Cardiopulmonary effects of buprenorphine in horses

Objective - To investigate the effects of buprenorphine on cardiopulmonary variables and on abdominal auscultation scores in horses. Animals - 6 healthy adult horses. Procedures - Horses were restrained in stocks and allocated to 2 treatments in a randomized crossover design, with 1-week intervals between each treatment. Saline (0.9% NaCl) solution was administered IV as a control, whereas buprenorphine (10 μg/kg, IV) was administered to the experimental group. Cardiopulmonary data were collected for 120 minutes after buprenorphine or saline solution administration. Abdominal auscultation scores were monitored for 2 and 12 hours after drug administration in the control and experimental groups, respectively. Results - Following control treatment, horses remained calm while restrained in the stocks and no significant changes in cardiopulmonary variables were observed throughout the study. Buprenorphine administration caused excitatory phenomena (restlessness and head shaking). Heart rate, cardiac index, and arterial blood pressure were significantly increased after buprenorphine administration until the end of the observational period (120 minutes). Minimal changes were found in arterial blood gas tensions. Abdominal auscultation scores decreased significantly from baseline for 4 hours after buprenorphine administration. Conclusions and Clinical Relevance - Buprenorphine induced excitement and hemodynamic stimulation with minimal changes in arterial blood gas tensions. These effects may impact the clinical use of buprenorphine in horses. Further studies are indicated to investigate the effects of buprenorphine on gastrointestinal motility and fecal output.

Magnetic multiparticulate colonic delivery systems evaluated by AC Biosusceptometry

The biomagnetic technique called Alternate Current Biosusceptometry (ACB) is a proposal to evaluate a multiparticulate drug delivery system in the human gastrointestinal tract. Results show that ACB was able to quantify the gastrointestinal transit and spreading of the magnetic material and is an attractive tool for pharmaceutical research. © 2007.

Estudo comparativo entre a atropina e a hioscina na reversão da bradicardia induzida pela detomidina em equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da proteção gástrica e duodenal do monoterpeno nerol em roedores

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Farmacomagnetografia colônia: estudo in vitro da desintegração de comprimidos magnéticos revestidos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo da motilidade gástrica em ratas prenhes diabéticas e não diabéticas

Several hormonal and physiological changes that occur during pregnancy directly affect gastrointestinal motility (GI), however, very little is known about the relationship between the electrical and mechanical activities of the stomach and its implications. The aim of this study was to compare the female sex hormones, with profiles of gastrointestinal motility using AC biosusceptometry (BAC) in pregnant rats. Seven female Wistar rats (weighing 250-350 g) were used and all procedures were approved by the local Ethics Committee of animal. Laparotomy was performed on anesthetized rats and a magnetic marker (3.5 mm in diameter and 3.0 mm height) was implanted in the distal stomach. After 10 days of recuperation, the mice were put to mate until confirmation of pregnancy. Fed animals were anesthetized (mg pentobarbital 30 / kg) and kept supine for recording the signal with a sensor 30 min BAC placed on the abdominal surface on days 0, 7, 14 and 20 of gestation. Statistical analysis was performed by Student's t test statistical Newman-Keuls and a P value of .05 was considered statistically significant. The frequency of contraction of the stomach was 74 mHz ± 3, 67 ± 7 mHz, 59 mHz ± 6 (p <0.05 vs day -1) and 71 ± 5 mHz on days -0, -7, -14, and -20, respectively. The values of 44 ± 15 pg / mL and 31 ± 9 pg / ml 25 ± 6 pg / ml, 22 ± 9 pg / ml on days -0, -7, -14 and -20, respectively, obtained by 17ß- estradiol were not statistically significant. The values of progesterone were 13 ± 6 pg / ml at 54 ± 15 pg / ml, 127 ± 42 pg / ml (p <0.05 vs day-1), 22 ± 13 pg / mL on day -0, - 7, -14 and -20, respectively. A linear correlation (Pearson's test) between the lower frequency gastric and increased levels of progesterone was 80% (p <0.0000001). Until then, the relationship between changes in motility and hormone were not described in detail, but with the work that we were able to demonstrate the high correlation between progesterone concentration ...

