Update on experimental acute pancreatitis.

Acute pancreatitis is a disease still not fully understood. Early pathophysiologic event escape clinical observation because patients typically present only some time after the acute onset of the disease. Also, many ethiologic factors can lead to acute pancreatitis and the clinical course can range from mild, self-limiting to severe and life- threatening. Therefore, experimental models are necessary for any research into early acute pancreatitis. In accordance with the varying clinical picture of acute pancreatitis, many different model exist. In this article, we describe the most commonly used models and show their advantages and disadvantages.

Immunomodulation in a novel model of experimental chronic pancreatitis.

Chronic pancreatitis is a disease that involves the lymphocytic inflammation of the pancreatic gland, the destruction and fibrous transformation of the endocrine and ductal structures. An involvement of the immune system in the disease progression is assumed and possibly allows immune modulation as a novel treatment strategy. We used a new model of experimental chronic pancreatitis to examine the effect of immune modulation with the mTOR-inhibitor rapamycin on clinical, chemical and histological parameters of chronic pancreatitis. Pancreatitis was induced by injecting 8 mg/kg bodyweight DBTC intravenously in male Sprague Dawley rats. 24 and 72 hours later, 20 µg/kg bodyweight cerulein was injected intraperitoneally to simulate recurrent attacks of pancreatitis typical for the clinical course. 48 hours after the DBTC injection, rats were randomly allocated to placebo or sirolimus (1.5 mg/kg bw i.p.). The treatment was repeated every 24hours for 5 days. The rats were sacrificed 7, 14, 21 and 35 days after DBTC injection. Histologic examination revealed a reduced acute pancreatic damage in the treatment group in the first week and less chronic changes in the further course. ALT and amylase increased in Placebo animals over the observation period and was lower in sirolimus treated animals. Oral glucose tolerance test showed that all placebo animals were diabetic four weeks after DBTC while sirolimus treated animals were normoglycemic. An early, limited treatment with immunomodulatory and antifibrotic agents like sirolimus can positively influence the detrimental course of experimental chronic pancreatitis and may offer a treatment alternative in humans.

Is there a need for postoperative monitoring after open carpal tunnel release under WALANT?

Open carpal tunnel release (OCTR) under wide-awake local anesthesia with no tourniquet (WALANT) is a common outpatient procedure in hand surgery worldwide. In our clinic, WALANT has replaced intravenous regional anesthesia with a tourniquet (IVRA, or 'Bier block') as standard practice in OCTR. We therefore wondered what the optimal postoperative setting after OCTR under WALANT is. In this study, we compared patient satisfaction in two postoperative settings: immediate discharge (ID) after the operation, or short postoperative monitoring (PM) period in the outpatient clinic. Our hypothesis was that older patients would prefer a brief postoperative surveillance. We retrospectively analyzed patient satisfaction with the two settings using an adjusted questionnaire based on the standard Swiss grading system. We also assessed postoperative pain, satisfaction with the perioperative preparations and the reasons for unscheduled postoperative consultations, as secondary outcomes. One hundred and nine patients (ID, n = 63; PM, n = 46) were included in this single-center retrospective observational study. Patients were highly satisfied with both postoperative settings (Mean: ID 5.1/6; PM 5.5/6; p = 0.07). Even patients aged ≥80 years reported extremely high satisfaction with both settings (ID 5.6/6; PM 6.0/6; p = 0.08). Fifteen patients (ID, n = 11 [17.5%]; PM, n = 4 [8.7%], p = 0.72) unexpectedly consulted a doctor after surgery. OCTR under WALANT as an outpatient procedure with immediate discharge was associated with high patient satisfaction. However, detailed postoperative monitoring could contribute to the patient's well-being and education on how to cope with the postoperative course, and help with any questions.

Understanding C-H bond oxidations: H. and H- transfer in the oxidation of toluene by permanganate.

The oxidation of toluene by permanganate has been studied as a model for the oxidation of C-H bonds by metal reagents, metalloenzymes, and metal oxide surfaces. In water, the reaction proceeds by hydride (H-) transfer from toluene to a permanganate oxygen, whereas in toluene solution, permanganate abstracts a hydrogen atom (H.). The ability of permanganate to abstract a hydrogen atom is rationalized on the basis of the strong O-H bond formed on H. addition to permanganate. This approach allows understanding and prediction of the rates of hydrogen atom transfer from substrates to metal active sites.

