Enrichment of cell cycle pathways in progesterone-treated endometrial organoids of infertile women compared to fertile women.

PURPOSE: To investigate whether the transcriptome profile differs between progesterone-treated infertile and fertile endometrial organoids. METHODS: Endometrial biopsies were obtained from 14 infertile and seven fertile women, after which organoids were generated from isolated epithelial cells. To mimic the secretory phase, organoids were sequentially treated with 17ß-estradiol (E2) and progesterone (P4) and subjected to RNA sequencing. Differentially expressed genes (DEGs) were identified using DESeq2 (lfcThreshold = 0, log2 Fold Change ≥ 1.0 or ≤ -1.0), and a principal component analysis (PCA) plot was generated. Functional enrichment analysis was performed by overrepresentation analysis and Gene Set Enrichment Analysis (GSEA). To functionally assess proliferation, OrganoSeg surface measurements were performed before (T0) and after (T1) differentiation of organoids, and T1/T0 ratios were calculated to determine the proliferation rate. RESULTS: Although the PCA plot did not show clear clustering of the fertile and infertile samples, 363 significant DEGs (129 upregulated and 234 downregulated) were detected in infertile compared to fertile organoids. Mainly cell cycle processes were highly enriched in infertile organoids. Thus, we hypothesised that proliferative activity during differentiation may be higher in infertile organoids compared to fertile organoids. However, this could not be validated by cell surface measurements. CONCLUSIONS: This study revealed that cell cycle processes were enriched in E2/P4-treated infertile endometrial organoids as compared to fertile organoids. This could reflect persistently higher proliferative activity of the endometrial epithelial cells in differentiated infertile organoids compared to fertile organoids. To confirm this hypothesis, further studies are warranted.

Who Benefited the Most? Effectiveness of a Lifestyle Intervention Against Cognitive Decline in Older Women and Men - Secondary Analysis of the AgeWell.de-trial.

INTRODUCTION: Differences between women and men matter in the prevalence and risk factors of dementia. We aimed to examine potential sex differences regarding the effectiveness by running a secondary analysis of the AgeWell.de trial, a cluster-randomized multicenter multi-domain lifestyle intervention to reduce cognitive decline. METHODS: Intention-to-treat analyses of women (n=433) and men (n=386) aged 60 to 77 years were used for models including interactions between intervention group allocation and sex followed by subgroup analysis stratified by sex on primary and secondary outcomes. Further, the same procedure was repeated for age groups (60-69 vs. 70-77) within sex-specific subgroups to assess the effectiveness in different age groups. TRIAL REGISTRATION: German Clinical Trials Register (ref. number: DRKS00013555). RESULTS: No differences were found between women and men in the effectiveness of the intervention on cognitive performance. However, women benefitted from the intervention regarding depressive symptoms while men did not. Health-related quality of life was enhanced for younger intervention participants (60-69 years) in both women and men. CONCLUSION: The AgeWell.de intervention was able to improve depressive symptoms in women and health-related quality of life in younger participants. Female participants between 60 and 69 years benefited the most. Results support the need of better individually targeted lifestyle interventions for older adults.

Prevention of legionella infections from toilet flushing cisterns.

