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At a rural district hospital in Burundi, a 3-year-old severely malnourished girl with Noma presented to the operating room for placement of a gastrostomy tube. The child had a large left-sided facial defect as well as trismus. After induction of general anesthesia, the anesthesia provider was unable to open her mouth. Given that the hospital has no fiberoptic bronchoscope, direct laryngoscopy and intubation were performed directly through her facial defect. After securing the airway, the surgery proceeded without difficulty.

IN the spring of 2007, a 2-year clinical drug trial began at Children’s Hospital Boston (1) with the goal to advance the quality of life for children with Hutchinson-Gilford progeria syndrome (henceforth progeria), a rare (frequency 1 in 4 million), multisystem, and inevitably fatal disease that claims young lives due to myocardial infarctions and strokes between ages 7 and 20 years (2). The journey to the first clinical trial for progeria has been facilitated by a series of collaborative scientific workshops organized by The Progeria Research Foundation (PRF) and supported by agencies interested in aging, cardiovascular disease, rare disease research, and genetics (http://www.progeriaresearch. org/2007_prf_workshop_on_progeria.html). These meetings have provided a concentrated forum to facilitate the collective thinking of clinicians and scientists about progeria, forge collaborations in this little-known field, and accelerate the discovery of new ways to push the field forward toward treatments and cure. The first PRF Scientific Workshop in 2001 helped to identify nuclear blebbing as an important phenotypic marker of progeria cells (noted by Anthony Weiss; University of Sydney, Australia), and to recognize a translocation on chromosome 1 in the cells cultured from a progeria patient (W. Ted Brown, New York State Institute for Basic Research in Developmental Disabilities), pointing geneticists in the direction of the gene mutation for progeria (3). In 2003, the second workshop was held just 3 months after publication of the gene defect in progeria, which is typically a sporadic autosomal dominant disease caused by a C ! T mutation at nucleotide 1824 of the LMNA gene encoding lamin A (4,5). Importantly, the mutation results in a persistence of an aberrant form of a farnesylated-prelamin A molecule (4) now called progerin. The attendees now included experts in the field of lamin biology and laminopathies. The 2004 Progeria Workshop, held within the National Human Genome Research Institute (NHGRI), was held specifically to discuss the role of stem cells and the potential for stem cell transplantation in progeria, an area of continuing interest for the field. At the general Progeria Workshop in 2005, two mouse models of progeria were unveiled and, although they contained identical mutations, they yielded very different phenotypes, mimicking various portions of the human disease (6,7). Early in vitro data on a potential role of farnesyltransferase inhibitors (FTIs) in treating progeria were presented by four laboratories, demonstrating that FTI could reverse nuclear blebbing in cells expressing progerin (8–11). Armed with data demonstrating that treatment of newly developed cellular assays with FTIs improves or reverses some of the effects of progerin accumulation, a clinical trial treating children with progeria with an FTI was initiated in May 2007 (1) (www.clinicaltrials.gov). The present article, detailing the 2007 PRF Scientific Workshop (November 12–14, Boston), emphasizes the complexities of progerin and lamin A biology, and the need to continue to look for the most effective protocols for correcting the effects of the lamin A defect in progeria. All speakers presented preliminary data that had not been peerreviewed at the time of presentation. In the months since the workshop, some data have been published (referenced within this article), and we look forward to additional peerreviewed publications on the work presented at this meeting over the coming year. The essence of translational research was represented in the presentations, which brought forth the most important issues in progeria today—the effects of progerin and lamins and their binding partners on the functioning of cells, systems, mouse models, and humans; the connection between progeria, aging, and cardiovascular disease in the general population; careful analysis of FTI effects on progeria at all levels (summarized in Table 1); and strategies for future disease treatments and cure. In his introduction at the workshop, Francis Collins (Director, NHGRI) noted that a 4-year period between gene discovery and a clinical drug trial is ‘‘. . . an unprecedented feat for a rare genetic disease.’’ For progeria patients and their families, who have struggled to live within a void of information, the clinical trial represents ‘‘. . . a source of light coming up after a long, long night’’ and hope has replaced ‘‘. . . a sense of fear and helplessness on each passing birthday’’ (Subbarao, father of Priya, a 15year-old with progeria).

