Advanced optoacoustic methods for multiscale imaging of in vivo dynamics.

Visualization of dynamic functional and molecular events in an unperturbed in vivo environment is essential for understanding the complex biology of living organisms and of disease state and progression. To this end, optoacoustic (photoacoustic) sensing and imaging have demonstrated the exclusive capacity to maintain excellent optical contrast and high resolution in deep-tissue observations, far beyond the penetration limits of modern microscopy. Yet, the time domain is paramount for the observation and study of complex biological interactions that may be invisible in single snapshots of living systems. This review focuses on the recent advances in optoacoustic imaging assisted by smart molecular labeling and dynamic contrast enhancement approaches that enable new types of multiscale dynamic observations not attainable with other bio-imaging modalities. A wealth of investigated new research topics and clinical applications is further discussed, including imaging of large-scale brain activity patterns, volumetric visualization of moving organs and contrast agent kinetics, molecular imaging using targeted and genetically expressed labels, as well as three-dimensional handheld diagnostics of human subjects.

T-cell-biased immune responses generated by a mucosally targeted adenovirus-σ1 vaccine.

As most pathogens enter through the mucosa, it is important to develop vaccines that induce mucosal immunity. To this end, we generated a novel adenovirus (Ad) vaccine that displays the σ1 protein from reovirus to target junctional adhesion molecule 1 and sialic acid. Replication-defective Ad5 vectors were modified by replacement of the Ad fiber protein with σ1 (T3Dσ1) protein of reovirus T3D in previous work. Ad5 and Ad5-σ1 were compared in mouse models for gene delivery and vaccination to monitor cytokine, antibody, and T-cell responses. The viruses were also tested for the ability to transduce and mature dendritic cells. Ad5-σ1 was 40-fold less efficient at gene delivery in vivo, yet it was capable of inducing equal or greater cellular immune responses and systemic interferon-γ levels than Ad5 after intranasal administration. Despite weaker gross transduction, intranasal administration of Ad5-σ1 produced more green fluorescent protein-positive (GFP+) major histocompatibility complex class II (MHC II) cells in the draining lymph nodes, less GFP+/MHC II+ cells in the lungs, and mediated modestly better maturation of dendritic cells in vitro. These data suggest that targeting gene-based vaccination via the σ1 protein may enhance the T-cell immune response, perhaps by skewing immune responses to encoded antigens.

A phase II study evaluating the safety and efficacy of an adenovirus-ΔLMP1-LMP2 transduced dendritic cell vaccine in patients with advanced metastatic nasopharyngeal carcinoma.

BACKGROUND: Individuals with metastatic Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) continue to have poor outcomes. To evaluate the ability of a dendritic cell (DC) vaccine to target subdominant EBV antigens LMP1 and LMP2 expressed by NPC cells, we vaccinated patients using autologous DCs transduced with an adenovirus encoding a truncated LMP1 (ΔLMP1) and full-length LMP2 (Ad-ΔLMP1-LMP2). MATERIALS AND METHODS: Sixteen subjects with metastatic NPC received Ad-ΔLMP1-LMP2 DC vaccines i.d. biweekly for up to five doses. Toxicity, immune responses and clinical responses were determined. RESULTS: Most patients had extensive disease, with a median of three visceral sites of involvement (range 1-7). No significant toxicity was observed. Ad-ΔLMP1-LMP2 DCs induced delayed type hypersensitivity responses in 9 out of 12 patients, but although these DCs activated LMP1/2-specific T cells in vitro, no such increase in the frequency of peripheral LMP1/2-specific T cells was detected. Three patients had clinical responses including one with partial response (for 7½ months) and two with stable disease (for 6½ and 7½ months). CONCLUSIONS: Ad-ΔLMP1-LMP2 transduced DCs can be successfully generated and safely administered to patients with advanced NPC. Since efficacy was limited, future studies should focus on DC vaccines with greater potency administered to subjects with less tumor burden.

Immunotherapy targeting HER2 with genetically modified T cells eliminates tumor-initiating cells in osteosarcoma.

