Epstein-Barr Virus-Induced Genes and Endogenous Retroviruses in Immortalized B Cells from Patients with Multiple Sclerosis.

The immune pathogenesis of multiple sclerosis (MS) is thought to be triggered by environmental factors in individuals with an unfavorable genetic predisposition. Epstein-Barr virus (EBV) infection is a major risk factor for subsequent development of MS. Human endogenous retroviruses (HERVs) can be activated by EBV, and might be a missing link between an initial EBV infection and the later onset of MS. In this study, we investigated differential gene expression patterns in EBV-immortalized lymphoblastoid B cell lines (LCL) from MS-affected individuals (MSLCL) and controls by using RNAseq and qRT-PCR. RNAseq data from LCL mapped to the human genome and a virtual virus metagenome were used to identify possible biomarkers for MS or disease-relevant risk factors, e.g., the relapse rate. We observed that lytic EBNA-1 transcripts seemed to be negatively correlated with age leading to an increased expression in LCL from younger PBMC donors. Further, HERV-K (HML-2) GAG was increased upon EBV-triggered immortalization. Besides the well-known transactivation of HERV-K18, our results suggest that another six HERV loci are up-regulated upon stimulation with EBV. We identified differentially expressed genes in MSLCL, e.g., several HERV-K loci, ERVMER61-1 and ERV3-1, as well as genes associated with relapses. In summary, EBV induces genes and HERV in LCL that might be suitable as biomarkers for MS or the relapse risk.

Headache in Multiple Sclerosis - Pharmacological Aspects.

For decades, the headache was not considered a typical symptom of multiple sclerosis (MS) and was construed as a "red flag" for important differential diagnoses such as cerebral vasculitis. Meanwhile, several studies have demonstrated an increased prevalence of headaches in MS compared to the general population. This is due to the heterogeneity of headache genesis with frequent occurrence of both primary and secondary headaches in MS. On the one hand, MS and migraine are often comorbid. On the other hand, secondary headaches frequently occur, especially in the course of MS relapses. These are often migraine-like headaches caused by inflammation, which can improve as a result of MS-specific therapy. Headaches are particularly common in the early stages of chronic inflammatory CNS disease, where inflammatory activity is the greatest. In addition, headaches can also occur as a side effect of disease-modifying drugs (DMDs). Headache can occur with most DMDs and is most frequently described with interferon-beta therapy. The aim of this work is to present the prevalence of headaches and describe the heterogeneity of possible causes of headaches in MS. In addition, important therapeutic aspects in the treatment of MS patients, in general, will be presented as well as different approaches to the treatment of headaches in MS depending on the etiological classification.

Investigation of Endogenous Retrovirus Sequences in the Neighborhood of Genes Up-regulated in a Neuroblastoma Model after Treatment with Hypoxia-Mimetic Cobalt Chloride.

Human endogenous retroviruses (ERVs) have been found to be associated with different diseases, e.g., multiple sclerosis (MS). Most human ERVs integrated in our genome are not competent to replicate and these sequences are presumably silent. However, transcription of human ERVs can be reactivated, e.g., by hypoxia. Interestingly, MS has been linked to hypoxia since decades. As some patterns of demyelination are similar to white matter ischemia, hypoxic damage is discussed. Therefore, we are interested in the association between hypoxia and ERVs. As a model, we used human SH-SY5Y neuroblastoma cells after treatment with the hypoxia-mimetic cobalt chloride and analyzed differences in the gene expression profiles in comparison to untreated cells. The vicinity of up-regulated genes was scanned for endogenous retrovirus-derived sequences. Five genes were found to be strongly up-regulated in SH-SY5Y cells after treatment with cobalt chloride: clusterin, glutathione peroxidase 3, insulin-like growth factor 2, solute carrier family 7 member 11, and neural precursor cell expressed developmentally down-regulated protein 9. In the vicinity of these genes we identified large (>1,000 bp) open reading frames (ORFs). Most of these ORFs showed only low similarities to proteins from retro-transcribing viruses. However, we found very high similarity between retrovirus envelope sequences and a sequence in the vicinity of neural precursor cell expressed developmentally down-regulated protein 9. This sequence encodes the human endogenous retrovirus group FRD member 1, the encoded protein product is called syncytin 2. Transfection of syncytin 2 into the well-characterized Ewing sarcoma cell line A673 was not able to modulate the low immunostimulatory activity of this cell line. Future research is needed to determine whether the identified genes and the human endogenous retrovirus group FRD member 1 might play a role in the etiology of MS.

