Systemic oxytocin increases glutamate efflux in the nucleus accumbens core of cocaine-experienced male and female rats but only increases dopamine efflux in males.

Oxytocin attenuates cocaine-seeking when administered both systemically and directly into the nucleus accumbens core. This effect is blocked by intra-accumbens antagonism of mGlu2/3 and, together with our finding that intra-accumbens oxytocin increases glutamate concentrations in this brain region, indicates that pre-synaptic regulation of glutamate release by oxytocin influences cocaine relapse. However, mGlu2/3 receptors also regulate dopamine release in the nucleus accumbens. Here we aimed to determine whether systemic oxytocin increases glutamate and dopamine concentrations in the nucleus accumbens core of cocaine-experienced and cocaine-naïve male and female rats. A subset of rats self-administered cocaine (0.5 mg/kg/infusion) and then underwent extinction training for 2-3 weeks. Rats were implanted with microdialysis probes in the accumbens core and samples were collected for a baseline period, and following saline (1 mL/kg), and oxytocin (1 mg/kg, IP) injections. Locomotion was assessed during microdialysis. In cocaine-experienced rats, oxytocin increased glutamate concentrations in the accumbens core to the same extent in males and females but only increased dopamine concentrations in male rats. Oxytocin did not alter glutamate levels in cocaine-naïve rats. Oxytocin did not produce sedation. These results extend previous findings that systemic oxytocin increases nucleus accumbens dopamine in a sex-specific manner in cocaine-experienced rats. These data are the first to find that systemic oxytocin increases nucleus accumbens glutamate after cocaine experience, providing a mechanism of action by which oxytocin attenuates the reinstatement of cocaine seeking in both male and female rats.

Effects of Methamphetamine Self-Administration and Extinction on Astrocyte Structure and Function in the Nucleus Accumbens Core.

Astrocytes provide support for neurons, regulate metabolic processes, and influence neuronal communication in a variety of ways, including through the homeostatic regulation of glutamate. Following 2-h cocaine or methamphetamine self-administration (SA) and extinction, rodents display decreased levels of basal glutamate in the nucleus accumbens core (NAcore), which transitions to elevated glutamate levels during drug seeking. We hypothesized that, like cocaine, this glutamate 'overflow' during methamphetamine seeking arises via decreased expression of the astroglial glutamate transporter GLT-1, and withdrawal of perisynaptic astroglial processes (PAPs) from synapses. As expected, methamphetamine self-administration and extinction decreased the level of contact made by PAPs in the NAcore, yet did not impact glutamate uptake, GLT-1 expression, or the general structural characteristics of astrocytes. Interestingly, systemic administration of N-acetylcysteine (NAC), a drug that both upregulates GLT-1 and promotes glial-glutamate release, reduced cued methamphetamine seeking. In order to test the impact of astrocyte activation and the induction of glial glutamate release within the NAcore, we employed astrocyte-specific expression of designer receptors exclusively activated by designer drugs (DREADDs). We show here that acute activation of Gq-coupled DREADDs in this region inhibited cued methamphetamine seeking. Taken together, these data indicate that cued methamphetamine seeking following two-hour SA is not mediated by deficient glutamate clearance in the NAcore, yet can be inhibited by engaging NAcore astrocytes.

[Recent developments in head and neck immunology : A basis for novel therapeutic strategies?] / Jüngste Entwicklungen in der Kopf-Hals-Immunologie : Grundlage für neue Therapiestrategien?

Immunological processes play a key role in the pathogenesis of head and neck pathologies. Besides allergies or infections of the tonsils, the paranasal sinuses, and the ear, initiation, progression, and metastasis of malignant tumors are particularly dependent on the immune system. The recruitment of white blood cells to the site of injury or infection is a critical event in the pathogenesis of these diseases. This article will provide a compact overview about recent developments in this rapidly growing field in otorhinolaryngology which might establish the basis for promising therapeutic strategies for previously insufficiently treatable disorders of the head and neck.

Factor VII-activating protease deficiency promotes neointima formation by enhancing leukocyte accumulation.

