxidase activity and gene expression of tachykinin receptors were then restored during the late phases (2-4 weeks after the apten administration) while beta-PPT mRNA remained highly expressed in the myenteric plexus ganglia showing that SP and NKA are involved in repair processes. These results point to the enhanced release of tachykinins during the initial phase of colitis and a restoration of this neuropeptide pool in the healing of the tissue.

FIGO Stage I endometrial carcinoma: evaluation of lung metastases and follow-up.

PURPOSE: The aim of our study was to evaluate the incidence of lung metastases in the follow-up of women submitted to surgery for endometrial carcinoma, in particular for FIGO Stage I which is the lowest risk stage for this metastatic site. METHODS: The study was conducted on 210 patients affected by FIGO Stage I endometrial cancer in the years 1990 to 2005 distributed as follows: 35 patients with Stage IA (limited to the endometrium), 150 patients with Stage IB (invasion up to and including half the myometrial thickness), 25 patients with Stage IC (invasion greater than half the myometrial thickness). They underwent follow-up. RESULTS: Only one patient out of the group studied has developed lung metastasis six years after surgery. She was staged as FIGO IB (T1b Mx G1). CONCLUSION: We are still following the cases and evaluating the biological behavior of this specific endometrial carcinoma and its reaction to further therapies. We are also looking for possible clinical characteristics in disagreement with those reported in the literature, which would thus make it necessary to reconsider the prognosis and therapy of this stage of disease.

A importância da notificação de acidentes por animais peçonhentos em Campo Grande - MS / The importance of the notification of accidents involving venomous animals in Campo Grande - MS

Este estudo de caráter quantitativo e descritivo objetivou demonstrar a importância da notificação de acidentes por animais peçonhentos no Hospital Regional de Mato Grosso do Sul em Campo Grande, comparando suas notificações no Centro Integrado de Vigilância Toxicológica (CIVITOX) e o Sistema de Informação de Agravos de Notificação (SINAN), a partir dos laudos médicos para emissão das autorizações de internações hospitalares, e investigar o conhecimento dos profissionais de saúde a respeito da rotina de notificação. Concluiu-se a existência de subnotificação, divergências nos registros dos sistemas de informação e o desconhecimento dos profissionais em notificá-los.

Biochemical basis of ischemic heart injury and of cardioprotective interventions.

Cardioprotective interventions are defined as interventions able to increase myocardial resistance to ischemia. The authors approach the issue of cardioprotection on the basis of the present knowledge about the biochemical mechanisms responsible for the injury produced by myocardial ischemia or ischemia-reperfusion. Reversible and irreversible injury are distinguished. The former is largely accounted for by the direct consequences of reduced ATP synthesis, which causes decreased ATP phosphorylation potential, acidosis and phosphate accumulation. The biochemical mechanisms leading to irreversible injury include osmotic overload, production of toxic lipid metabolites, cytosolic calcium overload, and generation of reactive oxygen species, which lead to membrane disruption, mitochondrial dysfunction and possibly to the activation of apoptotic pathways. The major effect of the classical cardioprotective agents (nitrates, beta adrenergic antagonists, calcium channel blockers) consists in affecting ATP demand/supply ratio in such a way as to delay the decrease in ATP phosphorylation potential. Other drugs have been introduced, which allegedly interfere directly with the mechanisms responsible for irreversible ischemic injury. These include 3-ketoacyl-CoA tiolase inhibitors, modulators of intracellular calcium channels, ionic exchanger inhibitors, free radical scavengers, caspase inhibitors, purinergic agonists, K(+)(ATP) channel openers, and modulators of mitochondrial permeability transition. The results obtained with these substances in experimental models and in the clinical setting are discussed. Special attention is devoted to angiotensin converting enzyme inhibitors, whose direct cardioprotective properties has recently been demonstrated.

Role of sensory neurons in restitution and healing of gastric ulcers.

It has been shown that capsaicin-sensitive afferent fibers play a crucial role in acute gastroprotection. Release of neurotransmitters such as calcitonin gene-related peptide (CGRP) and the consequent increase in mucosal blood flow have been identified as key factors in the protective effect of the stimulation of these fibers by capsaicin. Conversely the involvement of sensory nerves in the process of tissue repair after acute and chronic gastric mucosal damage has remained largely unexplored. Some studies, however, while demonstrating that the process of rapid repair (restitution) of the gastric mucosa damaged by ethanol is unaffected by capsaicin pretreatment, have shown that the recovery of gastric integrity after mucosal damage induced by sodium taurocholate or monochloramine, a known cytotoxic agent present in H. pylori patients, requires an intact sensory function and the maintenance of an adequate blood supply. In addition, a delayed healing (up to 1 week) of HCl-induced gastric lesions has been reported in capsaicin-deafferented rats, in association with a selective impairment of the hyperemic response to acid. Healing of gastric lesions induced by indomethacin, ischaemia and reperfusion, water restraint stress or concentrated ethanol was delayed in animals with functional ablation of sensory nerves. In a well-validated model, such as chronic gastric ulcers induced in rats by subserosal injection of acetic acid whose lesions last up to 4 weeks, the chemical ablation of sensory neurons negatively interferes with the process of chronic ulcer healing. The delay in ulcer healing was found to be associated with a persistent decrease in tissue levels of gastric CGRP and with a change of inflammatory mediators and growth factors, while gastric secretion and emptying were not concomitantly affected. Taken together, these data suggest that capsaicin-sensitive afferent nerves may play a role in the process of ulcer healing by mediating the hyperemic response through the release of CGRP and facilitating the acid disposal in the mucosa. From a therapeutic perspective, it is obvious that the compound acting on this system could have a role in the healing processes of the stomach damage.