Copper(ii) mixed-ligand polypyridyl complexes with doxycycline - structures and biological evaluation.

Mixed-ligand Cu(ii) complexes of the type [Cu(doxycycline)(L)(H2O)2](NO3)2, where doxycycline = [4-(dimethylamino)-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide] and L = 2,2'-bipyridine (bpy, 1), 1,10-phenanthroline (phen, 2), dipyrido[3,2-d:2',3'-f]quinoxaline (dpq, 3) and dipyrido[3,2-a:2',3'-c]phenazine (dppz, 4) have been synthesised and characterised by structural, analytical, and spectral methods. The single-crystal X-ray structures of 1 and 2 exhibited two different geometries, distorted square-pyramidal and octahedral respectively as well as different coordination modes of doxycycline. Complexes 2-4 exhibit prominent plasmid DNA cleavage at significantly low concentrations probably by an oxidative mechanism. Matrix Metalloproteinase (MMP-2) inhibition studies revealed that all complexes inhibit MMP-2 similar to doxycycline which is a well-known MMP inhibitor with 3 being the most potent. IC50 values of doxycycline and 1-4 against MCF-7 (human breast cancer) and HeLa cell lines were almost equal in which 3 showed the highest efficiency (IC50 = 0.46 ± 0.05 µM), being consistent with its increased MMP inhibition potency. The antimalarial activities of these complexes against the chloroquine-sensitive Plasmodium falciparum NF54 and chloroquine-resistant Plasmodium falciparum Dd2 strains reveal that complex 3 exhibited a higher activity than artesunate drug against the chloroquine-resistant Dd2 strain.

Crystal structures of 1-bromo-3,5-bis-(4,4-dimethyl-1,3-oxazolin-2-yl)benzene 0.15-hydrate and 3,5-bis-(4,4-dimethyl-1,3-oxazolin-2-yl)-1-iodo-benzene.

The bromo and iodo derivatives of a meta-bis-(1,3-oxazolin-2-yl)-substituted benzene, C16H19BrN2O2·0.15H2O (1) and C16H19IN2O2 (2), have been prepared and studied in terms of their mol-ecular and crystal structures. While the former crystallizes as a sub-hydrate, with 0.15 formula units of water and shows an almost all-planar arrangement of the three ring systems, the latter crystallizes solvate-free with the flanking heterocycles twisted considerably with respect to the central arene. Non-covalent contacts include parallel-displaced π-π inter-actions and (non-classical) hydrogen bonding for both (1) and (2), as well as relatively short I⋯N contacts for (2).

The Patella Pro study - effect of a knee brace on patellofemoral pain syndrome: design of a randomized clinical trial (DRKS-ID:DRKS00003291).

BACKGROUND: Patellofemoral pain syndrome (PFPS) is a frequent cause of anterior knee pain predominantly affecting young female patients who do not have significant chondral damage. Development of PFPS is probably multifactorial, involving various knee, hip, and foot kinematic factors. Biomechanical studies have described patellar maltracking and dynamic valgus (functional malalignment) in patients with patellofemoral pain syndrome. The literature provides evidence for short-term use of nonsteroidal anti-inflammatory drugs; short-term medially directed taping; and exercise programs focusing on the lower extremity, hip, and trunk muscles. Evidence supporting the use of patellar braces is limited because previous studies have been low quality. The aim of this article is to publish the design of a prospective randomized trial that examines the outcomes of patients with PFPS after treatment with a new patellar brace (Patella Pro) that applies medially directed force on the patella. METHODS/DESIGN: For this multicenter trial, 156 patients (adolescents and young adults) with PFPS were recruited from orthopedic practices and orthopedic hospitals and randomly allocated to 3 months of supervised physiotherapy in combination with the Patella Pro brace or supervised physiotherapy alone. The primary outcome measures are pain (numerical analog scale); knee function (Kujala score and Knee Injury and Osteoarthritis Outcome Score); and self-reported perception of recovery at baseline, 6 weeks, 3 months, and 1 year. DISCUSSION: Only limited evidence for the use of a patellar brace for the treatment of PFPS exists in the literature. Disputable evidence for the use of orthoses for PFPS patients has been presented in one meta-analysis, in which only one of three studies found the effect of a medially directed patellar brace to be significant. Because of these low-quality studies, the authors concluded that this evidence should be regarded as limited, and we feel there is a need for further well-designed studies to evaluate the effect of patellar bracing on PFPS-related pain. The Patella Pro study is a prospective randomized trial in which supervised physiotherapy in combination with a patellar brace is compared with supervised physiotherapy alone. This trial started in April 2012 and finished in October 2013. TRIAL REGISTRATION: DRKS-ID:DRKS00003291, January 3rd, 2012.