Tomografia por Biosusceptometria de Corrente Alternada (tBAC): desenvolvimento, caracterização, implementação

The Biosusceptometry AC (BAC) is a research tool that has been extensively explored by the group Biomagnetism IBB-UNESP for monitoring of the gastrointestinal tract, its response to a known drug or in vivo performance of solid dosage forms. During this period the BAC, which has the characteristics of high sensitivity and low cost, has been developed primarily for recording signals contraction of activity and traffic human gastrointestinal tract. With the possibility of producing images with this instrumentation, it was possible to evaluate different situations in vitro and in vivo for physiological studies and pharmaceuticals. Considering the good performance of this system to produce planar images, the first aim of the BAC system tomography (TBAC) was to evaluate the system performance of BAC to produce tomographic images of phantoms ferromagnetic for a single channel system. All these applications were only possible because of their sensitivity to materials of high magnetic suscepitibility as ferrite, which allow to produce an electrical signal proportional to the variation of the magnetic flux generated by the presence of magnetic marker next to a first-order gradiometer. Measuring this variation at various points was possible to generate planar images that recently came to be produced in systems with multiple detectors, said multi-channels. From planar images, also producing tomographic images of simulators BAC bars in a system of 13 channels using only the center channel, with good results when applied to simple objects as one and two bars. When testing the resolution of the system with more elaborate forms the quality and resolution of images reconstructed is not satisfactory, which would be solved by increasing the spatial sampling rate and hence the acquisition time. The present system works with an acquisition time of about five hours. Whereas this system will be applied for in vivo experiments, the acquisition time became a ...

Avaliação de parâmetros farmacêuticos em comprimidos matriciais empregando a técnica de Biosusceptometria AC

Oral administration is widely accepted route for drug delivery and solid dosage forms are commonly employed. The variation of absorption profiles along the human gastrointestinal tract (GIT) and the ability to target drugs by adequate dosage forms to distinct sites is the challenge in the pharmaceutical development of solid dosage forms. AC Biosusceptometry (ACB) is a technique that deserves consideration due to its features, accuracy of results and versatility. The purpose of this work was to evaluate, by employing the AC Biosusceptometer, the rate of swelling of systems matrices consisting of hydrophilic polymer (hydroxypropyl methyl cellulose) and magnetic material. Matrices tablets were evaluated in vitro to a more detailed analysis of kinetics of swelling, in addition to the study and application of mathematical models to correlate the magnetic area variation and the water uptake. All the procedures for qualitative and quantitative analysis of digital signals as well as the magnetic images processing were performed in MatLab® (Mathworks Inc.). ACB technique proved to be useful towards estimating the swelling properties of hydrophilic matrices in vitro, showing a promising capacity for further analyses involving dissolution test and in vivo studies, supporting their innovative potential pharmaceutical applications

Testes preliminares de motilidade intestinal e toxicidade oral aguda com extrato de cascas pulverizadas de Endopleura uchi (Huber) Cuatrec: (Humiriaceae) em camundongos

The aim of this work was to analyze the acute oral toxicity and the effects on intestinal motility of the extract obtained through decoction 20% (m/v) of Endopleura uchi, popularly known as "uxi-amarelo", a native plant from the Brazilian Amazon. The plant is used indiscriminately against several diseases: arthritis, cholesterol, diabetes, ulcers, myomas, and intestinal illnesses in general, among others. The present results show that there were no significant alterations in intestinal motility and that the extract does not present signs of toxicity, showing its safety for consumption purposes.

Cross-Talk tra Bifidobacterium e intestino umano: impatto sulle attività health promoting