Despite radical surgery and multi-agent chemotherapy, less than one third of patients with recurrent or metastatic osteosarcoma (OS) survive. The limited efficacy of current therapeutic approaches to target tumor-initiating cells (TICs) may explain this dismal outcome. The purpose of this study was to assess the impact of modified T cells expressing a human epidermal growth factor receptor (HER2)-specific chimeric antigen receptor in the OS TIC compartment of human established cell lines. Using the sarcosphere formation assay, we found that OS TICs were resistant to increasing methotrexate concentrations. In contrast, HER2-specific T cells decreased markedly sarcosphere formation capacity and the ability to generate bone tumors in immunodeficient mice after orthotopic transplantation. In vivo, administration of HER2-specific T cells significantly reduced TICs in bulky tumors as judged by decreased sarcosphere forming efficiency in OS cells isolated from explanted tumors. We demonstrate that HER2-specific T cells target drug resistant TICs in established OS cell lines, suggesting that incorporating immunotherapy into current treatment strategies for OS has the potential to improve outcomes.

Generating CTLs against the subdominant EBV LMP antigens by transient expression of an A20 inhibitor with EBV LMP proteins in human DCs.

Epstein-Barr virus (EBV) infection leads to Hodgkin's disease (HD) in some immunocompetent hosts. The malignant Reed-Sternberg cells of HD only express a limited array of subdominant EBV antigens to evade pre-existing immune responses to EBV. The EBV-encoded latent membrane proteins (LMP1 and LMP2), which are expressed by HD and various EBV-associated malignancies, have been proposed as a potential target for cytotoxic T lymphocyte (CTL)-based therapy. However, the precursor frequency for LMP-specific CTL is generally low in healthy EBV-infected hosts, and immunotherapy based on these antigens is often compromised by the poor immunogenicity and the oncogenic potential. In the present study, we report that transiently expressing an inhibitor of A20, a key negative regulator of inflammatory signaling pathways, together with the LMP antigens (truncated LMP1 and full-length LMP2) greatly enhances maturation and cytokine production of human (h) monocyte-derived dendritic cells (DCs). As a consequence, LMP1/2-expressed, A20-silenced hDCs have an enhanced potency to prime LMP-specific T-cell response. When the in vitro primed T cells are adoptively transferred into tumor-xenografted, severe-combined immunodeficient mice, some of the xenografted tumors approach complete regression. Thus, the study may provide an available resource of LMP-specific T cells for T-cell immunotherapy.

Isolated lympho-histiocytic inflammation of the Gasserian ganglion mimicking trigeminal schwannoma.

UNLABELLED: We present an unusual case of a patient with trigeminal pain and hypesthesia suggesting trigeminal schwannoma, where histology demonstrated an inflammatory lesion of the Gasserian ganglion. CLINICAL PRESENTATION: Patient history and imaging were typical for a trigeminal schwannoma with a dumbbell growth in the middle and posterior fossa. Surgery was performed and resection of a firm mass intermingled with fascicles originating from the ganglion and the mandibular branch was carried out via a subtemporal approach. Histological diagnosis revealed a non-caseating, vaguely granulomatous lymphocyte- and histiocyte-rich inflammation. CONCLUSION: Inflammatory lesions of the Gasserian ganglion are rare but may mimic tumors. Intraoperative biopsies should be taken prior to complete resection to limit neurological deterioration.

Outcome of hematopoietic stem cell transplant as salvage therapy for Hodgkin's lymphoma in adolescents and young adults at a single institution.

For patients with relapsed Hodgkin's lymphoma (HL), high dose chemotherapy with stem cell rescue (HDCT-SCT) may improve survival over chemotherapy alone. We assessed the outcomes of HDCT-SCT in 37 consecutive adolescent and young adult patients with relapsed HL whose malignancy was categorized based on sensitivity to chemotherapy. We determined whether current outcomes supported the use of HDCT-SCT in all of our patients or just those patients with lower-risk characteristics such as chemosensitivity. With a median follow-up of 6.5 years, the 2-year overall survival (OS) was 89% (95% CI: 62-97%) for the chemosensitive patients (n = 21), whereas for patients with resistant disease (n = 16), OS was 53% (95% CI: 25-74%). Both autologous and allogeneic transplants were well tolerated, with 100-day treatment-related mortality under 10%. Our data show encouraging outcomes for patients with chemosensitive relapsed HL who receive hematopoietic stem cell transplant (HSCT) and support the value of the procedure even when the disease is chemoresistant.

Vergence deficits in patients with cerebellar lesions.