Cervical ectopic pregnancy: clinical review.

Cervical pregnancy is a rare kind of intrauterine ectopic pregnancy. Diagnosis and treatment of cervical pregnancy have enormously changed in the last 15 years. Before 1980, diagnosis was made when dilation and curettage for presumed incomplete abortion resulted in sudden and uncontrollable hemorrhage. Hysterectomy was practiced in order to save the patient's life. Today, cervical pregnancy is diagnosed by ultrasound (US) during the Ist trimester of pregnancy, so that the patient's fertility can be preserved. Therefore any physician should consider the possibility of a cervical pregnancy in a woman with abdominal pain and vaginal bleeding during the first trimester of pregnancy. In this study we reviewed the literature on the epidemiology, etiology, diagnosis and treatments of cervical pregnancy.

Antioxidant and cardioprotective properties of the sulphydryl angiotensin-converting enzyme inhibitor zofenopril.

Zofenopril, a new potent sulphydryl angiotensin-converting enzyme (ACE) inhibitor, is characterized by high lipophilicity, selective cardiac ACE inhibition, and antioxidant and tissue protective activities. In vitro and in vivo experiments suggest that zofenopril exerts antioxidant properties at clinically achievable tissue concentrations. In endothelial cells, zofenopril enhances nitric oxide production, attenuates atherosclerotic lesion development and inhibits adhesion molecule expression by reducing reactive oxygen species. These peculiar characteristics are reflected in the drug's cardioprotective activity, which has been shown to be greater than that of non-sulphydryl ACE inhibitors. Cardiac hypertrophy was also reduced by chronic zofenopril administration, independently of its blood pressure-reducing effect. ACE inhibitors with a sulphydryl group could have an advantage in improving vascular function and reducing cardiac impairment compared with non-sulphydryl-containing ACE inhibitors. This could explain zofenopril's remarkable clinical efficacy post-infarction, and potentially beneficial use in prevention and therapy of cardiovascular diseases, such as atherosclerosis, thrombosis and heart failure.

Quaternary ammonium derivatives as spasmolytics for irritable bowel syndrome.

Quaternary ammonium derivatives such as cimetropium, n-butyl scopolammonium, otilonium and pinaverium bromide have been discovered and developed as potent spasmolytics of the gastrointestinal tract. Their pharmacological activity has been proven in both "in vivo" and "in vitro" studies of hypermotility. "In vitro" experiments showed that they possess antimuscarinic activity at nM level but only pinaverium and otilonium are endowed with calcium channel blocker properties. These latter compounds relaxed the gastrointestinal smooth muscle mainly through a specific inhibition of calcium ion influx through L-type voltage operated calcium channels. Molecular pharmacology trials have indicated that pinaverium and otilonium can bind specific subunits of the calcium channel in the external surface of the plasma membrane and in this way they block the machinery of the contraction. Recent evidence showed that otilonium is able to bind tachykinin NK(2) receptors and not only inhibits one of the major contractile agents but can reduce the activation of afferent nerves devoted to the passage of sensory signals from the periphery to the central nervous system. Thanks to their typical physico-chemical characteristics, they are poorly absorbed by the systemic circulation and generally remain in the gastrointestinal tract where they exert the muscle relaxant activity by a local activity. Some differences exists in the absorption among these compounds: both n-butyl scopolammonium and cimetropium are partially taken up in the bloodstream, pinaverium has a low absorption (8-10 %) but is endowed with an excellent hepato-biliary excretion and otilonium, which has the lowest absorption (3 %), is almost totally excreted by faeces. Quaternary ammonium derivatives are widely used for the treatment of irritable bowel syndrome and recent meta-analyses have supported their efficacy in this disease. Due to its therapeutic index, the use of n-butyl scopolammonium is more indicated to treat acute colics than a chronic disease such as irritable bowel syndrome. Taking into consideration the published trials carried out with validated methodology in irritable bowel syndrome, cimetropium and otilonium are the best demonstrated drugs for the improvement in global assessment, pain and abdominal distension.

Otilonium bromide inhibits calcium entry through L-type calcium channels in human intestinal smooth muscle.

Otilonium bromide (OB) is used as an intestinal antispasmodic. The mechanism of action of OB is not completely understood. As Ca(2+) entry into intestinal smooth muscle is required to trigger contractile activity, our hypothesis was that OB blocked Ca(2+) entry through L-type Ca(2+) channels. Our aim was to determine the effects of OB on Ca(2+), Na(+) and K(+) ion channels in human jejunal circular smooth muscle cells and on L-type Ca(2+) channels expressed heterologously in HEK293 cells. Whole cell currents were recorded using standard patch clamp techniques. Otilonium bromide (0.09-9 micromol L(-1)) was used as this reproduced clinical intracellular concentrations. In human circular smooth muscle cells, OB inhibited L-type Ca(2+) current by 25% at 0.9 micromol L(-1) and 90% at 9 micromol L(-1). Otilonium bromide had no effect on Na(+) or K(+) currents. In HEK293 cells, 1 micromol L(-1) OB significantly inhibited the expressed L-type Ca(2+) channels. Truncation of the alpha(1C) subunit C and N termini did not block the inhibitory effects of OB. Otilonium bromide inhibited Ca(2+) entry through L-type Ca(2+) at concentrations similar to intestinal tissue levels. This effect may underlie the observed muscle relaxant effects of the drug.