In Situ, Quantitative Assessment of Multifunctional Nanoscale Drug Delivery Systems in Human Serum.

The large volume and diversified nanomedicine market, undergoing a rapid growth, relies not only on the creation and applicative exploration of nanocarrier-based medicines showing significant potential, but in particular, demands a quantitative assessment of their physicochemical properties. In this study, we demonstrate the in situ assessment of multifunctional biodegradable nanoparticle (NP) entries as core components of nanoscale drug delivery systems (NDDSs) by making use of analytical ultracentrifugation (AUC). We determine and elucidate the following characteristics of NPs in NDDSs: NP density and size, targeting dye functionality, encapsulated and free drug, surfactant, and also NP drug release dynamics, quantitatively interconnected to NP degradation. In concept, we demonstrate this by multidetection AUC experiments at variable speed and time profiles. We could verify the quantitative and accurate nature of AUC for assessment of NDDSs, that is, also future nanomedicines. This concerns modeled and real life solution application formats such as cell culture media and human serum.

High variability of TB, HIV, hepatitis C treatment and opioid substitution therapy among prisoners in Germany.

BACKGROUND: In Germany, medical care of prisoners is completely separated from extramural health care. The extent and quality of medical care among prisoners in Germany are therefore largely unknown. We performed a secondary data analysis of pharmacy sales data for tuberculosis (TB), HIV, hepatitis C (HCV) and opioid substitution treatment (OST) delivered to prisons in 11 federal states (FS) in Germany between 01/2012 and 03/2013. The aims of this study were to assess (i) the treatment availability for the selected diseases and OST in German prisons, (ii) the proportion of prisoners treated per FS and overall for TB, HIV, HCV and OST during the study period. METHODS: Substances unique to or typically used for the treatment of each disease were defined as marker substances with defined daily doses (DDD). For each marker substance we assessed the cumulative number of DDD, the average daily number of DDD (DDDd) and average treatment prevalence per day in percent (adTP). Accordingly, the DDDd represents one person treated per day and the adTP means the proportion of prisoners treated per day. We compared the adTP of the diseases with previously measured prevalences. RESULTS: We obtained data from pharmacies supplying prisons in 11 of 16 German FS. Of the included prisons, 41% were supplied with medicines for TB, 71% for HIV and 58% for HCV and OST. Twice as many delivered marker substances for TB were indicated for the continuation phase and chemoprevention than the intensive phase. The HIV adTP ranged from 0.06% to 0.94%, HCV adTP ranged from 0.03% to 0.59% and OST adTP ranged from 0% to 7.90%. The overall adTP for the respective treatment was 0.39% for HIV, 0.12% for HCV and 2.18% for OST. CONCLUSIONS: According to our findings treatment rates for TB were consistent with the expected TB prevalence, at least in Berlin. HIV treatment seems to be offered to an adequate proportion of estimated infected prisoners. In contrast, the HCV treatment prevalence was low. High variation among FS in provision of all treatments, particularly of OST, point to inconsistent treatment practices, although nationwide extramural treatment guidelines for Germany exist.

Early citalopram treatment increases mortality due to left ventricular rupture in mice after myocardial infarction.

AIMS: Both anxiety and depression are common and independent outcome predictors in patients after myocardial infarction (MI). However, it is unclear whether and how anti-depressants influence remodeling after MI. Thus, we studied cardiac remodeling in mice after experimental MI under treatment with citalopram, a selective serotonin reuptake inhibitor widely used as antidepressant. METHODS AND RESULTS: Treatment with citalopram versus saline was applied via osmotic pump after coronary artery ligation. Two different groups were studied: early treatment during the healing phase (starting immediately after surgery), or late treatment in the remodeling phase (starting 7days after surgery). Late treatment did not change mortality or left ventricular remodeling after MI over the period of 6weeks. However, in the early treatment group mortality was increased in citalopram-treated mice predominantly due to left ventricle rupture without differences in infarct size. Remodeling 4weeks after MI was not altered by the treatment. Neither infiltration of inflammatory cells, as determined by FACS analysis of myocardial tissue, nor mRNA-expression of inflammatory cytokines changed 3days after MI in the early treatment group. However, extracellular matrix functioning was altered: There was a significant increase of MMP13 in citalopram treated animals after MI. Pretreatment with the MMP inhibitor PD 166793 prevented left ventricular ruptures and demonstrated a tendency to improved survival after citalopram treatment. CONCLUSIONS: Treatment with antidepressant citalopram in the acute but not in the late phase after MI significantly increased mortality in mice by disturbing early healing. Pharmacological MMP inhibition partially reversed the deleterious effects of citalopram.