Toll-like receptors (TLRs) regulate dendritic cell function and activate signals that mediate the nature of the adaptive immune response. The current study examined the role of TLRs in dendritic cell activation and in regulating T cell and antibody responses to antigens from the filarial parasites Onchocerca volvulus and Brugia malayi, which cause river blindness and lymphatic filariasis, respectively. Bone-marrow-derived CD11c(+) cells from C57BL/6 and TLR4(-/-) mice produced high levels of IL-6 and RANTES, and showed elevated surface CD40 expression, whereas CD11c(+) cells from myeloid differentiation factor 88(-/-) (MyD88(-/-)), TLR2(-/-) and TLR2/4(-/-) mice were not activated. Similarly, IFN-gamma production by splenocytes from immunized TLR2(-/-) mice was significantly impaired compared with splenocytes from C57BL/6 and TLR4(-/-) mice. In contrast, there was no difference among these strains in Th2-associated responses including IL-5 production by splenocytes from immunized animals, serum IgE and IgG(1), or eosinophil infiltration into the corneal stroma. Neutrophil recruitment to the cornea and CXC chemokine production was inhibited in immunized TLR2(-/-) mice compared with C57BL/6 and TLR4(-/-) mice. Taken together, these findings demonstrate an essential role for TLR2 in filaria-induced dendritic cell activation, IFN-gamma production and neutrophil migration to the cornea, but does not affect filaria-induced Th2-associated responses.

During the initial operation of the International Thermonuclear Experimental Reactor (ITER), it is envisaged that activation will be minimized by using hydrogen (H) plasmas where the reference ion ...

Over the past few years, the preparation of ITER-relevant plasma scenarios has been the main focus of experimental activity on tokamaks. The development of integrated, simultaneous, real-time controls of plasma shape, current, pressure, temperature, radiation, neutron profiles and also impurities, ELMs and MHD are now seen to be essential for further development of quasi-steady state conditions with feedback, or the stabilisation of transient phenomena with event-driven actions. For this thrust, the EFDA JET Real-Time Project has developed a set of real-time plasma measurements, experiment control and communication facilities. The plasma diagnostics used for real-time experiments are Far Infra Red interferometry, polarimetry, visible, UV and X-ray spectroscopy, LIDAR, bolometry, neutron and magnetics. Further analysis systems produce integrated results, such as temperature profiles on geometry derived from MHD equilibrium solutions. The actuators include toroidal, poloidal and divertor coils, gas and pellet fuelling, neutral beam injection, radio frequency (ICRH) waves and microwaves (LH). The heating/fuelling operators can either define a power or gas request waveform or select the real-time instantaneous power/gas request from the real-time experiment central control (RTCC) system. The real-time experiment control system provides both a high-level, control-programming environment and interlocks with the actuators. A MATLAB facility is being developed for the development of more complex controllers. The plasma measurement, controller and plant control systems communicate in ATM network. The EFDA Real-Time Project is essential groundwork for future reactors such as ITER. It involves many staff from several institutions. The facility is now frequently used in experiments. This work has been conducted under the European Fusion Development Agreement and is partly funded by Euratom and the UK Engineering and Physical Sciences Research Council.

Trace Tritium Experiments' (TTE) were successfully performed on JET in 2003. The Campaign marked the first use of tritium in JET plasmas since the Deuterium-Tritium Experiment (DTE1) Campaign in 1997, and was the first use of tritium in experiments under the EFDA organisation with the UKAEA as JET Operator. The safety and regulatory preparations for the experiment were extensive. Since JET has been operated by the UKAEA the operations have followed the model of a licensed nuclear site. The safe operation of the JET torus is demonstrated in a safety case. Key Safety Management Requirement (KSMR) and Key Safety Related Equipment (KSRE) are identified in the Safety Case for DT operation. The safe operation of the torus is within the bounds of, and under the control of, an Authority to Operate (ATO). New technical challenges were presented by the need to inject and account for small quantities of tritium in very short pulses (∼80ms), with an accurate time stamp. The safety and operational management of the campaign are described. Valuable lessons were learned which would help in running future experiments. It is concluded that JET is in a strong position to run future trace tritium and full DT discharges.