INTRODUCTION: Immunocompromised patients are at an increased risk of severe legionella infections. We present the results of an outbreak investigation initiated following a fatal case of hospital-acquired legionellosis linked to contaminated water from a toilet-flushing cistern. Additionally, we provide experimental data on the growth of Legionella spp. in flushing cisterns and propose a straightforward protocol for prevention. METHODS: We monitored the growth of Legionella spp. in the building's hot- and cold-water systems using quantitative bacterial culture on selective agar. Molecular typing of Legionella pneumophila isolates from the infected patient and the water system was conducted through core-genome multi-locus sequence typing (cgMLST). RESULTS: Legionella contamination in the hospital building's cold-water system was significantly higher than in the hot-water system and significantly higher in toilet flushing cistern's water compared with cold water from bathroom sinks and showers. Isolates from the patient and from the flushing cistern of the patient's bathroom were identical by cgMLST. In an experimental setting, daily toilet flushing for a period of 21 days resulted in a 67% reduction in the growth of Legionella spp. in the water of toilet flushing cisterns. Moreover, a one-time disinfection of cisterns with peracetic acid, followed by daily flushing, decreased legionella growth to less than 1% over a period of at least seven weeks in these setting. CONCLUSIONS: One-time disinfection of highly contaminated cisterns with peracetic acid and daily toilet flushing as short-term measure can significantly reduce legionella contamination in flushing cisterns. These measures may aid in preventing legionella infection among immunocompromised patients.

Vibrationally Resolved Inner-Shell Photoexcitation of the Molecular Anion C2.

Carbon 1s core-hole excitation of the molecular anion C2 - has been experimentally studied at high resolution by employing the photon-ion merged-beams technique at a synchrotron light source. The experimental cross section for photo-double-detachment shows a pronounced vibrational structure associated with 1 σ u → 3 σ g ${1\sigma _u \to 3\sigma _g }$ and 1 σ g → 1 π u ${1\sigma _g \to 1\pi _u }$ core excitations of the C2 - ground level and first excited level, respectively. A detailed Franck-Condon analysis reveals a strong contraction of the C2 - molecular anion by 0.2 Šupon this core photoexcitation. The associated change of the molecule's moment of inertia leads to a noticeable rotational broadening of the observed vibrational spectral features. This broadening is accounted for in the present analysis which provides the spectroscopic parameters of the C2 - 1 σ u - 1 3 σ g 2 2 Σ u + ${1\sigma _u^{ - 1} \,3\sigma _g^2 \;^2 \Sigma _u^ + }$ and 1 σ g - 1 3 σ g 2 2 Σ g + ${1\sigma _g^{ - 1} \,3\sigma _g^2 \;^2 \Sigma _g^ + }$ core-excited levels.

[Removal of a bent femoral nail : Case report: treatment strategy of a complex femoral deformity with a bent femoral nail in situ since 36 years]. / Entfernung eines verbogenen Femurnagels : Kasuistik: Behandlungsstrategie einer komplexen Femurdeformität mit einem seit 36 Jahren einliegenden, verbogenen Femurnagel.

To remove a bent femoral nail is always a challenging task for a surgeon. In the last 20 years several case reports and descriptions of techniques are to be found in literature, the goal being to weaken the nail at the apex and then straightening it in situ in most cases. Those reports all have one parameter in common: the necessity to remove the nail relates to a refracture of the femur.We report the case of a patient who presented with the explicit wish to have a bent femoral nail removed and a femoral angulation corrected because of pain in the ipsilateral hip. A femoral fracture had been treated by a Küntscher nail in 1982. A refracture with severe bending of the nail had occurred a short time later, which was treated conservatively without a second surgical procedure.We can show that a planned removal of a bent and well-osseointegrated Küntscher nail can be achieved without complications and that thereby a multiplane angulation in the femur can be corrected, paving the way for a future arthroplasty in this specific case.

A wildfire vulnerability index for buildings.

Recent wildfire events (e.g. Mediterranean region, USA, and Australia) showed that this hazard poses a serious threat for wildland-urban interface (WUI) areas around the globe. Furthermore, recent events in regions where wildfire does not constitute a frequent hazard (e.g. Siberia, Scandinavia) indicated that the spatial pattern of wildfire risk might have significantly changed. To prepare for upcoming extreme events, it is critical for decision-makers to have a thorough understanding of the vulnerability of the built environment to wildfire. Building quality and design standards are important not only because building loss is costly but also because robust buildings may offer shelter when evacuation is not possible. However, studies aiming at the analysis of wildfire vulnerability for the built environment are limited. This paper presents an innovative solution for the vulnerability assessment to wildfires, making use of an all-relevant feature selection algorithm established on statistical relationships to develop a physical vulnerability index for buildings subject to wildfire. Data from a recent and systematically documented wildfire event in Greece (Mati, 2018) are used to select and weight the relevant indicators using a permutation-based automated feature selection based on random forests. Building characteristics including the structural type, the roof type, material and shape, the inclination of the ground, the surrounding vegetation, the material of the shutters and the ground covering were selected and formed into the index. The index may be used in other places in Europe and beyond, especially where no empirical data are available supporting decision-making and risk reduction of an emerging hazard amplified by climate change.