Bifidobacteria constitute up to 3% of the total microbiota and represent one of the most important healthpromoting bacterial groups of the human intestinal microflora. The presence of Bifidobacterium in the human gastrointestinal tract has been directly related to several health-promoting activities; however, to date, no information about the specific mechanisms of interaction with the host is available. The first health-promoting activities studied in these job was the oxalate-degrading activity. Oxalic acid occurs extensively in nature and plays diverse roles, especially in pathological processes. Due to its highly oxidizing effects, hyper absorption or abnormal synthesis of oxalate can cause serious acute disorders in mammals and be lethal in extreme cases. Intestinal oxalate-degrading bacteria could therefore be pivotal in maintaining oxalate homeostasis, reducing the risk of kidney stone development. In this study, the oxalate-degrading activity of 14 bifidobacterial strains was measured by a capillary electrophoresis technique. The oxc gene, encoding oxalyl-CoA decarboxylase, a key enzyme in oxalate catabolism, was isolated by probing a genomic library of B. animalis subsp. lactis BI07, which was one of the most active strains in the preliminary screening. The genetic and transcriptional organization of oxc flanking regions was determined, unravelling the presence of other two independently transcribed open reading frames, potentially responsible for B. animalis subsp. lactis ability to degrade oxalate. Transcriptional analysis, using real-time quantitative reverse transcription PCR, revealed that these genes were highly induced in cells first adapted to subinhibitory concentrations of oxalate and then exposed to pH 4.5. Acidic conditions were also a prerequisite for a significant oxalate degradation rate, which dramatically increased in oxalate pre-adapted cells, as demonstrated in fermentation experiments with different pH-controlled batch cultures. These findings provide new insights in the characterization of oxalate-degrading probiotic bacteria and may support the use of B. animalis subsp. lactis as a promising adjunct for the prophylaxis and management of oxalate-related kidney disease. In order to provide some insight into the molecular mechanisms involved in the interaction with the host, in the second part of the job, we investigated whether Bifidobacterium was able to capture human plasminogen on the cell surface. The binding of human plasminogen to Bifidobacterium was dependent on lysine residues of surface protein receptors. By using a proteomic approach, we identified six putative plasminogen-binding proteins in the cell wall fraction of three strain of Bifidobacterium. The data suggest that plasminogen binding to Bifidobactrium is due to the concerted action of a number of proteins located on the bacterial cell surface, some of which are highly conserved cytoplasmic proteins which have other essential cellular functions. Our findings represent a step forward in understanding the mechanisms involved in the Bifidobacterium-host interaction. In these job w studied a new approach based on to MALDI-TOF MS to measure the interaction between entire bacterial cells and host molecular target. MALDI-TOF (Matrix Assisted Laser Desorption Ionization-Time of Flight)—mass spectrometry has been applied, for the first time, in the investigation of whole Bifidobacterium cells-host target proteins interaction. In particular, by means of this technique, a dose dependent human plasminogen-binding activity has been shown for Bifidobacterium. The involvement of lysine binding sites on the bacterial cell surface has been proved. The obtained result was found to be consistent with that from well-established standard methodologies, thus the proposed MALDI-TOF approach has the potential to enter as a fast alternative method in the field of biorecognition studies involving in bacterial cells and proteins of human origin.

[Drug therapy of acute and chronic abdominal pain]

For drug therapy a differentiation of acute and chronic pain is essential. In emergency situations of acute abdominal pain a fast diagnosis is mandatory. Analgesia should be provided as soon as possible. The different groups of analgesics should be used according to their known effects, side effects and contraindications. Postoperative pain after abdominal surgery has to be considered as a special condition of acute abdominal pain. Main treatment options are non opioid analgesics and opioids. Opioids can be administered intravenously via patient controlled analgesia (PCA) devices. In major abdominal surgery neuroaxial analgesia, preferentially administered via an epidural catheter provides excellent pain relief with positive impact on gastrointestinal motility and patients' recovery. Because of difficulties to allocate chronic abdominal pain to a specific organ, causal treatment often turns out to be difficult. Peripheral and central sensitization, as well as an alteration of the endogenous pain modulation comes to the fore in these chronic pain conditions. Co-analgesics like anticonvulsants and antidepressants are utilized to reduce sensitization and improve the endogenous pain modulating system. Non drug approaches and alternative treatment options might be useful. In contrast, orally or transcutaneously administered opioids are the principal corner stone for the treatment of cancer pain.

PCR and real-time PCR primers developed for detection and identification of Bifidobacterium thermophilum in faeces

BACKGROUND: Culture-independent methods based on the 16S ribosomal RNA molecule are nowadays widely used for assessment of the composition of the intestinal microbiota, in relation to host health or probiotic efficacy. Because Bifidobacterium thermophilum was only recently isolated from human faeces until now, no specific real-time PCR (qPCR) assay has been developed for detection of this species as component of the bifidobacterial community of the human intestinal flora. RESULTS: Design of specific primers and probe was achieved based on comparison of 108 published bifidobacterial 16S rDNA sequences with the recently published sequence of the human faecal isolate B. thermophilum RBL67. Specificity of the primer was tested in silico by similarity search against the sequence database and confirmed experimentally by PCR amplification on 17 Bifidobacterium strains, representing 12 different species, and two Lactobacillus strains. The qPCR assay developed was linear for B. thermophilum RBL67 DNA quantities ranging from 0.02 ng/microl to 200 ng/microl and showed a detection limit of 10(5) cells per gram faeces. The application of this new qPCR assay allowed to detect the presence of B. thermophilum in one sample from a 6-month old breast-fed baby among 17 human faecal samples tested. Additionally, the specific qPCR primers in combination with selective plating experiments led to the isolation of F9K9, a faecal isolate from a 4-month old breast-fed baby. The 16S rDNA sequence of this isolate is 99.93% similar to that of B. thermophilum RBL67 and confirmed the applicability of the new qPCR assay in faecal samples. CONCLUSION: A new B. thermophilum-specific qPCR assay was developed based on species-specific target nucleotides in the 16S rDNA. It can be used to further characterize the composition of the bifidobacterial community in the human gastrointestinal tract. Until recently, B. thermophilum was considered as a species of animal origin, but here we confirm with the application of this new PCR assay the presence of B. thermophilum strains in the human gut.