Application of the Marcus cross relation to hydrogen atom transfer reactions.

The transfer of a hydrogen atom-a proton and an electron-is a fundamental process in chemistry and biology. A variety of hydrogen atom transfer reactions, involving iron complexes, phenols, hydroxylamines, tBuOOH, toluene, and related radicals, are shown to follow the Marcus cross relation. Thus, the Marcus theory formalism based on ground-state energetics and self-exchange rates, originally developed for electron transfer processes, is also valuable for hydrogen atom transfer. Compounds that undergo slow proton transfer (C-H bonds) or slow electron transfer (cobalt complexes) also undergo slow hydrogen atom transfer. Limitations of this approach are also discussed.

Everolimus and mycophenolate mofetil sensitize human pancreatic cancer cells to gemcitabine in vitro: a novel adjunct to standard chemotherapy?

BACKGROUND/AIMS: Gemcitabine improves survival in pancreatic adenocarcinoma. A variety of drugs have been tested to potentiate gemcitabine treatment for pancreatic cancer cells. Two major immunosuppressive drugs, mycophenolate mofetil (MMF) and everolimus (RAD001) have been shown to exert an anti-tumoral effect, but their ability to sensitize human pancreatic cell lines during gemcitabine treatment remains unclear. We examined the effects of everolimus and MMF on gemcitabine-treated MiaPaCa and Panc-1 cell lines. METHODS: MiaPaCa and Panc-1 human pancreatic tumor cell lines were subjected to everolimus (0.001-1 microg/ml) or MMF (0.1-100 microg/ml) treatment in combination with gemcitabine (1-10(6) nM). Western blot analysis was performed for Panc-1 cells in the presence or absence of TGF-beta1 and different treatments: 0.1-100 muicro/ml MMF and 0.1-100 microg/ml everolimus. The antiproliferative effect of the treatment was assessed by BrdU test. The results were evaluated by two-way analysis of variance followed by post-hoc tests, and nonlinear regression analysis for dose-response rates. RESULTS: As expected, standard treatment doses of gemcitabine decreased proliferation dose-dependently. Everolimus increased the actual EC(50) response to gemcitabine treatment (1-10(3) nM) to as much as 83.1 and 82.1% in MiaPaCa and Panc-1 cell lines, respectively. Likewise, concomitant administration with MMF altered the EC(50) of gemcitabine treatment in MiaPaCa cell lines to values between 76.8 and 85.2% for doses of >or=1 microg/ml. Even the minor dose of MMF (0.1 microg/ml) increased the antiproliferative effect of gemcitabine by 43.5% for MiaPaCa and 42.4% for Panc-1 cells. In addition, treatment of Panc-1 cells with MMF (0.1-100 microg/ml) dose-dependently inhibited TGF-beta1-induced collagen expression. CONCLUSION: We found an overadditive antiproliferative effect of both MMF and everolimus in gemcitabine-treated MiaPaCa and Panc-1 cells in vitro, and an additional inhibitory effect of MMF on TGF-beta1-induced collagen type I expression. Interestingly, both the sensitizing effect of pancreatic cancer cells to gemcitabine treatment and the inhibition of collagen type I expression could be achieved by clinically feasible doses of everolimus and MMF. The use of these drugs is promising as a novel adjunct to standard chemotherapy.

Antiproliferative and overadditive effects of everolimus and mycophenolate mofetil in pancreas and lung cancer cells in vitro.