Essentials Factor VII-activating protease (FSAP) is a plasma protease involved in vascular processes. Neointima formation was investigated after vascular injury in FSAP-/- mice. The neointimal lesion size and the accumulation of macrophages were increased in FSAP-/- mice. This was due to an increased activity of the chemokine (C-C motif) ligand 2 (CCL2). SUMMARY: Background Factor VII-activating protease (FSAP) is a multifunctional circulating plasma serine protease involved in thrombosis and vascular remodeling processes. The Marburg I single-nucleotide polymorphism (MI-SNP) in the FSAP-coding gene is characterized by low proteolytic activity, and is associated with increased rates of stroke and carotid stenosis in humans. Objectives To determine whether neointima formation after vascular injury is increased in FSAP-/- mice. Methods and Results The neointimal lesion size and the proliferation of vascular smooth muscle cells (VSMCs) were significantly enhanced in FSAP-/- mice as compared with C57BL/6 control mice after wire-induced injury of the femoral artery. Accumulation of leukocytes and macrophages was increased within the lesions of FSAP-/- mice at day 3 and day 14. Quantitative zymography demonstrated enhanced activity of gelatinases/matrix metalloproteinase (MMP)-2 and MMP-9 within the neointimal lesions of FSAP-/- mice, and immunohistochemistry showed particular costaining of MMP-9 with accumulating leukocytes. Using intravital microscopy, we observed that FSAP deficiency promoted the intravascular adherence and the subsequent transmigration of leukocytes in vivo in response to chemokine ligand 2 (CCL2). CCL2 expression was increased in FSAP-/- monocytes but not in the vessel wall. There was no difference in the expression of platelet-derived growth factor (PDGF-BB). Conclusions FSAP deficiency causes an increase in CCL2 expression and CCL2-mediated infiltration of leukocytes into the injured vessel, thereby promoting SMC proliferation and migration by the activation of leukocyte-derived gelatinases. These results provide a possible explanation for the observed association of the loss-of-function MI-SNP with vascular proliferative diseases.

[Bilateral simultaneous central retinal vein occlusion in protein S deficiency]. / Bilateraler simultaner retinaler Zentralvenenverschluss bei Protein-S-Mangel.

CASE REPORT: This article reports a case of bilateral simultaneous central retinal vein occlusion (CRVO) and protein S deficiency. The 27-year-old male patient presented with a sudden decrease in vision in both eyes. The patient's medical history documented the death of his father at the age of 33 years due to pulmonary embolism. Thrombophilia screening revealed protein S deficiency. OBJECTIVES: We report this case to emphasize that in any case of young onset retinal vein occlusion, protein S deficiency should be suspected. CONCLUSIONS: Thrombophilia assays and taking a thorough medical history should be performed.

Optical magnification devices in tonsillectomy: a prospective randomised clinical study.

Tonsillectomy is one of the most common surgical procedure in otorhinolaryngology. A plethora of approaches has been undertaken so far to limit postoperative pain, one of the major problems patients are concerned with. Thermal damages of the surrounding tissue caused by coagulation during surgery are discussed to correlate with postoperative pain. Therefore, we studied whether the use of magnification devices reduced coagulation procedures and consequently limited post-operative pain. Following an intraindividual design, we performed tonsillectomy on one side using a microscope or magnifying glasses whereas the opposite side was operated with unsupported vision. As verified by a visual analogue scale, our study shows that neither the use of a microscope, nor the use of magnifying glasses leads to less post-operative pain. Other parameters like post-operative bleeding, duration of surgery, and total applied energy by bipolar coagulation were also comparable in the different treatment groups. Taken together, magnification-supported tonsillectomy does not seem to be appropriate for limiting complications of tonsillectomy, especially not for reducing post-operative pain.

Cathelicidin LL-37 in bronchoalveolar lavage and epithelial lining fluids from healthy individuals and sarcoidosis patients.