Contrasting disease patterns in seropositive and seronegative neuromyelitis optica: A multicentre study of 175 patients.

BACKGROUND: The diagnostic and pathophysiological relevance of antibodies to aquaporin-4 (AQP4-Ab) in patients with neuromyelitis optica spectrum disorders (NMOSD) has been intensively studied. However, little is known so far about the clinical impact of AQP4-Ab seropositivity. OBJECTIVE: To analyse systematically the clinical and paraclinical features associated with NMO spectrum disorders in Caucasians in a stratified fashion according to the patients' AQP4-Ab serostatus. METHODS: Retrospective study of 175 Caucasian patients (AQP4-Ab positive in 78.3%). RESULTS: Seropositive patients were found to be predominantly female (p < 0.0003), to more often have signs of co-existing autoimmunity (p < 0.00001), and to experience more severe clinical attacks. A visual acuity of ≤ 0.1 during acute optic neuritis (ON) attacks was more frequent among seropositives (p < 0.002). Similarly, motor symptoms were more common in seropositive patients, the median Medical Research Council scale (MRC) grade worse, and MRC grades ≤ 2 more frequent, in particular if patients met the 2006 revised criteria (p < 0.005, p < 0.006 and p < 0.01, respectively), the total spinal cord lesion load was higher (p < 0.006), and lesions ≥ 6 vertebral segments as well as entire spinal cord involvement more frequent (p < 0.003 and p < 0.043). By contrast, bilateral ON at onset was more common in seronegatives (p < 0.007), as was simultaneous ON and myelitis (p < 0.001); accordingly, the time to diagnosis of NMO was shorter in the seronegative group (p < 0.029). The course of disease was more often monophasic in seronegatives (p < 0.008). Seropositives and seronegatives did not differ significantly with regard to age at onset, time to relapse, annualized relapse rates, outcome from relapse (complete, partial, no recovery), annualized EDSS increase, mortality rate, supratentorial brain lesions, brainstem lesions, history of carcinoma, frequency of preceding infections, oligoclonal bands, or CSF pleocytosis. Both the time to relapse and the time to diagnosis was longer if the disease started with ON (p < 0.002 and p < 0.013). Motor symptoms or tetraparesis at first myelitis and > 1 myelitis attacks in the first year were identified as possible predictors of a worse outcome. CONCLUSION: This study provides an overview of the clinical and paraclinical features of NMOSD in Caucasians and demonstrates a number of distinct disease characteristics in seropositive and seronegative patients.

An interpenetrated metal-organic framework and its gas storage behavior: simulation and experiment.

A new metal-organic framework, called UHM-6 (UHM: University of Hamburg Materials), based on the copper paddle wheel motif and a novel organosilicon linker, 4',4″-(dimethylsilanediyl)bis(biphenyl-3,5-dicarboxylic acid) (sbbip), has been synthesized and characterized with regard to its gas storage behavior up to 1 bar for hydrogen, methane, and carbon dioxide. The 2-fold interpenetrated microporous framework of UHM-6 is isoreticular to PMOF-3 (Inorg. Chem.2009, 48, 11507) and is composed of cuboctahedral cages of Cu(2) paddle wheels connected via nonlinear organosilicon units. The structure (SG I422, No. 97) is characterized by straight channels running along the [001] and [110] direction. UHM-6 reveals a specific surface area of S(BET) ~ 1200 m(2) g(-1) and a specific micropore volume of V(micropore) ~ 0.48 cm(3) g(-1). At 1 bar the activated form of UHM-6 shows a hydrogen uptake of 1.8 wt % (77 K), a methane uptake of 0.8 mmol g(-1) (293 K), and a carbon dioxide uptake of 3.3 mmol g(-1) (273 K). Accompanying theoretical grand-canonical Monte Carlo (GCMC) simulations show an overall good agreement with the experimental results. Furthermore, GCMC adsorption simulations for three binary equimolar mixtures (CH(4)/H(2), CO(2)/H(2), and CO(2)/CH(4)) were carried out (T = 298 K) to assess the potential for gas separation/purification applications.