BACKGROUND: Everolimus inhibits the growth of several tumor cell lines in vitro as well as tumor growth in a rat model. Mycophenolate mofetil (MMF) inhibits growth of a Walker sarcoma in a rat model in vivo. Herein we tested the in vitro antiproliferative capacity of everolimus and MMF on a pancreatic tumor cell line Panc-1 and on a small cell lung cancer cell line ScLc. MATERIALS AND METHODS: Cells were cultured under standardized conditions. Everolimus was added in increasing doses from 0.005 to 500 microg/mL; MMF was used from 0.05 to 5000 microg/mL. For co-incubation experiments, we combined everolimus (0.005 microg/mL and 0.05 microg/mL) with five concentrations of MMF; and MMF (0.5 microg/mL and 5 microg/mL) with five concentrations of everolimus. The antiproliferative capacity was assessed by a BrdU incorporation assay. RESULTS: Everolimus and MMF inhibited BrdU incorporation into Panc-1 and ScLc in a dose-dependent fashion. A 50% inhibition was seen in Panc-1 only at 50 microg/mL everolimus, but in ScLc at 5 microg/mL everolimus. MMF was clearly more potent in Panc-1: 50% inhibition was observed at 5 microg/L. In ScLc, 40% inhibition of BrdU incorporation was seen only at 50 microg/L MMF. In co-incubation, an effective combination for both Panc-1 and ScLc was 5 microg/mL MMF with 0.005 microg/mL everolimus resulting in 50% inhibition of BrdU incorporation (P < .001). CONCLUSIONS: Everolimus and MMF showed dose-dependent antiproliferative effects in tumor cell lines in vitro both alone and in combination. The combined use of everolimus and MMF showed supra-additive effects at concentrations used for therapeutic immunosuppression in patients.

Pseudo-thermosetting chitosan hydrogels for biomedical application.

To prepare transparent chitosan/beta-glycerophosphate (betaGP) pseudo-thermosetting hydrogels, the deacetylation degree (DD) of chitosan has been modified by reacetylation with acetic anhydride. Two methods (I and II) of reacetylation have been compared and have shown that the use of previously filtered chitosan, dilution of acetic anhydride and reduction of temperature in method II improves efficiency and reproducibility. Chitosans with DD ranging from 35.0 to 83.2% have been prepared according to method II under homogeneous and non-homogeneous reacetylation conditions and the turbidity of chitosan/betaGP hydrogels containing homogeneously or non-homogeneously reacetylated chitosan has been investigated. Turbidity is shown to be modulated by the DD of chitosan and by the homogeneity of the medium during reacetylation, which influences the distribution mode of the chitosan monomers. The preparation of transparent chitosan/betaGP hydrogels requires a homogeneously reacetylated chitosan with a DD between 35 and 50%.

Pseudo-thermosetting chitosan hydrogels for biomedical application.

To prepare transparent chitosan/beta-glycerophosphate (betaGP) pseudo-thermosetting hydrogels, the deacetylation degree (DD) of chitosan has been modified by reacetylation with acetic anhydride. Two methods (I and II) of reacetylation have been compared and have shown that the use of previously filtered chitosan, dilution of acetic anhydride and reduction of temperature in method II improves efficiency and reproducibility. Chitosans with DD ranging from 35.0 to 83.2% have been prepared according to method II under homogeneous and non-homogeneous reacetylation conditions and the turbidity of chitosan/betaGP hydrogels containing homogeneously or non-homogeneously reacetylated chitosan has been investigated. Turbidity is shown to be modulated by the DD of chitosan and by the homogeneity of the medium during reacetylation, which influences the distribution mode of the chitosan monomers. The preparation of transparent chitosan/betaGP hydrogels requires a homogeneously reacetylated chitosan with a DD between 35 and 50%.

Influence of palmitate and benzoate on the unidirectional chiral inversion of ibuprofen in isolated rat hepatocytes.

The influence of benzoic acid, a typical substrate of medium-chain acyl-CoA synthetase, and of palmitic acid, a substrate of long-chain acyl-CoA synthetase, on the metabolic chiral inversion of ibuprofen was investigated in freshly isolated hepatocytes. It was shown that the conjugation of benzoid to hippuric acid does not influence the chiral inversion of ibuprofen. In contrast, palmitic acid inhibited markedly the R-to-S inversion of ibuprofen. It was concluded that this inhibition is due to competition between (R)-ibuprofen and palmitic acid for long-chain acyl-CoA synthetases.

Dual ligand binding of pyridylalkanamides to microsomal cytochrome P-450.