Pathogen-specific DNA enrichment does not increase sensitivity of PCR for diagnosis of invasive aspergillosis in neutropenic patients.

PCR assays designed for the diagnosis of invasive aspergillosis (IA) in high-risk patients have to detect minute amounts of target DNA to reach sufficient analytical sensitivity to be of clinical use. This prospective study assessed the use of a novel strategy for selective pathogen DNA enrichment for enhancing the performance of diagnostic PCR in a direct comparison with a highly sensitive in-house quantitative PCR (qPCR) assay and the galactomannan enzyme-linked immunosorbent assay (ELISA). Surprisingly, and in contrast to experience with other patient groups, the novel protocol for selective pathogen DNA enrichment did not enhance but instead significantly impaired sensitivity. This could be explained by the small amounts of host DNA in the specimens, which were derived mostly from severely neutropenic patients. In the qPCR assay, positive samples required an average of 43.5 amplification cycles (range, 39.2 to 50) for detection in the in-house PCR. Repetitive testing of selected samples showed test positivity to be variable, most likely due to the small amounts of target DNA. Despite this, the in-house protocol proved helpful in the diagnosis of IA, detecting 2 out of 3 patients with probable IA and 10 out of 19 patients with possible IA. Our results underline the necessity for diagnostic PCR protocols that help diagnose IA to be highly sensitive and show that selective pathogen DNA enrichment using affinity purification may not be useful in severely neutropenic patients.

Prevalence of Opioid Dependence and Opioid Agonist Treatment in the Berlin Custodial Setting: A Cross-Sectional Study.

BACKGROUND: Among people living in detention, substance use is highly prevalent, including opioid dependence. Opioid agonist treatment (OAT) has been established as an evidence-based, first-line treatment for opioid dependence. Despite high prevalence of opioid dependence, conclusive data regarding its prevalence and the OAT practice in German prisons is scarce; rather, the existing data widely diverges concerning the rates of people in detention receiving OAT. MATERIALS AND METHODS: We conducted a cross-sectional survey of all detention facilities in Berlin. On the date of data collection, a full census of the routine records was completed based on the medical documentation system. For each opioid dependent individual, we extracted sociodemographic data (i.e., age, sex, and non-/German nationality, whether people experienced language-related communication barriers), information about OAT, comorbidities (HIV, hepatitis C, schizophrenia), and the detention center, as well as the anticipated imprisonment duration and sentence type. The data was first analyzed descriptively and secondly in an evaluative-analytical manner by analyzing factors that influence the access to OAT of people living in detention. RESULTS: Among the 4,038 people in detention in the Berlin custodial setting under investigation, we identified a 16% prevalence of opioid dependence. Of the opioid-dependent individuals, 42% received OAT; 31% were treated with methadone, 55% were treated with levomethadone, and 14% were treated with buprenorphine. Access to OAT seemed mainly dependent upon initial receipt of OAT at the time of imprisonment, detention duration, the prisons in which individuals were detained, German nationality, and sex. The overall prevalence of HIV was 4-8%, hepatitis C was 31-42%, and schizophrenia was 5%. CONCLUSIONS: The prevalence of opioid dependence and access to OAT remains a major health issue in the custodial setting. OAT implementation must be especially intensified among male, non-German, opioid-dependent individuals with a short detention period. Treatment itself must be diversified regarding the substances used for OAT, and institutional treatment differences suggest the need for a consistent treatment approach and the standardized implementation of treatment guidelines within local prison's standard operating procedures. Testing for infectious diseases should be intensified among opioid-dependent people living in detention to address scarcely known infection statuses and high infection rates.

Formulation of Liver-Specific PLGA-DY-635 Nanoparticles Loaded with the Protein Kinase C Inhibitor Bisindolylmaleimide I.