The cerebellum is part of the cortico-ponto-cerebellar circuit for conjugate eye movements. Recent animal data suggest an additional role of the cerebellum for the control of binocular alignment and disconjugate, i.e. vergence eye movements. The latter is separated into two different components: fast vergence (to step targets) and slow vergence (to ramp and sinusoidal targets). The aim of this study was to investigate whether circumscribed cerebellar lesions affect these dynamic vergence eye movements. Disconjugate fast and slow vergence, conjugate smooth pursuit and saccades were binocularly recorded by a scleral search coil system in 20 patients with acute cerebellar lesions (all ischemic strokes except for one) and 20 age-matched healthy controls. Patients showed impairment of slow vergence while fast vergence was unaffected. Slow vergence gain to sinusoidal targets was significantly reduced, both in convergence and divergence direction. Divergence but not convergence velocity to ramp targets was reduced. Conjugate smooth pursuit eye movements to sinusoidal and to step-ramp targets were impaired. Patients had saccadic hypometria. All defects were particularly expressed in patients with vermis lesions. In contrast to recent animal data fast vergence was not impaired in any of our patient subgroups. We conclude that (i) the human cerebellum, in particular the vermis, is involved in the processing of dynamic vergence eye movements and (ii) cerebellar lesions elicit dissociable effects on fast and slow vergence.

[Posttraumatic gusher phenomenon following fracture of the petrous bone]. / Posttraumatisches Gusher-Phänomen nach Felsenbeinquerfraktur.

The gusher phenomenon is a very rare complication that may occur during stapedectomy or cochleostomy. A sudden perilymphatic flow of cerebrospinal fluid can be seen following platinotomy. The cause is an abnormal connection between subarachnoid and perilymphatic spaces due to congenital malformation, leading to an abnormally wide cochlear aqueduct or due to an internal auditory canal fistula. We describe a case of posttraumatic gusher phenomenon after a fracture of the petrous bone.

A nanoparticulate indium tin oxide field-effect transistor with solid electrolyte gating.

Reversible tuning of the transport properties of metallic conducting systems is not reported widely in the literature. Here, we report a junction field-effect transistor (FET) based on a transparent conducting oxide (TCO) nanoparticle channel and a solid polymer electrolyte as a gate. The device principle is based on the variation of the drain current induced by the capacitive double layer charging at the electrolyte/nanoparticle interfaces. A device with a metallic conducting channel made of indium tin oxide (ITO) nanoparticles exhibits an on/off ratio of 2 × 10(3) even when the gate potential is limited within the electrochemical capacitive region to avoid redox reactions at the interface. An FET device with metal-like conductance is always favored for the low dimensions of the device and a high on-state current. The field-effect mobility is calculated to be 24.3 cm(2) V(-1) s(-1). A subthreshold swing between 230 and 425 mV dec(-1) is observed.

[Simultaneous presentation of amyotrophic lateral sclerosis and multiple sclerosis]. / Kombiniertes Auftreten von amyotropher Lateralsklerose und Multipler Sklerose.

The concurrence of amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS) is extremely rare. We report on a 55-year-old female patient presenting with a progressive gait disorder for 6 months and a speech disorder for 3 months. Neurological examination revealed a spastic paraparesis and mild dysarthria and dysphagia. Technical and laboratory investigations met the diagnostic criteria for MS: magnetic resonance imaging showed multiple periventricular white matter and cervical lesions; cerebrospinal fluid showed a typical autoimmune response. Within the following 3 months generalized fasciculations, atrophy of the small hand muscles and bulbar signs were noticed. Nerve conduction studies revealed acute and chronic signs of denervation in all limbs without nerve conduction block. Hence clinical and paraclinical examination met the El Escorial criteria for ALS. Although myelitic lesions in the anterior horn cells may lead to peripheral segmental denervation, the generalized denervation suggested the unusual coincident combination of ALS and MS in this patient. In clinical praxis motoneuron diseases should also be considered in patients with pronounced peripheral denervations once "definite" MS has been diagnosed.

[Squamous cell carcinoma of the conjunctiva with orbital invasion: orbital exenteration or minimally invasive procedure?]. / Plattenepithelkarzinom der Bindehaut mit Orbitainvasion : Exenteratio orbitae oder minimal-invasives Vorgehen?