The regulatory role of cerebrospinal fluid (CSF) in brain physiology is well established, while our understanding of its role in brain immunity is undefined. We demonstrate that normal rat CSF suppresses the in vitro development of mastocytoma-specific CTL activity in restimulated splenocytes from Balb/c mice, a strain unable to reject this tumor from the brain. Suppression is dependent on TGF- β , revealed by reversal of suppression with specific neutralizing antibody. In contrast, mice which can reject this tumor from the brain, such as Balb/c mice with immunological memory to the tumor or CD-1 mice with major histo-incompatibility with the tumor, have populations of precursor CTL which are resistant to CSF-induced suppression, in the in vitro restimulation protocol. We propose that the susceptibility to CSF-induced suppression of peripherally generated immune cells that traffic to the brain plays an important role in determining whether growing tumor cells survive in the brain.

The intrathecal Ab response to Ag introduced into the normal brain has not been fully explored. Involvement of Ag-specific, peripheral B cells in an intrathecal response was studied using a normal rat model of Ag infusion through an indwelling cannula into defined brain sites, while maintaining a functionally intact blood-brain barrier. Specific Ab was detected in serum and cerebrospinal fluid. The intrathecal response is first detectable at day 14. Isoelectric focusing of cerebrospinal fluid reveals banding patterns consistent with local Ab production. To increase Ag-specific, circulating peripheral lymphocytes available for trafficking to Ag-stimulated brain and for enhancing intrathecal Ab synthesis, rats were preimmunized peripherally. Subsequently, Ag or saline (control) was infused through the cannula. Under this protocol, intrathecal synthesis is detectable earlier (day 5 postinfusion). Immunohistochemical studies at the infusion site assessed Ag-specific B cells, T cells, and activated APCs. Rats receiving peripheral preimmunization followed by Ag into caudate nucleus have far greater numbers of these cells, including plasma cells, within the infusion site compared with saline controls. Results confirm previous indirect evidence of intrathecal Ab synthesis in normal rat brain and provide the first direct evidence for B cell trafficking across normal brain barriers plus retention at the Ag deposition site. Our studies indicate that the normal brain microenvironment supports development of Ag-directed humoral immunity. We propose that immune privilege in normal brain is characterized by down-regulation of cell-mediated but not Ab immune responses within the central nervous system.

We have developed a murine model to explore the tumor-specific CTL response in the immune-privileged central nervous system using P511 mastocytoma cells. Three strains with varying degrees of histocompatibility to P511 cells (CD-1, allogeneic; BALB/c, minor histoincompatible; DBA/2, syngeneic) received tumor cells (10(4)) into the putamen 7 days after cannula implantation, when the blood-brain barrier was functionally intact. Without exception, tumor formed reproducibly by day 7 in all strains. Tumor rejection occurred in CD-1 but not in BALB/c and DBA/2 mice. Using a flank injection site, both CD-1 and BALB/c, but not DBA/2 mice, ultimately rejected flank tumors. Analysis of tumor-specific CTL in BALB/c spleens revealed that P511 administration into brain or flank elicited similar responses: no fully activated CTL were detectable but a significantly expanded population of nonkilling precursors of CTL (pCTL) were present. A P511 cell-specific pCTL population was also identified at the brain tumor site 14 days post-tumor introduction, indicating that pCTL, generated in the periphery, traffic to the tumor site in brain. These data indicate that failure to reject tumor in brain is neither due to lack of afferent stimulation nor to inability of peripheral effectors (P511 cell-specific pCTL) to reach the tumor site. We hypothesize that these effector cells are prevented from developing into fully activated CTL by conditions within the central nervous system microenvironment that down-regulate CTL development.