Twelve homologous and regioisomeric pyridylalkanamides were examined spectrally for their binding affinity to cytochrome P-450 in phenobarbital- and 3-methylcholanthrene-induced rat liver microsomes. The pKs values were calculated by the Lineweaver-Burk method and by non-linear analysis using both a one ligand-one acceptor and a one ligand-two acceptor model. The latter model best fits most of the data, confirming that two pKs values exist for most derivatives in the 3-pyridyl and 4-pyridyl series. Structure-binding relationships are discussed. The two binding constants are hypothesized to arise from a dual mode of binding to the ferric ion. At low ligand concentrations, binding to hexacoordinated cytochrome P-450 occurs and involves displacement of an endogenous 6th ligand; at higher concentrations, the ligands bind to the pentacoordinated P-450, resulting in a high-to-low spin shift.

Development of quantitative structure-pharmacokinetic relationships.

Quantitative structure-activity relationships (QSAR) relating biological activity to physiochemical descriptors have been successfully used for a number of years. It is also long recognized that pharmacokinetic parameters may play an important and even determinant role in drug action. This prompted several researchers to focus attention to pharmacokinetic parameters as potential descriptors in quantitative drug design. A number of examples of quantitative structure-pharmacokinetic relationships (QSPR) have appeared in the literature. The present contribution reviews some developments in this field. In particular, a number of concepts and problems are critically discussed, rather than compilations of examples already published in recent reviews. Attention will be paid to the main processes of the pharmacokinetic or toxicokinetic phase in drug action, including absorption, distribution and elimination (biotransformation and excretion). It is clear that quantitative approaches are of considerable interest to toxicologists, since these methods may contribute to the development of real predictive toxicology.

Differential pharmacological responses to ethanol, pentobarbital and morphine in rats selectively bred for ethanol sensitivity.

The hypothermic and analgesic effects of ethanol, pentobarbital and morphine were examined in two lines of rats that had been selectively bred for their differential sensitivity to ethanol. Males and females of the least-affected (LA) line were observed to be less sensitive than their most-affected (MA) counterparts to hypothermia and analgesia induced by ethanol and morphine. By contrast, no differences were observed with respect to pentobarbital-induced hypothermia. At the dose used, pentobarbital had no significant analgesic effect in either animal line.

Factors involved in the differential response to ethanol, barbital and pentobarbital in rats selectively bred for ethanol sensitivity.

Two lines of rats, 'least affected' (LA) and 'most affected' (MA), had been selectively bred for their differential sensitivity to ethanol. Both males and females of the LA strain were observed to be less sensitive than their MA counterparts to the acute hypnotic and motor-impairing effects of ethanol. However, a lower ethanol metabolic rate of the MA males suggests that both CNS and metabolic factors contribute to their enhanced sensitivity to ethanol. By contrast, no differences were observed between the LA and MA males with respect to the hypnotic and subhypnotic effects of pentobarbital or to the clearance of this drug. MA females were more sensitive only to the hypnotic effects of pentobarbital, probably because of a smaller apparent volume of distribution. No strain difference was observed in the hypnotic effect or clearance of barbital. These observations suggest that, in spite of a differential sensitivity to ethanol, the LA and MA lines do not differ in their response to the barbiturates tested.

Ibuprofen enantiomers and lipid metabolism.

Recent findings concerning the mechanism of the chiral inversion of "profens" have given a better understanding of the ways in which profens interact with lipid biochemistry. This study presents investigations and findings concerning the influence of coenzyme-A (CoA) levels on the chiral inversion of ibuprofen. Measurement of intracellular coenzyme-A levels in isolated rat hepatocytes revealed that R-ibuprofen transiently reduced coenzyme-A levels, whereas S-ibuprofen had no effect. Other experiments were performed with rat hepatocyte suspensions, including tests with various concentrations of clofibric acid added to incubates. Results showed that both clofibric acid pretreatment and its presence in the perfusion medium increases the chiral inversion of R-ibuprofen. These results confirm a metabolic interaction between ibuprofen and clofibric acid. Clinical studies are continuing to determine whether this metabolic interaction has toxicologic consequences.

A role for calcium in the acute and chronic actions of ethanol in vitro.