A randomized study of the effectiveness of suprascapular nerve block in patient satisfaction and outcome after arthroscopic subacromial decompression.

PURPOSE: The purpose of this study was to evaluate the efficiency of the suprascapular nerve (SSN) block in pain reduction after arthroscopic subacromial decompression operations and its influence on patient satisfaction. Furthermore, we wanted to evaluate whether better perioperative pain management could positively influence postoperative shoulder function. METHODS: In this prospective, randomized, double-blinded clinical trial, 3 groups of patients--each with 15 participants--were treated with SSN block (10 mL of 1% ropivacaine), placebo, or a subacromial infiltration of local anesthesia (20 mL of 1% ropivacaine). Preoperative and postoperative pain was evaluated with a visual analog scale. Functional outcome was measured by the Constant-Murley score, and patient satisfaction was measured anecdotally by interview 2 days, 2 weeks, and 6 weeks after surgery. RESULTS: The SSN group reported significantly lower levels of postoperative pain, required significantly less analgesia, had better range of motion, and had higher levels of postoperative satisfaction in comparison to the subacromial infiltration group and placebo group. CONCLUSIONS: Patients treated with SSN blocks had less pain overall, which led to a decreased need for analgesics in comparison to the subacromial infiltration and placebo groups. Furthermore, patients in the SSN-blocked group achieved better postoperative ROM and were significantly more satisfied after surgery.

Synthesis and characterization of chiral benzylic ether-bridged periodic mesoporous organosilicas.

The first synthesis of a chiral periodic mesoporous organosilica (PMO) carrying benzylic ether bridging groups is reported. By hydrolysis and condensation of the new designed chiral organosilica precursor 1,4-bis(triethoxysilyl)-2-(1-methoxyethyl)benzene (BTEMEB) in the presence of the non-ionic oligomeric surfactant Brij 76 as supramolecular structure-directing agent under acidic conditions, an ordered mesoporous chiral benzylic ether-bridged hybrid material with a high specific surface area was obtained. The chiral PMO precursor was synthesized in a four-step reaction from 1,4-dibromobenzene as the starting compound. The evidence for the presence of the chiral units in the organosilica precursor as well as inside the PMO material is provided by optical activity measurements.

Carbonyl reductases and pluripotent hydroxysteroid dehydrogenases of the short-chain dehydrogenase/reductase superfamily.

Carbonyl reduction of aldehydes, ketones, and quinones to their corresponding hydroxy derivatives plays an important role in the phase I metabolism of many endogenous (biogenic aldehydes, steroids, prostaglandins, reactive lipid peroxidation products) and xenobiotic (pharmacologic drugs, carcinogens, toxicants) compounds. Carbonyl-reducing enzymes are grouped into two large protein superfamilies: the aldo-keto reductases (AKR) and the short-chain dehydrogenases/reductases (SDR). Whereas aldehyde reductase and aldose reductase are AKRs, several forms of carbonyl reductase belong to the SDRs. In addition, there exist a variety of pluripotent hydroxysteroid dehydrogenases (HSDs) of both superfamilies that specifically catalyze the oxidoreduction at different positions of the steroid nucleus and also catalyze, rather nonspecifically, the reductive metabolism of a great number of nonsteroidal carbonyl compounds. The present review summarizes recent findings on carbonyl reductases and pluripotent HSDs of the SDR protein superfamily.

[Arthroscopic Bankart operation using absorbable suture anchors].