Bisindolylmaleimide I (BIM-I) is a competitive pan protein kinase C inhibitor with anti-inflammatory and anti-metastatic properties, suggested to treat inflammatory diseases and various cancer entities. However, despite its therapeutic potential, BIM-I has two major drawbacks, i.e., it has a poor water solubility, and it binds the human ether-à-go-go-related gene (hERG) ion channels, potentially causing deadly arrhythmias. In this case, a targeted delivery of BIM-I is imperative to minimize peripheral side effects. To circumvent these drawbacks BIM-I was encapsulated into nanoparticles prepared from poly(lactic-co-glycolic acid) (PLGA) functionalized by the near-infrared dye DY-635. DY-635 served as an active targeting moiety since it selectively binds the OATP1B1 and OATP1B3 transporters that are highly expressed in liver and cancer cells. PLGA-DY-635 (BIM-I) nanoparticles were produced by nanoprecipitation and characterized using dynamic light scattering, analytical ultracentrifugation, and cryogenic transmission electron microscopy. Particle sizes were found to be in the range of 20 to 70 nm, while a difference in sizes between the drug-loaded and unloaded particles was observed by all analytical techniques. In vitro studies demonstrated that PLGA-DY-635 (BIM-I) NPs prevent the PKC activation efficiently, proving the efficacy of the inhibitor after its encapsulation, and suggesting that BIM-I is released from the PLGA-NPs. Ultimately, our results present a feasible formulation strategy that improved the cytotoxicity profile of BIM-I and showed a high cellular uptake in the liver as demonstrated in vivo by intravital microscopy investigations.

Polymethine Dye-Functionalized Nanoparticles for Targeting CML Stem Cells.

In chronic myelogenous leukemia (CML), treatment with tyrosine kinase inhibitors (TKI) is unable to eradicate leukemic stem cells (LSC). Polymethine dye-functionalized nanoparticles can be internalized by specific cell types using transmembrane carrier proteins. In this study we investigated the uptake behavior of various polymethine dyes on leukemia cell lines and searched for carrier proteins that guide dye transport using RNA interference. The results show that the uptake of DY-635 is dependent on organic anion transport protein 1B3 (OATP1B3) in CML cells and immature myeloid precursor cells of CML patients. In contrast to nonspecific poly(lactide-co-glycolic acid) (PLGA) nanoparticle constructs, DY-635-functionalization of nanoparticles led to an uptake in CML cells. Investigation of these nanoparticles on bone marrow of CML patients showed a preferred uptake in LSC. The transcription of OATP1B3 is known to be induced under hypoxic conditions via the hypoxia-inducing factor 1 alpha (HIF1α), thus also in the stem cells niche. Since these cells have the potential to repopulate the bone marrow after CML treatment discontinuation, eliminating them by means of drug-loaded DY-635-functionalized PLGA nanoparticles deployed as a selective delivery system to LSC is highly relevant to the ongoing search for curative treatment options for CML patients.

Truncated human cytidylate-phosphate-deoxyguanylate-binding protein for improved nucleic acid amplification technique-based detection of bacterial species in human samples.

A trunk of human cytidylate-phosphate-deoxyguanylate-binding protein/CXXC finger protein 1 (CFP1), immobilized onto an aminohexyl-Sepharose column, can be used as a preanalytical tool for the selective enrichment of bacterial DNA from mixed solutions with high amounts of human background DNA for nucleic acid amplification technique-based detection of pathogens. The transcriptional activator protein exhibits a high affinity for nonmethylated CpG dinucleotide motifs, which are differentially distributed in prokaryotic and higher eukaryotic genomes. The feasibility of the affinity chromatography (AC) step was tested with DNA from severely septic patients. AC using 16S rRNA gene primers substantially increased PCR sensitivity. Approximately 90% of eukaryotic DNA was removed, which significantly increased the signal-to-noise ratio. Threshold cycle values revealed that sensitivity was elevated at least 10-fold. The change in the ratio of bacterial DNA to human DNA increased from 26% to 74% the likelihood of culture-independent PCR-based identification of bacterial presence. Compared to the results seen with blood culture (which is the clinical gold standard for systemic infections, exhibiting 28% positives), the combination of AC and PCR achieves a significant increase in sensitivity and contributes to shortening the time to results for the initiation of guided antibiotic therapy.

Identification of bacterial DNA in neutrocytic and non-neutrocytic cirrhotic ascites by means of a multiplex polymerase chain reaction.