Sarcoidosis is a granulomatous disease of unknown etiology most often characterized by pulmonary manifestations. Changes in an innate immune system, involving antimicrobial peptides, have been noted during the course of pulmonary sarcoidosis. This study focuses on the level of LL-37 peptide, the only human cathelicidin, additionally characterized by a wide range of pleiotropic activities, in pulmonary sarcoidosis. A cross-sectional study was conducted in groups of 32 patients with sarcoidosis and 12 healthy individuals. Bronchoalveolar lavage fluid (BALF) sampling, followed by LL-37 measurements by mass spectrometry combined with previous immunoaffinity purification, was performed. Based on urea levels, concentrations of LL-37 in epithelial lining fluid (ELF) were calculated. The levels of LL-37 peptide in BALF samples derived from patients with pulmonary sarcoidosis (median: 17.45 pg/ml, 25th-75th percentile: 8.05-28.33 pg/ml) were significantly higher compared to the healthy group (median: 6.38 pg/ml, 25th-75th percentile: 4.90-11.55 pg/ml) (U Mann-Whitney test, p=0.04). Assessment of LL-37 in ELF confirmed the differences across the groups that were observed in BALF. The level of LL-37 in patients with sarcoidosis (median: 2.25 ng/ml, 25th-75th percentile: 1.03-5.06 ng/ml) was again higher compared to healthy individuals (median: 0.62 ng/ml, 25th-75th percentile: 0.43-2.17 ng/ml) (p=0.06, Mann-Whitney U test). The results of this study demonstrate that the level of LL-37 peptide is elevated in pulmonary compartment affected by sarcoidosis. This might have a meaning in the pathomechanism of the disease, especially taking into consideration versatile activity of human cathelicidin revealed in numerous experimental studies during the last years.

Sonography: the leading diagnostic tool for diseases of the salivary glands.

Ultrasound examination is the imaging procedure with the best predictive diagnostic capability for the salivary glands. Due to the salivary glands' relatively superficial anatomical location, clear boundary from surrounding tissue and comparatively typical echogenicity, therefore sonography is ideal for diagnosis. In addition, the technical advances in recent years, including higher resolution, color Doppler sonography, contrast-enhanced ultrasound, elastography, and tissue harmonic have lead to an improvement in diagnostic accuracy of sonography further resulting in an expansion of the range of indications. Sonography allows detection of obstructive salivary gland diseases such as stenosis or sialolithiasis, as well as sialadenosis such as Sjögren syndrome. Ultrasound examination alone is sufficient to diagnose benign tumors. However, in the case of malignant tumors, computer tomography or MRI may be also required, especially to determine the question of infiltration of the skull base.

Cathelicidin LL-37 in bronchoalveolar lavage and epithelial lining fluids from COPD patients and healthy individuals.

Innate immunity is currently under scope of interest concerning its role in the development of chronic obstructive pulmonary disease (COPD). Antimicrobial peptides constitute a potent part of this fast response system. Here, we focus on the role of a specific antimicrobial peptide, the only human cathelicidin, the pleiotropic LL-37 peptide, in the development of COPD under clinical conditions. A cross-sectional study was conducted in groups of 43 patients with COPD (previously classified according to GOLD) and 12 healthy individuals. Bronchoalveolar lavage fluid (BALF) sampling, followed by LL-37 measurements by mass spectrometry combined with previous immunoaffinity purification, was performed. Based on urea levels, concentrations of LL-37 in epithelial lining fluid (ELF) were calculated. Additionally, an antimicrobial assay of growth inhibition of two bacterial species, often involved in COPD development mechanisms, by purchased LL-37 was conducted. Altogether, 55 BALF samples were analyzed. LL-37 levels were significantly higher in BALF from patients in early stages of COPD (GOLD I-II) compared to BALFs from healthy individuals. The same was true for ELF. Cathelicidin’s concentration was significantly lower in both BALF and ELF from patients in advanced COPD (GOLD III-IV). The significantly elevated LL-37 levels both in BALF and ELF in patients with COPD at stage GOLD I-II together with reduced levels in advanced (COPD stage III-IV) further supports the innate immunity involvement in COPD pathology and suggests a profound change in non-specific immunity during the disease progression.