The JET Tokamak was fitted with four new electrodes of novel design, each powered from individual computer controlled dc and RF supplies. Details of enhancements and problems from 15 months experience with the system are outlined. Experiments were performed to assess the effect of RF on the glow discharge characteristics, and to establish stable glow at low pressure and high voltage. JET combined RF/dc glow discharge cleaning (GDC) had no significant advantages over pure dc GDC, provided highly stable dc current control was obtained. In fact, the mechanically weak electrode inductor spiral required to allow RF posed a distinct disadvantage. The electrodes were converted to simple plates, following damage caused by halo currents during Tokamak plasma disruptions. The performance of these electrodes was assessed. Future developments in the JET GDC system are outlined.

ERα-positive breast cancers comprise approximately 80% of all newly diagnosed cases. Current treatment approaches targeting ER signaling include antagonizing and/or downregulating ER or reducing estrogen levels. Faslodex (fulvestrant) is the only clinically-approved agent that is both a potent ER antagonist and downregulator but has limitations given its pharmacokinetics and route of administration. Over the past several years, targeted ER therapies have focused on developing selective estrogen receptor downregulators (SERDs, i.e, GDC-0810, GDC-0927, AZD9496, RAD1901). The mechanisms involved in ER downregulation by SERD binding are not completely understood, but evidence suggests that conformational changes in the receptor upon ligand binding combined with specific co-regulator interactions destabilize the receptor making it a target for passive proteasomal degradation. We hypothesized that the complex ER pharmacology required for SERD-based passive degradation might be different across various ER-positive cell lines and that targeted degradation of the receptor by actively recruiting the ubiquitin-proteasome machinery would provide a better approach for reducing ER levels. To test this hypothesis, we developed potent molecules directed against ER using our pioneering technology—proteolysis targeting chimeras (PROTACs). PROTACs are heterobifunctional molecules that actively recruit specific E3 ligases resulting in ubiquitylation and degradation of target proteins. When testing for ER degradation using several SERDs and ER PROTACs, we discovered that both fulvestrant and ER PROTACs provided robust degradation in all ER-positive lines (90% reduction) whereas other SERDs did not degrade or only modestly degraded the receptor. Importantly, MCF-7 cells were uniquely sensitive to SERD-based degradation of ER compared to other cell lines. Subcutaneous administration of fulvestrant (1mpk) or ER PROTACs (10 mpk) reduced uterine ER alpha levels in immature rats (>65% reduction). PROTAC-mediated degradation of ER was also achieved in breast cancer xenografts. To further validate the PROTAC mechanism, incubation of ER-positive cells with ER PROTACs resulted in increased levels of poly-ubiquitylated ERα when compared to SERDs. Lastly, to demonstrate the specificity of PROTAC-mediated ERα degradation, we utilized a cellular expression proteomics-based approach to examine over 7,000 proteins. In this experiment, only ERα and several known proteins whose genes are regulated by ERα, were significantly reduced by PROTACs. It remains to be seen how the current class of investigational downregulators will perform in the clinic. More importantly, a better understanding of the therapeutic potential and benefit of degrading the receptor instead of inhibiting the receptor needs to be explored. To that end, we continue to develop and characterize novel ER PROTACs with the anticipation that targeted ERα degradation will provide a greater clinical benefit than receptor antagonism. Citation Format: Flanagan JJ, Rossi AK, Anderoli M, Willard RR, Gordon D, Harling J, Churcher I, Smith I, Zinn N, Bantscheff M, Crews CM, Crew A, Coleman KG, Winkler JD, Qian Y. Targeted and selective degradation of estrogen receptor (ER) alpha by PROTACs [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr S4-03.