BACKGROUND: Even though bacterial cultures of ascitic fluid are negative in up to 65% of the cases of spontaneous bacterial peritonitis (SBP); bacterial DNA (bactDNA) has been frequently detected in episodes of SBP as well as in culture-negative non-neutrocytic ascites. AIMS: To evaluate multiplex polymerase chain reaction (PCR) for pathogen identification in SBP and to determine the prevalence of ascitic bactDNA and its prognostic relevance in hospitalized patients with liver cirrhosis. METHODS: Ascitic fluid from 68 consecutive patients who underwent diagnostic paracentesis was analysed for polymorphonuclear leucocyte (PMN) count, bacterial culture and bactDNA. BactDNA was identified by gel analysis after multiplex PCR of selectively enriched prokaryotic nucleic acids. Correlations of bactDNA status with PMN count, bacterial culture result and 3-month mortality were determined for neutrocytic and for non-neutrocytic ascites. RESULTS: 11/68 patients presented with an elevated ascitic PMN count. BactDNA was detected in 5/5 culture-positive neutrocytic samples, in 1/6 culture-negative neutrocytic samples and in 8/56 culture-negative non-neutrocytic samples. Three-month mortality did not differ with respect to ascitic bactDNA status (7/14 vs. 14/47, P=0.162). 3-month mortality was increased in the presence of ascitic bactDNA for patients older than 65 years (4/5 vs. 4/14, P=0.046) and for patients with a model for end-stage liver disease score >15 (7/10 vs. 9/30, P=0.025). CONCLUSIONS: Identification of ascitic bactDNA is an appropriate alternative to bacterial ascite culture for pathogen identification in patients at risk for SBP. Its prognostic relevance as a proposed marker of bacterial translocation for certain risk groups has to be further evaluated.

Medicine in the Penal System.

BACKGROUND: Infectious diseases, substance dependencies, and dental diseases are the most important health problems affecting incarcerated persons. In Germany, for example, prisoners are 48 to 69 times more likely to be infected with the hepatitis C virus (HCV) than the general population, and 7 to 12 times more likely to be infected with the human immunodeficiency virus (HIV). The prevalence of mental illnesses is also markedly higher in the incarcerated than in the general population. METHODS: This review is based on pertinent publications retrieved by a selective search in two databases (PubMed and Google Scholar) for any of the terms "health care," "primary health care," "mental health care"; "infectious disease," "opioid maintenance treatment," and "severe mental disorder" in conjunction with "prison," "jail," "detention," and "incarceration." RESULTS: Among prisoners in German prisons, approximately 20% consume heroin, 20-50% suffer from alcohol dependency and abuse, and 70-85% smoke. The prevalence of tuberculosis in German prisons in 2002 was 0.1%. The provision of needles to incarcerated persons has a preventive effect on infection with hepatitis C, hepatitis B, and HIV, yet programs of this type have been discontinued in most penal facilities. In a systematic review, psychotic disorders were found in 3.6% (95% confidence interval [CI]: [3.1; 4.2]) of male inmates and 3.9% [95% CI: 2.7; 5.0] of female inmates. 25% of incarcerated persons suffer from attention-deficit-hyperac- tivity disorder. Persons recently released from prison have an above average mortality, largely due to drug intoxication. CONCLUSION: An analysis of medical prescribing data reveals deficiencies in the provision of HCV treatment to all affected persons and in the provision of substitution treatment to persons with opiate dependency. In view of the known risks associated with imprisonment, greater emphasis should be placed on the provision of treatment for infectious diseases, substance dependencies, and mental illness, both in prison and in outpatient care after release.

Clearance of low levels of HCV viremia in the absence of a strong adaptive immune response.

Spontaneous clearance of hepatitis C virus (HCV) has frequently been associated with the presence of HCV-specific cellular immunity. However, there had been also reports in chimpanzees demonstrating clearance of HCV-viremia in the absence of significant levels of detectable HCV-specific cellular immune responses. We here report seven asymptomatic acute hepatitis C cases with peak HCV-RNA levels between 300 and 100,000 copies/ml who all cleared HCV-RNA spontaneously. Patients were identified by a systematic screening of 1176 consecutive new incoming offenders in a German young offender institution. Four of the seven patients never developed anti-HCV antibodies and had normal ALT levels throughout follow-up. Transient weak HCV-specific CD4+ T cell responses were detectable in five individuals which did not differ in strength and breadth from age- and sex-matched patients with chronic hepatitis C and long-term recovered patients. In contrast, HCV-specific MHC-class-I-tetramer-positive cells were found in 3 of 4 HLA-A2-positive patients. Thus, these cases highlight that clearance of low levels of HCV viremia is possible in the absence of a strong adaptive immune response which might explain the low seroconversion rate after occupational exposure to HCV.