Mutation-specific reporter for optimization and enrichment of prime editing.

Prime editing is a versatile genome-editing technique that shows great promise for the generation and repair of patient mutations. However, some genomic sites are difficult to edit and optimal design of prime-editing tools remains elusive. Here we present a fluorescent prime editing and enrichment reporter (fluoPEER), which can be tailored to any genomic target site. This system rapidly and faithfully ranks the efficiency of prime edit guide RNAs (pegRNAs) combined with any prime editor variant. We apply fluoPEER to instruct correction of pathogenic variants in patient cells and find that plasmid editing enriches for genomic editing up to 3-fold compared to conventional enrichment strategies. DNA repair and cell cycle-related genes are enriched in the transcriptome of edited cells. Stalling cells in the G1/S boundary increases prime editing efficiency up to 30%. Together, our results show that fluoPEER can be employed for rapid and efficient correction of patient cells, selection of gene-edited cells, and elucidation of cellular mechanisms needed for successful prime editing.

Comparison of novel approaches for expedited pathogen identification and antimicrobial susceptibility testing against routine blood culture diagnostics.

Blood stream infections pose a major challenge for clinicians as the immediate application of an appropriate antibiotic treatment is the vital factor to safe the patients' lives. This preliminary study compares three different systems promising fast pathogen identification and susceptibility testing in comparison to conventional blood culture (BC): (i) the rapid antimicrobial susceptibility testing protocol according to EUCAST in combination with the Sepsityper® kit (sRAST), (ii) the direct inoculation method on the VITEK® 2 system (dVIT) and (iii) testing with the Accelerate Pheno® system (AccPh). All methods were assessed in terms of accuracy, time to result and usability. Twenty-three BC samples obtained from patients suffering from proven sepsis were analysed in detail. Pathogen identification was successful in 95·6, 91·3 and 91·3% in sRAST, dVIT and AccPh, respectively. Categorical agreement in antimicrobial susceptibility testing was 89·5, 96 and 96·6%, respectively. Time to result from sample entry to reporting ranged from an average of 4·6 h for sRAST and 6·9 h for AccPh to 10·6 h for dVIT. These results imply a significant shortening of reporting times at considerably high agreement rates for these new diagnostic approaches.

[Obesity and eating disorders]. / Vignette thérapeutique de l'étudiant. Obésité et troubles du comportement alimentaire.

Obesity is a chronic disease that is difficult to manage and is often punctuated by treatment failure. Screening for eating disorders, the prevalence of which is high in overweight patients, is essential in order to minimize the risk of relapse and the «yoyo¼ phenomenon. As the reason for consulting obese subjects is rare to be eating disorders from the outset, it is necessary to assess this behaviour from the first consultations, in order to propose a specific therapeutic approach aimed at stable weight loss in the long term.

L'obésité est une maladie chronique dont la prise en charge est ardue et souvent ponctuée d'échecs. Le dépistage des troubles du comportement alimentaire, dont la prévalence est élevée chez les patients en surpoids, est indispensable afin de minimiser le risque de rechute et le phénomène «yoyo¼. Comme il est rare que le motif de consultation des sujets souffrant d'obésité soit d'emblée le désordre alimentaire, il est nécessaire d'évaluer ce comportement, dès les premières consultations, afin de proposer une approche thérapeutique spécifique, visant une perte de poids stable au long cours.