Quantum dots modulate leukocyte adhesion and transmigration depending on their surface modification.

Although different nanosized materials, including quantum dots (QDs), are intended to be used for biomedical applications, their interactions with microvessels and their inflammatory potential are largely unknown. In this in vivo study we report that leukocyte recruitment is modulated in the presence of quantum dots. We found that the surface chemistry of QDs strongly affects their localization in postcapillary venules, their uptake by perivascular macrophages, and their potential to modify steps of leukocyte recruitment.

Determination of nevirapine in plasma by GC-MS.

Here we describe a simple, stable, and specific gas chromatography-mass spectrometry (GC-MS) method for the determination of nevirapine in plasma. After precipitation of proteins, the non-nucleoside reverse transcriptase inhibitor nevirapine was extracted with dichloromethane. For the determination and quantification of nevirapine, 1 microL of the organic layer was injected onto the GC-MS system. Linear calibration curves were obtained with BIRH 0414BS as internal standard in a range from 0.01 to 15 microg/mL. Intra- and inter-day accuracy and precision of this method were good with an accuracy between 96-109% and a precision between 2-8% across the therapeutic range of nevirapine. GC-MS proved to be a valid alternative to high-performance liquid chromatography and liquid chromatography-MS.

[Significant changes of pharmacotherapy in gastroenterological rehabilitation of Crohn's disease]. / Signifikante Anderungen der Pharmakotherapie in der gastroenterologischen Rehabilitation von Patienten mit Morbus Crohn.

AIM: The pivotal role of optimizing pharmacotherapy is generally accepted in somatic rehabilitation of various specialities like cardiopulmonary rehabilitation. No data exist as to whether significant modifications of pharmacotherapy occur during gastroenterological rehabilitation of Crohn's Disease (CD) patients. METHODS: A single centre chart review was performed including patients with International Classification of Disease Codes for CD (ICD K50). The Harvey-Bradshaw activity index (HBI) and CD medications were protocolled at the beginning and end of in-patient rehabilitation. RESULTS: 337 of 355 patients with ICD K50 fulfilled the predefined diagnostic criteria of mild to moderate CD (250 female, 87 male, average age of 40 (95% confidenceinterval, 29-51)). Disease activity decreased from 4.9 to 3.7 by 1.2 (0.75-1.37) Units during 23 (20-35) days. On admission, 120 (36%) patients received one and 158 (47%) received two to five CD drugs. CD drug prescriptions changed in 162 (48%) patients. Overall, 116 (34%) patients received systemic steroids which were stopped in 14 patients (p<0.05). In the remaining 102 patients the cortisol equivalence doses decreased from 77 to 56 mg by 21 (14-28) mg. The number of patients on azathioprine (AZT) increased from 98 to 108 (p<0.05). The average AZT dose increased from 1.81 to 1.99 mg/kg in 97 rehabilitants continuously treated. CONCLUSION: Our results describe an association between rehabilitation and significant changes of CD-specific pharmacotherapy in line with current treatment guidelines. This supports the concept that future studies on effects of gastroenterological rehabilitation should control for changes in pharmacotherapy.

Postnatal manganese exposure alters dopamine transporter function in adult rats: Potential impact on nonassociative and associative processes.