Chlamydia pneumoniae infection of aortic smooth muscle cells reduces platelet-derived growth factor receptor-beta expression.

Chlamydia pneumoniae infection may play a role in the pathogenesis of atherosclerosis. In this study, an oligonucleotide microarray was utilized to examine the transcriptional response of human aortic smooth muscle cells (AoSMC) to C. pneumoniae infection. Alteration of mRNA expression in 71 out of 780 genes was detected at 24 h after infection. Among the down-regulated genes, platelet-derived growth factor receptor-beta (PDGFR-beta) was identified as a target for further analysis because the PDGF system is involved in the fibroproliferative response of SMC in atherogenesis. Reverse transcriptase PCR analysis demonstrated that C. pneumoniae inhibits the up-regulation of PDGFR-beta mRNA occurring in AoSMC after mock infection. PDGFR-beta protein synthesis was examined by immunoblotting and fluorescence-activated cell sorting. Compared with mock-infected cells, the amount of receptor protein was reduced at 24, 48, and 72 h after infection. Diminished PDGFR-beta synthesis in infected cultures was accompanied by the suppression of AoSMC growth following PDGF-BB stimulation. The interference of C. pneumoniae with PDGFR-beta expression may result in decreased SMC proliferation in atherosclerotic plaques, thereby affecting the development and stability of advanced lesions.

Mission ImPOxable - or the unknown utilization of non-toxic poly(2-oxazoline)s as cryoprotectants and surfactants at the same time.

Polymer based nanoparticles offer great opportunities for diverse applications, i.e. their drug delivery potential is promising. However, their major drawback is identified in preparation via the nanoemulsion technique, which is needed for the encapsulation of hydrophilic drugs and whereby the utilization of surfactants, e.g. poly(vinyl alcohol) (PVA), is mandatory. Furthermore, the preparation of nanoparticles is critical due to the need of lyophilization for storage. For this reason it is common to use cryoprotectants, which are usually sugar based. In the current study, we present the use of non-toxic, water-soluble poly(2-oxazoline)s (P(Ox)s) in terms of polymeric nanoparticle stabilizers for preparation, purification, and lyophilization. The nanoparticles were characterized via dynamic light scattering (DLS) and cryo-transmission electron microscopy (cryoTEM). The use of hydrophilic P(Ox)s with a degree of polymerization of about 60 yielded stable nanoparticles. For the preparation via nanoemulsion a PDI below 0.2 could be obtained after adjustment of the surfactant concentration. All nanoparticles were in the size range of 100 to 200 nm according to DLS. Furthermore, the addition of P(Ox) was beneficial during particle purification via centrifugation and filtration as well as lyophilization, yielding nanoparticles with a PDI below 0.3 as determined via DLS and confirmed via cryoTEM measurements. Cytotoxicity, hemolysis and erythrocyte aggregation measurements of these P(Ox)s did not show any harmful effect on the treated cells.

Experimental heart failure causes depression-like behavior together with differential regulation of inflammatory and structural genes in the brain.

BACKGROUND: Depression and anxiety are common and independent outcome predictors in patients with chronic heart failure (CHF). However, it is unclear whether CHF causes depression. Thus, we investigated whether mice develop anxiety- and depression-like behavior after induction of ischemic CHF by myocardial infarction (MI). METHODS AND RESULTS: In order to assess depression-like behavior, anhedonia was investigated by repeatedly testing sucrose preference for 8 weeks after coronary artery ligation or sham operation. Mice with large MI and increased left ventricular dimensions on echocardiography (termed CHF mice) showed reduced preference for sucrose, indicating depression-like behavior. 6 weeks after MI, mice were tested for exploratory activity, anxiety-like behavior and cognitive function using the elevated plus maze (EPM), light-dark box (LDB), open field (OF), and object recognition (OR) tests. In the EPM and OF, CHF mice exhibited diminished exploratory behavior and motivation despite similar movement capability. In the OR, CHF mice had reduced preference for novelty and impaired short-term memory. On histology, CHF mice had unaltered overall cerebral morphology. However, analysis of gene expression by RNA-sequencing in prefrontal cortical, hippocampal, and left ventricular tissue revealed changes in genes related to inflammation and cofactors of neuronal signal transduction in CHF mice, with Nr4a1 being dysregulated both in prefrontal cortex and myocardium after MI. CONCLUSIONS: After induction of ischemic CHF, mice exhibited anhedonic behavior, decreased exploratory activity and interest in novelty, and cognitive impairment. Thus, ischemic CHF leads to distinct behavioral changes in mice analogous to symptoms observed in humans with CHF and comorbid depression.