High protein prescription in methylmalonic and propionic acidemia patients and its negative association with long-term outcome.

BACKGROUND AND OBJECTIVE: Methylmalonic acidemia (MMA) and propionic acidemia (PA) are inborn errors of metabolism. While survival of MMA and PA patients has improved in recent decades, long-term outcome is still unsatisfactory. A protein restricted diet is the mainstay for treatment. Additional amino acid mixtures (AAM) can be prescribed if natural protein is insufficient. It is unknown if dietary treatment can have an impact on outcome. DESIGN: We performed a nationwide retrospective cohort study and evaluated both longitudinal dietary treatment and clinical course of Dutch MMA and PA patients. Protein prescription was compared to the recommended daily allowances (RDA); the safe level of protein intake as provided by the World Health Organization. The association of longitudinal dietary treatment with long-term outcome was evaluated. RESULTS: The cohort included 76 patients with a median retrospective follow-up period of 15 years (min-max: 0-48 years) and a total of 1063 patient years on a protein restricted diet. Natural protein prescription exceeded the RDA in 37% (470/1287) of all prescriptions and due to AAM prescription, the total protein prescription exceeded RDA in 84% (1070/1277). Higher protein prescriptions were associated with adverse outcomes in severely affected patients. In PA early onset patients a higher natural protein prescription was associated with more frequent AMD. In MMA vitamin B12 unresponsive patients, both a higher total protein prescription and AAM protein prescription were associated with more mitochondrial complications. A higher AAM protein prescription was associated with an increased frequency of cognitive impairment in the entire. CONCLUSION: Protein intake in excess of recommendations is frequent and is associated with poor outcome.

Interlaboratory evaluation of Mucorales PCR assays for testing serum specimens: A study by the fungal PCR Initiative and the Modimucor study group.

Interlaboratory evaluations of Mucorales qPCR assays were developed to assess the reproducibility and performance of methods currently used. The participants comprised 12 laboratories from French university hospitals (nine of them participating in the Modimucor study) and 11 laboratories participating in the Fungal PCR Initiative. For panel 1, three sera were each spiked with DNA from three different species (Rhizomucor pusillus, Lichtheimia corymbifera, Rhizopus oryzae). For panel 2, six sera with three concentrations of R. pusillus and L. corymbifera (1, 10, and 100 genomes/ml) were prepared. Each panel included a blind negative-control serum. A form was distributed with each panel to collect results and required technical information, including DNA extraction method, sample volume used, DNA elution volume, qPCR method, qPCR template input volume, qPCR total reaction volume, qPCR platform, and qPCR reagents used. For panel 1, assessing 18 different protocols, qualitative results (positive or negative) were correct in 97% of cases (70/72). A very low interlaboratory variability in Cq values (SD = 1.89 cycles) were observed. For panel 2 assessing 26 different protocols, the detection rates were high (77-100%) for 5/6 of spiked serum. There was a significant association between the qPCR platform and performance. However, certain technical steps and optimal combinations of factors may also impact performance. The good reproducibility and performance demonstrated in this study support the use of Mucorales qPCR as part of the diagnostic strategy for mucormycosis.

Cluedo - Source identification in a case of septicemia fatality caused by Capnocytophaga canimorsus.

The Gram negative pathogen Capnocytophaga canimorsus is a frequent commensal in the oral cavity of cats and dogs. Although the bacterium is generally considered harmless, infections in humans can occur displaying a broad spectrum of clinical syndromes. This makes a clinical diagnosis difficult. The patient in the present case was 67 years old and presented to the emergency room (ER) with pain in the upper right abdomen and clinical signs of a feverish infection. The only noticeable record in the patient´s medical history was a splenectomy in childhood. The anamnesis revealed that the patient was the owner of two dogs. After a suspected diagnosis of sepsis the patient was transferred to the intensive care unit (ICU), where his medical condition deteriorated rapidly. Despite intensive care measures as well as the fast initialization of a broad-spectrum antibiotic therapy, the patient died 37 h after his presentation in the ER. The search for the causative pathogen turned out to be challenging. Eventually, molecular biological methods assisted in solving the puzzle. It could be demonstrated that the pathogen, found in the patient´s blood, was also present in one of his dogs' oral cavity.