In the present study, we examined whether exposing rats to a high-dose regimen of manganese chloride (Mn) during the postnatal period would depress presynaptic dopamine functioning and alter nonassociative and associative behaviors. To this end, rats were given oral supplements of Mn (750 microg/day) on postnatal days (PD) 1-21. On PD 90, dopamine transporter (DAT) immunoreactivity and [3H]dopamine uptake were assayed in the striatum and nucleus accumbens, while in vivo microdialysis was used to measure dopamine efflux in the same brain regions. The effects of postnatal Mn exposure on nigrostriatal functioning were evaluated by assessing rotorod performance and amphetamine-induced stereotypy in adulthood. In terms of associative processes, both cocaine-induced conditioned place preference (CPP) and sucrose-reinforced operant responding were examined. Results showed that postnatal Mn exposure caused persistent declines in DAT protein expression and [3H]dopamine uptake in the striatum and nucleus accumbens, as well as long-term reductions in striatal dopamine efflux. Rotorod performance did not differ according to exposure condition, however Mn-exposed rats did exhibit substantially more amphetamine-induced stereotypy than vehicle controls. Mn exposure did not alter performance on any aspect of the CPP task (preference, extinction, or reinstatement testing), nor did Mn affect progressive ratio responding (a measure of motivation). Interestingly, acquisition of a fixed ratio task was impaired in Mn-exposed rats, suggesting a deficit in procedural learning. In sum, these results indicate that postnatal Mn exposure causes persistent declines in various indices of presynaptic dopaminergic functioning. Mn-induced alterations in striatal functioning may have long-term impact on associative and nonassociative behavior.

[Differential diagnosis of jaundice]. / Virushepatitis oder Alkohol, Cholangitis oder Tumor? Was steckt hinter dem Symptom lkterus?

Jaundice is a symptom with a multitude of possible causes. These can be divided up into primary diseases of bilirubin metabolism, secondary hyperbilirubinemia in patients with liver disease, and diseases with bile duct occlusion. The major objective of the examination must be to exclude those causes that represent an acute danger to the patient, in particular cholangitis or cholecystitis. Symptoms that should cause alarm bells to ring include abdominal pain, fever and chills. When obtaining the patient's anamnesis, particular attention must be paid to lithiasis and previous operations. Laboratory findings of elevated alkaline phosphatase and gamma-GT indicate the presence of cholestasis. If an extrahepatic obstruction is suspected, ultrasonography of the upper abdomen is required.

Diagnostic accuracy of atypical p-ANCA in autoimmune hepatitis using ROC- and multivariate regression analysis.

INTRODUCTION: Antineutrophil cytoplasmic antibodies (atypical p-ANCA) are detected at high prevalence in sera from patients with autoimmune hepatitis (AIH), but their diagnostic relevance for AIH has not been systematically evaluated so far. METHODS: Here, we studied sera from 357 patients with autoimmune (autoimmune hepatitis n=175, primary sclerosing cholangitis (PSC) n=35, primary biliary cirrhosis n=45), non-autoimmune chronic liver disease (alcoholic liver cirrhosis n=62; chronic hepatitis C virus infection (HCV) n=21) or healthy controls (n=19) for the presence of various non-organ specific autoantibodies. Atypical p-ANCA, antinuclear antibodies (ANA), antibodies against smooth muscles (SMA), antibodies against liver/kidney microsomes (anti-Lkm1) and antimitochondrial antibodies (AMA) were detected by indirect immunofluorescence microscopy, antibodies against the M2 antigen (anti-M2), antibodies against soluble liver antigen (anti-SLA/LP) and anti-Lkm1 by using enzyme linked immunosorbent assays. To define the diagnostic precision of the autoantibodies, results of autoantibody testing were analyzed by receiver operating characteristics (ROC) and forward conditional logistic regression analysis. RESULTS: Atypical p-ANCA were detected at high prevalence in sera from patients with AIH (81%) and PSC (94%). ROC- and logistic regression analysis revealed atypical p-ANCA and SMA, but not ANA as significant diagnostic seromarkers for AIH (atypical p-ANCA: AUC 0.754+/-0.026, odds ratio [OR] 3.4; SMA: 0.652+/-0.028, OR 4.1). Atypical p-ANCA also emerged as the only diagnostically relevant seromarker for PSC (AUC 0.690+/-0.04, OR 3.4). None of the tested antibodies yielded a significant diagnostic accuracy for patients with alcoholic liver cirrhosis, HCV or healthy controls. CONCLUSIONS: Atypical p-ANCA along with SMA represent a seromarker with high diagnostic accuracy for AIH and should be explicitly considered in a revised version of the diagnostic score for AIH.

Diagnosis and therapy of intrahepatic cholestasis of pregnancy.