Psychiatric care in the German prison system.

PURPOSE: The purpose of this paper is to describe the nature of medical care within the German penal system. German prison services provide health care for all inmates, including psychiatric care. The reached level of equivalence of care and ethical problems and resource limitations are discussed and the way of legislation in this field since 2006 reform on federal law is described. DESIGN/METHODOLOGY/APPROACH: The article summarizes basic data on German prison health care for mentally ill inmates. The legislation process and factors of influence are pointed out. A description of how psychiatric care is organized in German prisons follows. It focuses on the actual legal situation including European standards of prison health care and prevention of torture, psychiatric care in German prisons themselves, self harm and addiction. Associated problems such as blood born diseases and tuberculosis are included. The interactions between prison staff and health care personal and ethic aspects are discussed. FINDINGS: The legislation process is still going on and there is still a chance to improve psychiatric care. Mental health problems are the major challenge for prison health care. Factors such as special problems of migrants, shortage of professionals and pure statistic data are considered. ORIGINALITY/VALUE: The paper provides a general overview on psychiatric services in prison and names weak points and strengths of the system.

Evaluation of a polymerase chain reaction assay for pathogen detection in septic patients under routine condition: an observational study.

BACKGROUND: Treatment of septic shock relies on appropriate antimicrobial therapy. Current culture based methods deliver final results after days, which may delay potentially lifesaving adjustments in antimicrobial therapy. This study was undertaken to compare PCR with blood culture results under routine conditions regarding 1. impact on antimicrobial therapy, and 2. time to result, in patients with presumed sepsis. METHODOLOGY/PRINCIPAL FINDINGS: This was an observational study in a 50 beds ICU of a university hospital. In 245 patients with suspected sepsis, 311 concomitant blood cultures and blood for multiplex PCR (VYOO(®)) were obtained. 45 of 311 blood cultures (14.5%) and 94 of 311 PCRs (30.1%) were positive. However, blood culture or microbiological sampling from the presumed site of infection rarely confirmed PCR results and vice versa. Median time to positivity and interquartile range were 24.2 (18.0, 27.5) hours for the PCR and 68 (52.2, 88.5) hours for BC (p<0.01). PCR median time to result was dependent on technician availability (53.5 hours on Saturdays, 7.2 hours under optimal logistic conditions). PCR results showed good correlation with procalcitonin (p<0.001). In 34% of patients with positive PCRs antimicrobial therapy was considered inadequate according to assessment of clinical arbitrators including 5 patients with vancomycin-resistant enterococci (VRE), 3 cases with multiresistant staphylococci, and 4 patients with fungi. CONCLUSIONS: The results of this observational study support the hypothesis that PCR results are available faster, are more frequently positive, and may result in earlier adjustment of antimicrobial therapy. However, shorter time to result can only be fully exploited when the laboratory is adequately staffed for a 24 hour/7 day service, or when point of care/automated assay systems become available.

Human cytomegalovirus induces MMP-1 and MMP-3 expression in aortic smooth muscle cells.

Human cytomegalovirus (HCMV) infection may be involved in the pathogenesis of atherosclerosis by modulating functions of smooth muscle cells (SMC). In this study, we performed an oligonucleotide microarray screening of 780 inflammation-associated genes in HCMV-infected aortic SMC (AoSMC). The expression of 31 genes was stimulated and 24 genes were down-regulated following infection with HCMV strain DC-134. Following infection with HCMV strain AD-169 infection, we found 24 genes to be stimulated and 32 genes to be down-regulated. Among these were primarily genes encoding for CC and CXC chemokines, adhesion molecules, and tumor necrosis factor (TNF) receptor superfamily members, apoptosis-related factors, signal transduction molecules and transcription regulators. The up-regulated genes included matrix metalloproteinase (MMP)-1 and MMP-3 in HCMV infected cells. Using RT-PCR and enzyme immunoassay we found stimulated expression of MMP-1 (3.2-fold expression) and MMP-3 (334-fold expression) in HCMV strain DC-134-infected AoSMC at 72 h following infection.The findings of our study suggest that HCMV infection of AoSMC cause an activation of atherosclerosis-relevant factors in SMC. The increased expression of MMPs which have been shown to be involved in atherosclerotic plaque rupture and myocardial infarction is in agreement with the hypothesis that this pathogen might contribute to plaque inflammation in atherosclerotic disease.