Incidence, predictors and prognostic implications of dyspnea at admission among acute coronary syndrome patients without heart failure.

BACKGROUND: Among patients with stable coronary artery disease, effort-related dyspnea is associated with a larger ischemic territory and worse outcome. Whether dyspnea, not related to heart failure, is also associated with adverse outcome among patients with acute coronary syndromes (ACS) has not been fully elucidated. METHODS: We studied ACS patients enrolled in the biennial Acute Coronary Syndrome Israeli Surveys (ACSIS) during 2010-2013 who were classified as Killip 1. A retrospective comparative analysis was performed between patients with chest pain alone (n = 2017) and those with chest pain with dyspnea (n = 417). RESULTS: Patients with dyspnea were older (64.4 ±â€¯13 vs.61.8 ±â€¯12, p < 0.001), more frequently women (81% vs. 75% p < 0.001) and had higher rates of multiple comorbidities. Statistically significant predictors for dyspnea as a presenting symptom were female sex [HR 1.47 (1.11, 1.89)], chronic kidney disease [HR 1.81 (1.30, 2.52)], chronic obstructive pulmonary disease [HR 1.59 (1.045, 2.429)] and angina ≥24 h [HR 1.46 (1.147, 1.86)]. Patients presenting with dyspnea were less likely to undergo primary reperfusion (31% vs. 42%, p < 0.001) and overall coronary intervention (71% vs. 78%, p < 0.001) during their hospitalization. Mortality rates were significantly higher among patients presenting with dyspnea both at 30-day (3% vs. 2%, p = 0.017) and at 1-year follow-up (9% vs. 4%, p < 0.001). Dyspnea was as an independent predictor of 1-year mortality. CONCLUSION: The presence of dyspnea is frequent and associated with adverse outcome among patients with ACS without signs of heart failure. Early identification of this higher-risk cohort of patients may allow intensifying treatment and careful follow-up may be warranted.

[Uveal Melanoma Cell Under Oxidative Stress - Influence of VEGF and VEGF-Inhibitors]. / Uveale Melanomzellen unter oxidativem Stress ­ Einfluss von VEGF und VEGF-Inhibitoren.

BACKGROUND: The role of oxidative stress in cancer is complex. While the pathological alterations induced by oxidative stress may be involved in the induction of tumours, in the late stages of tumour development, it can facilitate the loss of tumour cells and might even prevent metastasis. Tumour cells show metabolic alterations, often inducing an increased production of reactive oxygen species, which makes these cells particularly vulnerable to additional oxidative stress. This is an important mode of action in the use of many chemotherapeutics and in the application of ionizing radiation. Uveal melanoma is the most frequent primary tumour in the adult eye. For metastasis of this tumour, which affects about 50 % of the patients, no appropriate treatment is currently available. However, the primary tumour can efficiently be treated with ionizing radiation. A frequent side effect of this treatment is radiation retinopathy, which is treated with vascular endothelial growth factor (VEGF) antagonists. A therapy of the primary tumour with VEGF antagonists is under discussion. So far, little data is available on this subject, however, a paradoxical worsening of the situation has been found in a mouse model of uveal melanoma treated with bevacizumab. METHODS: We have investigated the effect of VEGF and of the VEGF-antagonist bevacizumab on the survival of five different melanoma cell lines under oxidative stress treatment with hydrogen peroxide. In addition, we investigated the expression of relevant proteins and the effect of bevacizumab on the proliferation of the cells as well as its effect on the angiogenic behaviour of endothelial cells, co-cultured with uveal melanoma cells. RESULTS: Our study showed that not only VEGF but also, paradoxically, the VEGF-antagonist bevacizumab is able to protect uveal melanoma cells from oxidative stress-induced cell death. Bevacizumab did not influence the proliferation of the cells and showed only limited effectiveness to reduce angiogenic structures. CONCLUSION: Considering that oxidative stress is the mode of action for ionizing radiation to induce cell death, a protective effect of bevacizumab on uveal melanoma cells against oxidative stress is worrisome and argues against the use of VEGF in uveal melanoma.