Intrahepatic cholestasis of pregnancy (ICP) is characterized by the occurrence of pruritus mostly in the third trimenon. Diagnosis is based on the presence of pruritus and elevated levels of serum bile acids in the absence of pruritic skin diseases. There is strong evidence of a genetic predisposition for ICP. Numerous studies have investigated the association of known cholestasis genes such as ABCB4 (also designated MDR3), ABCB11 ( BSEP) and ATP8B1 ( FIC1) with ICP. The results of these studies implicate a heterogeneous etiology of this syndrome. ICP increases the risk of preterm delivery and fetal loss. Furthermore, intense pruritus may necessitate premature induction of labor with its known higher frequency of complications for mother and child. Therefore, ICP pregnancies should be managed as high-risk pregnancies. Pharmaceuticals to alleviate pruritus or improve cholestasis like antihistamines, phenobarbital, anion exchange resins, dexamethasone or S-adenosylmethionine are not widely accepted because of questionable efficacy or side effects. Recent randomized studies have shown beneficial effects of ursodeoxycholic acid (UDCA) on laboratory data and pruritus in patients with ICP. Improved knowledge about the diagnostic classification of different types and pathophysiological mechanisms of ICP may allow for a more targeted treatment of this disease in future.

Protective effect of fructose-1,6-bisphosphate in the cold storage solution for liver preservation in rat hepatic transplantation.

Fructose-1,6-bisphosphate (FBP) has been reported to have a protective effect on liver injury following ischemic/reperfusion periods. FBP maintains ATP levels and thereby cellular energy metabolism, which is important to the liver during cold preservation. In the present study, we evaluated the effects of FBP on the composition of storage solutions for cold liver preservation. Adult male Wistar rats were randomly divided into three experimental groups. Hepatic perfusion and preservation were performed with UW, UW plus 10 mmol/L FBP (UWM), and FBP 10 mmol/L (FBPS) alone solutions. Biochemical measurements of AST, ALT, and TBARS were performed on samples of the cold storage solution at 0, 12, 18, and 24 hours preservation. FBPS and UW solutions showed similar preservation grades during 18 hours. Addition of 10 mmol/L of FBP to UW solution induced liver injury and a poor preservation grade. FBP appears to protect the liver from injury caused by free radicals when the preservation time is less than 18 hours. Therefore, FBP may exert a protective effect for the preservation of livers during cold storage, and could represent an important component of new cold storage solutions.

Influence of rifampin on serum markers of cholesterol and bile acid synthesis in men.

OBJECTIVE: It has been demonstrated in preliminary studies that rifampin, a semisynthetic antibiotic and known inducer of hepatic cytochrome P450 3A4, reduces serum concentrations of total bile acids only in individuals with liver disease and elevated serum bile acid levels. METHODS: We studied the effect of rifampin on concentrations of surrogate serum markers of cholesterol and bile acid synthesis as well as of cholesterol absorption in 10 male subjects before and after administration of rifampin (600 mg/day) for 6 days. Cholesterol and its precursors were analyzed by gas-liquid chromatography (GLC), bile acid intermediates and individual bile acids by isotope-dilution methods using GLC-mass spectrometry (MS) or by high-performance liquid chromatography (HPLC). RESULTS: Treatment with rifampin resulted in a 70% increase (p = 0.008) of the serum concentration of the bile acid precursor 7alpha-hydroxy-4-cholesten-3-one, which is a marker for bile acid production. Serum total cholesterol was not altered, however, treatment with rifampin elevated the ratio of lathosterol to cholesterol, an indicator of cholesterol synthesis, by 23% (p = 0.037). Interestingly, serum concentration of total bile acids decreased slightly by 29% (p = 0.022), mainly due to a lowering of the secondary bile acid, deoxycholic acid (-60%; p = 0.005). CONCLUSION: A 6-day treatment with rifampin induces a reduction of deoxycholic serum concentrations in healthy men associated with a moderate increase of serum markers of bile acid and endogenous cholesterol synthesis.