Effects of a multidisciplinary inpatient intervention on body composition, aerobic fitness, and quality of life in severely obese girls and boys.

PURPOSE: The last few decades have seen a major increase in the prevalence of juvenile obesity. Inpatient treatment programs are used mainly in children with severe obesity and related comorbidities. The aim of this study was to evaluate the gender differences of an 8-week multidisciplinary inpatient program on body weight, body composition, aerobic fitness, and quality of life of severely obese children and adolescents. METHODS: Body weight was measured daily, and body composition, aerobic fitness, and quality of life were measured at the beginning and the end of an 8-week multidisciplinary inpatient program in 130 severely obese patients (52 girls, 78 boys), median (25th, 75th percentile) age of 13.8 (12.1, 15.0) years, median body weight of 89.4 kg (77.1, 100.1), and a body mass index of 33.4 (30.1, 36.6) kg/m(2), which is well above the 98th percentile. The inpatient program was based on a multidisciplinary treatment and education program that focused on daily physical activity, a 1200-1600 kcal/day balanced nutrition regimen, and a behavior modification therapy. RESULTS: All results are expressed as medians (25th, 75th percentiles). At the end of the program all patients had lost a significant amount of body weight: 12.7 kg (10.8, 16.6), p < .001, girls 11.6 kg (9.7, 13.2), boys 13.7 kg (11.7, 17.3), p < .001, absolute body fat 8.0 kg (6.8, 10.0) p < 001, girls 7.0 kg (5.7, 8.1), boys 9.4 kg (7.6, 11.0) p < .001, % body fat per kg body weight: 4.9% (3.2, 6.6) p < .001, girls 3.7% (2.7, 4.9), boys 5.7% (4.0, 7.5) p < .001, and absolute fat free (or lean body) mass: 1.8 kg (0.64, 3.0) p < .001, girls 1.8 kg (0.87, 3.2), boys 1.7 kg (0.50, 2.9) p = .43. In addition, all measurements of aerobic fitness: VO(2)peak (mL/min.kg) and peak mechanical power (watts and watt/kg) and of quality of life increased significantly (p < .001, p < .001, p < .004 to p < .001). CONCLUSION: A multidisciplinary inpatient treatment program including moderate calorie restriction, daily physical activity, and behavior modification induced a major weight loss, a decrease in body fat, and an increse in aerobic fitness as well as the quality fo life of severely obese children and adolescents. Weight loss and the decrease in body fat (absolute and percent) were significantly more pronounced in boys than girls.

High rate of spontaneous clearance of acute hepatitis C virus genotype 3 infection.

Treating acute hepatitis C with interferon alpha prevents chronicity in nearly all cases when therapy is initiated within 3 months after infection. However, 15-50% of untreated patients may clear the hepatitis C virus (HCV) spontaneously. Therefore, factors are needed to identify patients prior to therapy who have a higher or lower risk for developing a chronic course to avoid unnecessary treatment. The role of the HCV genotype for spontaneous recovery from acute hepatitis C has been discussed controversially. In the year 2002, all 1,176 new incoming prisoners in a Northern German prison for young men (age 16-24) were screened for anti-HCV antibodies and 92 tested positive. Ninety eight percent of these reported i.v.-drug abuse for a median of 32 months prior to imprisonment. HCV-RNA negative individuals (21%) were serotyped and HCV-RNA positive patients were genotyped. The prevalence of HCV genotype 3 was significantly higher among individuals who had cleared HCV spontaneously as compared to chronically infected patients (86% vs. 38%; P = 0.002). Ninety three percent of individuals exposed to HCV genotype 1 but only 63% of individuals exposed to genotype 3 experienced a chronic course of the infection (P = 0.006). Thus, acute infection in young Caucasian men with HCV genotype 3 leads more often to spontaneous clearance than infection with HCV genotype-1. Considering also the high chance of successful treatment of chronic HCV genotype 3 infection with pegylated-interferon in combination with ribavirin, we suggest not to treat acute hepatitis C genotype 3 infection early but rather to wait at least 3 months after the onset of symptoms when chronicity becomes likely.