Ethanol, in a concentration of 40--160 mM, depressed the electrically evoked contractions of the guinea-pig ileum longitudinal muscle/myenteric plexus (LM/MP) preparation in a dose-dependent manner. The acute ethanol inhibition in vitro was partially reversed by raising the extracellular calcium concentration. Following a 4 week ethanol treatment in vivo, by means of daily i.p. injections, the LM/MP preparation was rendered tolerant to ethanol and at the same time calcium was more effective in reversing the inhibitory effect of ethanol. It appears that calcium plays a role in the acute action of, and development of tolerance to, ethanol.

Different sites of action for morphine, ethanol, barbital and pentobarbital within the guinea-pig ileum longitudinal muscle/myenteric plexus preparation.

Morphine, ethanol, barbital and pentobarbital reversibly inhibited in a dose-dependent manner, the electrically evoked contractions of the guinea-pig ileum longitudinal muscle/myenteric plexus (LM/MP) preparation, in vitro. The ED50s for inhibition were 0.2 microM for morphine and 120, 8 and 0.3 mM for ethanol, barbital and pentobarbital respectively. To determine whether the site of inhibition was pre- or postjunctioal we studied the effects of graded concentrations of these drugs on the contractions evoked by exogenous acetylcholine (ACh). A correlation between the inhibition of the electrically and ACh-evoked muscle contractions suggests that morphine acts primarily prejunctionally while ethanol acts both pre-and postjunctionally. The inhibition by barbital and pentobarbital, however, can be accounted completely by a postjunctional depression. These results may explain our previous observation of cross-tolerance between morphine and ethanol in this preparation, and the lack of cross-tolerance between these drugs and barbital.

Cross-tolerance between ethanol and morphine in the guinea-pig ileum longitudinal-muscle/myenteric-plexus preparation.

Morphinization of guinea pigs with s.c. implanted pellets (4 X 75 mg morphine base) has resulted in tolerance, within the isolated longitudinal muscle/myenteric plexus preparation, to the in vitro inhibitory effects of morphine on the electrically induced contractions, and in cross-tolerance to the inhibitory effects of ethanol. Similar results were observed with opiate-naive preparations that were incubated for 18 h in Krebs-Ringer solution containing 40 muM morphine. Daily ethanol injections (6 g/kg) resulted in tolerance to ethanol and cross-tolerance to morphine. These results suggest a commonality between ethanol and morphine actions on the myenteric plexus preparation.

Familial juvenile parkinsonism with multiple systems degenerations. A clinicopathological study.

An unusual case of familial multisystemic degeneration is reported. Two siblings had juvenile parkinsonism, areflexia, and retinal degeneration of slow progression. The main neuropathological findings in case 1 were pallidoluysian, nigral, dentate, and dorsal columns degeneration. The authors draw a comparison between this case and juvenile parkinsonism, dentato-rubro-pallido-luysian atrophy, and spino-cerebello-nigral degeneration.

Structure and interactions in chitosan hydrogels formed by complexation or aggregation for biomedical applications.

The aim of this review was to provide a detailed overview of physical chitosan hydrogels and related networks formed by aggregation or complexation, which are intended for biomedical applications. The structural basis of these systems is discussed with particular emphasis on the network-forming interactions, the principles governing their formation and their physicochemical properties. An earlier review discussing crosslinked chitosan hydrogels highlighted the potential negative influence on biocompatibility of covalent crosslinkers and emphasised the need for alternative hydrogel systems. A possible means to avoid the use of covalent crosslinkers is to prepare physical chitosan hydrogels by direct interactions between polymeric chains, i.e. by complexation, e.g. polyelectrolyte complexes (PEC) and chitosan/poly (vinyl alcohol) (PVA) complexes, or by aggregation, e.g. grafted chitosan hydrogels. PEC exhibit a higher swelling sensitivity towards pH changes compared to covalently crosslinked chitosan hydrogels, which extends their potential application. Certain complexed polymers, such as glycosaminoglycans, can exhibit interesting intrinsic properties. Since PEC are formed by non-permanent networks, dissolution can occur. Chitosan/PVA complexes represent an interesting alternative for preparing biocompatible drug delivery systems if pH-controlled release is n/ot required. Grafted chitosan hydrogels are more complex to prepare and do not always improve biocompatibility compared to covalently crosslinked hydrogels, but can enhance certain intrinsic properties of chitosan such as bacteriostatic and wound-healing activity.