OBJECTIVE: Arthroscopic refixation of the labrum-ligament complex at the glenoid. INDICATIONS: Posttraumatic anterior or anterior-inferior shoulder instability with Bankart or ALPSA lesion (anterior labral periosteal sleeve avulsion). CONTRAINDICATIONS: Atraumatic shoulder instability. Instabilities due to blunted or frayed degeneration of the labrum-ligament complex. HAGL lesion (humeral avulsion of the glenohumeral ligaments) with humeral detachment of the glenohumeral ligaments. Larger bony glenoid defects. SURGICAL TECHNIQUE: Mobilization of the labrum-ligament complex from the neck of the glenoid, superior tightening and refixation at the glenoid rim with the aid of absorbable suture anchors. POSTOPERATIVE MANAGEMENT: Immobilization of the affected arm for 4 weeks in an immobilization bandage with abduction pillows. Daily pendulum exercises. Active flexion up to 70 degrees and abduction up to 40 degrees, all in neutral or internal rotation. Avoidance of external rotation for a total of 6 weeks. RESULTS: From January 1999 to December 2001, 58 patients with a Bankart or ALPSA lesion were treated with arthroscopic shoulder stabilization using absorbable suture anchors and slowly absorbable braided sutures. 56 patients underwent a follow-up clinical examination after, on average, 31 months (24-48 months). None of these patients had suffered more than five shoulder dislocations before the operation (average 2.8). Of the intraoperative lesions, a plain Bankart lesion was present in twelve patients (21.4%), 44 patients had an ALPSA lesion (78.6%), of which one in two were combined with an SLAP 2 or SLAP 3 lesion (superior labrum from anterior to posterior). In the evaluation using the Rowe Score, there was an excellent result for 40 patients (71.4%), and a good result for twelve (21.4%). Four patients suffered a repeat dislocation and were therefore classified as poor results (7.2%).

Change of extracellular cAMP concentration is a sensitive reporter for bacterial fitness in high-cell-density cultures of Escherichia coli.

Guanosine-3',5'-tetraphosphate (ppGpp) and sigmaS, two regulators of the starvation response of Escherichia coli, have received increasing attention for monitoring cell physiological changes in production processes, although both are difficult to quantify. The kinetics of cAMP formation and degradation were not yet investigated in such processes, although the complex regulation of cAMP by synthesis, release, and degradation in connection with straightforward methods for analysis renders it a highly informative target. Therefore, we followed the cAMP concentration in various nonrecombinant and in four different recombinant glucose-limited fed-batch processes in different production scales. The intracellular cAMP concentration increases strongly at the end of the batch phase. Most cAMP is released to the cultivation medium. The rates of accumulation and degradation of extracellular cAMP are growth-rate-dependent and show a distinct maximum at a growth rate of about 0.35 h(-1). At very low growth rates, below 0.05 h(-1), extracellular cAMP is not produced but rather degraded, independent of whether this low growth rate is caused by glucose limitation or by the high metabolic load of recombinant protein production. In contrast to intracellular cAMP, which is highly unstable, analysis of extracellular cAMP is simpler and the kinetics of accumulation and degradation reflect well the physiological situation, including unlimited growth, limitation, and severe starvation of a production host.

Roles of heat-shock chaperones in the production of recombinant proteins in Escherichia coli.

Escherichia coli is a versatile organism for the production of recombinant proteins. Often, however, the recombinant protein does not reach its native, biologically active conformation within the bacterial cell but deposits as inclusion bodies. The heat-shock chaperones, a group of polypeptides omnipresent in all kingdoms of life, form a network to assist proper folding of cellular proteins, prevent their deposition and can even dissolve deposits of misfolded proteins formed during environmental stress conditions such as excessive heat. Coproduction of individual chaperones with the target protein can also reduce deposition of the recombinant protein into inclusion bodies. The selection of the suitable chaperone(s), however, is still a trial-and-error process. The wrong chaperone(s) will not lead to success, or may even negatively effect product stability or host viability. Recent progress in understanding the mechanisms and substrate specificities of the major chaperones and their roles in the chaperone network now gives some hints for a more rational choice of chaperone(s) for coproduction. Also, more specialized chaperone systems may become an alternative for application in the production of recombinant proteins.