Arginine vasopressin in vasodilatory shock: effects on metabolism and beyond

PURPOSE OF REVIEW: To describe the effects of arginine vasopressin other than its vasoconstrictive and antidiuretic potential in vasodilatory shock. RECENT FINDINGS: Arginine vasopressin influences substrate metabolism by stimulation of hepatic glucose release, gluconeogenesis, ureogenesis and fatty acid esterification. Although arginine vasopressin is a secretagogue of different hormones, only prolactin increases during arginine vasopressin therapy. Plasmatic and cellular coagulation are affected by arginine vasopressin, resulting in thrombocyte aggregation. Therefore, platelet count typically decreases following arginine vasopressin infusion in critically ill patients. In addition, arginine vasopressin reduces bile flow and may increase bilirubin concentrations. Despite its potential to decrease serum sodium, no change in electrolytes was observed in critically ill patients receiving arginine vasopressin. Although arginine vasopressin is an endogenous antipyretic, body temperature is not decreased by central venous arginine vasopressin infusion. In addition, arginine vasopressin modulates immune function through V1 receptors. Compared with norepinephrine, arginine vasopressin may have protective effects on endothelial function. Net arginine vasopressin effects on gastrointestinal motility seem to be inhibitory and are dose dependent. SUMMARY: Except for its antidiuretic and vasoconstrictive actions, the effects of arginine vasopressin in patients with vasodilatory shock have so far only been partially examined. Potential influences of arginine vasopressin on metabolism and immune, liver and mitochondrial function remain to be assessed in future studies.

Reverse biochemistry: Use of macromolecular protease inhibitors to dissect complex biological processes and identify a membrane-type serine protease in epithelial cancer and normal tissue

Serine proteases of the chymotrypsin fold are of great interest because they provide detailed understanding of their enzymatic properties and their proposed role in a number of physiological and pathological processes. We have been developing the macromolecular inhibitor ecotin to be a “fold-specific” inhibitor that is selective for members of the chymotrypsin-fold class of proteases. Inhibition of protease activity through the use of wild-type and engineered ecotins results in inhibition of rat prostate differentiation and retardation of the growth of human PC-3 prostatic cancer tumors. In an effort to identify the proteases that may be involved in these processes, reverse transcription–PCR with PC-3 poly(A)+ mRNA was performed by using degenerate oligonucleotide primers. These primers were designed by using conserved protein sequences unique to chymotrypsin-fold serine proteases. Five proteases were identified: urokinase-type plasminogen activator, factor XII, protein C, trypsinogen IV, and a protease that we refer to as membrane-type serine protease 1 (MT-SP1). The cloning and characterization of the MT-SP1 cDNA shows that it encodes a mosaic protein that contains a transmembrane signal anchor, two CUB domains, four LDLR repeats, and a serine protease domain. Northern blotting shows broad expression of MT-SP1 in a variety of epithelial tissues with high levels of expression in the human gastrointestinal tract and the prostate. A His-tagged fusion of the MT-SP1 protease domain was expressed in Escherichia coli, purified, and autoactivated. Ecotin and variant ecotins are subnanomolar inhibitors of the MT-SP1 activated protease domain, suggesting a possible role for MT-SP1 in prostate differentiation and the growth of prostatic carcinomas.

Contrasting roles for Myc and Mad proteins in cellular growth and differentiation.

The positive effects of Myc on cellular growth and gene expression are antagonized by activities of another member of the Myc superfamily, Mad. Characterization of the mouse homolog of human mad on the structural level revealed that domains shown previously to be required in the human protein for anti-Myc repression, sequence-specific DNA-binding activity, and dimerization with its partner Max are highly conserved. Conservation is also evident on the biological level in that both human and mouse mad can antagonize the ability of c-myc to cooperate with ras in the malignant transformation of cultured cells. An analysis of c-myc and mad gene expression in the developing mouse showed contrasting patterns with respect to tissue distribution and developmental stage. Regional differences in expression were more striking on the cellular level, particularly in the mouse and human gastrointestinal system, wherein c-Myc protein was readily detected in immature proliferating cells at the base of the colonic crypts, while Mad protein distribution was restricted to the postmitotic differentiated cells in the apex of the crypts. An increasing gradient of Mad was also evident in the more differentiated subcorneal layers of the stratified squamous epithelium of the skin. Together, these observations support the view that both downregulation of Myc and accumulation of Mad may be necessary for progression of precursor cells to a growth-arrested, terminally differentiated state.