BACKGROUND: Here we present a case of a female patient with squamous cell carcinoma with orbital invasion who insisted on a globe-sparing excisional biopsy rather than a more invasive procedure. CASE: A 48-year-old patient presented with a conjunctival tumor at the medial canthus of the right eye. The visual acuity in this eye was 1.0. MRI showed a 25 mm x 12 mm tumor, with orbital invasion, no metastasis, but on CT scan damage of the lamina papyracea was suspected. Histology of a punch biopsy confirmed a moderately differentiated, squamous cell carcinoma. As requested by the patient, simple tumor excision with a 5-mm safety margin, reconstruction of the lower fornix with amniotic membrane, and adjuvant mitomycin C therapy were performed. Intraoperatively the medial orbital wall was found to be intact. No local or systemic recurrence has occurred 3.5 years postoperatively. The patient's VA is still 1.0; no visual field defects are detectable, and there are only minor upper lid levator restriction and diplopia in maximum upgaze. CONCLUSION: In view of this case the choice of surgical treatment for malignant tumors of the conjunctiva with orbital invasion - between exenteration or a minimally invasive procedure - should be carefully discussed with the patient. A follow-up of 5 years is strongly recommended.

Disseminated mycobacterium bovis infection in an immunocompetent host.

We report about a rare case of disseminated Mycobacterium bovis infection in a 61 year old female immunocompetent patient with involvement of the lung, the brain, the spleen and spine. The patient had intracerebral tuberculomas with paradoxical enlargement during the first weeks of therapy. We reviewed the data of our microbiological department and found five other patients with Mycobacterium bovis infection diagnosed between 1999 and 2004, which are 5.8 % of all diagnoses of tuberculosis during this period.

Inverse free electron lasers and laser wakefield acceleration driven by CO2 lasers.

The staged electron laser acceleration (STELLA) experiment demonstrated staging between two laser-driven devices, high trapping efficiency of microbunches within the accelerating field and narrow energy spread during laser acceleration. These are important for practical laser-driven accelerators. STELLA used inverse free electron lasers, which were chosen primarily for convenience. Nevertheless, the STELLA approach can be applied to other laser acceleration methods, in particular, laser-driven plasma accelerators. STELLA is now conducting experiments on laser wakefield acceleration (LWFA). Two novel LWFA approaches are being investigated. In the first one, called pseudo-resonant LWFA, a laser pulse enters a low-density plasma where nonlinear laser/plasma interactions cause the laser pulse shape to steepen, thereby creating strong wakefields. A witness e-beam pulse probes the wakefields. The second one, called seeded self-modulated LWFA, involves sending a seed e-beam pulse into the plasma to initiate wakefield formation. These wakefields are amplified by a laser pulse following shortly after the seed pulse. A second e-beam pulse (witness) follows the seed pulse to probe the wakefields. These LWFA experiments will also be the first ones driven by a CO(2) laser beam.

The Cape Triage Score: a new triage system South Africa. Proposal from the Cape Triage Group.

The Cape Triage Group (CTG) convened with the intention of producing a triage system for the Western Cape, and eventually South Africa. The group includes in-hospital and prehospital staff from varied backgrounds. The CTG triage protocol is termed the Cape Triage Score (CTG), and has been developed by a multi-disciplinary panel, through best available evidence and expert opinion. The CTS has been validated in several studies, and was launched across the Western Cape on 1 January 2006. The CTG would value feedback from readers of this journal, as part of the ongoing monitoring and evaluation process.

The Cape Triage Score -- a triage system for South Africa.

The Cape Triage Score (CTS) has been derived by the Cape Triage Group (CTG) for use in emergency units throughout South Africa. It can also be used in the pre-hospital setting, although it is not designed for mass casualty situations. The CTS comprises a physiologically based scoring system and a list of discriminators, designed to triage patients into one of five priority groups for medical attention. Three versions have been developed, for adults, children and infants.

Analysis of mouse brain using a clinical 1.5 T scanner and a standard small loop surface coil.

With increasing numbers of in vivo experiments in the field of neuroscience, the interest in methods for in vivo imaging of animal brains as small as those of mice has increased. Because highly specialized small bore scanners with high field strengths are not commonly available, clinical magnetic resonance imaging (cMRI) scanners have been used in the past to image rat and more recently also mouse brains in combination with specifically developed RF coils. These studies have demonstrated that imaging of small animal brains is feasible, and that tumor volumes measured by cMRI correlate well with histological tumor volume analysis. This protocol describes the cMRI settings at 1.5 T for imaging of mouse brain with resolutions up to 120 x 120 microm using an inexpensive, commercially available small loop surface coil. This allows easy establishment of a small animal MRI facility without the need for cost intensive dedicated small animal scanners or special custom made coils. In this study, we demonstrate high-resolution imaging of intracranial xenografts in a mouse glioma model and monitor the treatment effect of external field irradiation by cMRI.