Unraveling the Nature of Magnetism of the 5d

We report electron spin resonance (ESR) spectroscopy results on the double perovskite Ba_{2}YIrO_{6}. On general grounds, this material is expected to be nonmagnetic due to the strong coupling of the spin and orbital momenta of Ir^{5+} (5d^{4}) ions. However, controversial experimental reports on either strong antiferromagnetism with static order at low temperatures or just a weakly paramagnetic behavior have triggered a discussion on the breakdown of the generally accepted scenario of the strongly spin-orbit coupled ground states in the 5d^{4} iridates and the emergence of a novel exotic magnetic state. Our data evidence that the magnetism of the studied material is solely due to a few percent of Ir^{4+} and Ir^{6+} magnetic defects while the regular Ir^{5+} sites remain nonmagnetic. Remarkably, the defect Ir^{6+} species manifest magnetic correlations in the ESR spectra at T≲20 K, suggesting a long-range character of superexchange in the double perovskites as proposed by recent theories.

Microsolvation of phthalocyanine molecules in superfluid helium nanodroplets as revealed by the optical line shape at electronic origin.

We investigate the solvent shift of phthalocyanine (Pc) doped into superfluid helium droplets and probed by optical spectroscopy at the electronic origin. Our present work complements extant studies and provides results that in part contradict previous conclusions. In particular, the solvent shift does not increase monotonously with droplet radius all the way up to the bulk limit, but exhibits just the reverse dependence instead. Moreover, a substructure is resolved, whose characteristics depend on the droplet size. This behavior can hardly be reconciled with that of a freely rotating Pc-helium complex.

Microsolvation of porphine molecules in superfluid helium nanodroplets as revealed by optical line shape at the electronic origin.

We investigate the line shape at the electronic origin of single porphine molecules doped into superfluid helium droplets as a function of the droplet size. Helium droplets comprised of less than 105 atoms are generated from an expansion of gaseous helium, while droplets with more than 105 atoms originate from liquid helium. In contrast to our recent study on phthalocyanine, porphine is found to exhibit a solvent shift to the blue with respect to the gas-phase resonance frequency as well as a multiplet splitting. A comparison of the helium-induced features of phthalocyanine and porphine with those obtained in similar studies on tetracene and pentacene reveals that these occur chiefly as two kinds of excitations distinguished by their linewidths and their dependence on the droplet size. Moreover, at quasi-bulk conditions achieved with droplets in excess of 106 helium atoms, none of these four dopant species yields an optical spectrum that can be assigned to a plausible rotational band structure.

Cross-sectional point prevalence survey to study the environmental contamination of nosocomial pathogens in intensive care units under real-life conditions.

BACKGROUND: In intensive care units (ICUs), inanimate surfaces and equipment may be contaminated by nosocomial pathogens, including multi-drug-resistant micro-organisms. AIMS: To assess the degree of environmental contamination close to and distant from patients, and contamination of healthcare workers' (HCWs) hands with nosocomial pathogens under real-life conditions and to investigate potential transmission events. METHODS: Over the course of three weeks, agar contact samples were taken close to and distant from patient areas and from HCWs' hands in eight ICUs of a tertiary care hospital in Innsbruck, Austria. Each ICU was visited once without announcement. Species identification and antimicrobial susceptibility testing were performed according to standard methods, and corresponding strains from patient, environment and hand samples were genotyped using pulsed-field gel electrophoresis. FINDINGS: Among 523 samples, HCWs' hands were most frequently contaminated with potentially pathogenic bacteria (15.2%), followed by areas close to patients (10.9%) and areas distant from patients (9.1%). Gram-positive bacteria were identified most often (67.8%), with Enterococcus spp. being the most prevalent species (70% vancomycin sensitive and 30% vancomycin resistant) followed by Staphylococcus aureus, of which 64% were classified as meticillin-resistant Staphylococcus aureus. Molecular typing documented identical strains among patient, environment and hand isolates. CONCLUSION: This study found widespread contamination of the ICU environment with clinically relevant pathogens, including multi-drug-resistant micro-organisms, despite cleaning and disinfection. The bioburden might not be restricted to areas close to patients. The role of extended environmental disinfection of areas distant from patients in order to improve infection prevention needs further discussion.

Serum netrin-1 in relation to gadolinium-enhanced magnetic resonance imaging in early multiple sclerosis.

BACKGROUND: Netrin-1, a secreted laminin-related protein, is known to regulate not only axonal guidance and neuronal cell migration, but also blood-brain barrier integrity and inflammation. Two preliminary studies reported altered serum netrin-1 levels in multiple sclerosis; however, associations with longitudinal clinical and magnetic resonance imaging activity have not been investigated. OBJECTIVES: We aimed to assess serum netrin-1 in multiple sclerosis and controls with respect to disease activity and its temporal dynamics. METHODS: Serum netrin-1 was assessed by enzyme-linked immunosorbent assay in 79 patients with clinically isolated syndrome or multiple sclerosis, and 30 non-inflammatory neurological disease controls. In patients, serum samples were collected immediately prior to gadolinium-enhanced 3 T magnetic resonance imaging at two time points (initial contrast-enhancing gadolinium+ n = 47, non-enhancing gadolinium- n = 32; reference gadolinium- n = 70; median time-lag 1.4, interquartile range 1.0-2.3 years). RESULTS: Serum netrin-1 levels were similar in clinically isolated syndrome, multiple sclerosis and controls, and gadolinium+ and gadolinium- patients. Among gadolinium+ patients, serum netrin-1 was decreased in clinically active (n = 8) vs non-active patients (n = 39; p = 0.041). Serum netrin-1 showed no temporal dynamics in multiple sclerosis and was unrelated to clinical data. CONCLUSIONS: Serum netrin-1 levels show no multiple sclerosis specific changes and are not sensitive for detection of subclinical disease activity. Netrin-1 changes during relapses may deserve further examination.

Use of a gamma-2 herpesvirus as a vector to deliver antibodies to rhesus monkeys.

The gamma-2 herpesvirus of rhesus monkeys, rhesus monkey rhadinovirus (RRV), persists principally in B cells of its host. We constructed recombinant strains of RRV expressing the rhesus monkey-derived anti-SIV monoclonal antibodies 4L6 and 5L7 and compared the RRV-mediated in vivo delivery of these antibodies in rhesus monkeys with previous studies that utilized intramuscular delivery with an adeno-associated virus (AAV) vector. Recombinant RRV-4L6 and RRV-5L7 were both shown to stably produce the antibodies in persistently infected B-cell lines in culture. Two RRV-negative rhesus monkeys were experimentally infected with recombinant RRV-4L6 and two with recombinant RRV-5L7. Following infection, the appearance of the delivered antibody was readily detected in all four animals. However, the levels of the delivered antibody were considerably lower than what has been typically observed following intramuscular AAV delivery. Furthermore, three of the four monkeys had an antibody response to the delivered antibody as had been observed previously with intramuscular AAV delivery of these same antibodies. We conclude that this recombinant herpesvirus has no inherent advantage over AAV for delivery of potentially therapeutic monoclonal antibodies in a rhesus monkey model.