Experiments on JET with a carbon-fibre composite wall have explored the reduction of steady-state power load in an ELMy H-mode scenario at high Greenwald fraction 0.8, constant power and close to the L to H transition. This paper reports a systematic study of power load reduction due to the effect of fuelling in combination with seeding over a wide range of pedestal density ((48)×10 19m 3) with detailed documentation of divertor, pedestal and main plasma conditions, as well as a comparative study of two extrinsic impurity nitrogen and neon. It also reports the impact of steady-state power load reduction on the overall plasma behaviour, as well as possible control parameters to increase fuel purity. Conditions from attached to fully detached divertor were obtained during this study. These experiments provide reference plasmas for comparison with a future JET Be first wall and an all W divertor where the power load reduction is mandatory for operation.

Cerebrospinal fluid (CSF) and serum antibody responses to albumin administered into CSF or muscle have been compared with respect to titer, isotype profile and complement-fixing activity in a rat model with normal brain barrier function. CSF/serum titer ratios and the ratio of IgG subclasses, IgG1/IgG2, were both elevated following CSF immunization. In contrast, there was no difference in complement-fixing activity between antibodies elicited by the two routes of immunization. It is suggested that intrathecal antibody synthesis accounts for the elevated CSF antibody titers in CSF-immunized rats, providing the first example of central nervous system antibody synthesis in an animal with normal brain barrier permeability.

We have evaluated the antibody and the effector T-cell responses to a single cerebrospinal fluid (CSF) infusion of myelin basic protein (MBP) in Lewis rats by measuring serum anti-MBP antibodies and clinical signs of experimental autoimmune encephalomyelitis (EAE), respectively. Some rats developed anti-MBP antibodies, but none manifested EAE in response to the primary infusion. Antibody responses to an EAE challenge 3 weeks after CSF infusion were normal, but clinical symptoms of EAE were markedly suppressed. Brain trauma at the time of MBP pretreatment enhanced this suppression. The CSF route of MBP administration is more effective in inducing suppression of EAE than peripheral routes.

Anti-SFV antibodies generated during SFV infection may affect CNS function due to cross-reactivity with a peptide of oligodendrocyte myelin glycoprotein. To explore this possibility, total IgG from SFV immunized or normal control rabbits was unilaterally microinfused into the subthalamic region of normal rat brain. Behavior of the IgG-infused rats was determined using a bioassay, measuring rotational locomotion following systemic injection of apomorphine. Anti-SFV IgG-infused rats demonstrated a significantly increased (p0.005) ipsilateral turning response compared to control rats, persisting for at least a month. Results suggest that brain cross-reactive antibodies in anti-SFV IgG may affect brain function.

Experiments performed at JET during the past two years show that, in high triangularity H-mode plasmas with I-p = 2.5 MA, n(e)/n(Gr) approximate to 1.0, it is possible to radiate separately up to approximate to 40% of the total injected power on closed flux surfaces in the pedestal region (argon seeding) and up to approximate to 50% of the injected power in the divertor region (nitrogen seeding), while maintaining the confinement improvement factor at the value required for ITER, H98(y, 2) 1.0. The total radiated power fraction achieved in both cases (65-70%) is close to that required for ITER. However, Type I ELMS observed with impurity seeding have the same characteristics as that observed in reference pulses without seeding: decreasing plasma energy loss per ELM with increasing pedestal collisionality. One has to reach the Type III ELM regime to decrease the transient heat load to the divertor to acceptable values for ITER, although at the expense of confinement. The feasibility of an integrated scenario with Type-III ELMS, and q(95) = 2.6 to compensate for the low H factor, has been demonstrated on JET. This scenario would meet ITER requirements at 17 MA provided that the IPB98 scaling for energy content is accurate enough, and provided that a lower dilution is obtained when operating at higher absolute electron density.

In the initial phase of ITER H plasmas will be used in order to avoid activating the machine. The reference ICRF healing scenarios rely on minority species such as Helium (3He) or deuterium (D). These schemes’ distinctive feature comes from the presence of the fast magnetosonic wave ion‐ion hybrid resonance/cut‐off pair, between the antennas and the minority cyclotron layer. In order to document these unusual heating schemes, ICRF experiments were carried out recently on JET. First, the use of 3He ions in H plasmas was investigated with a sequence of discharges in which 5 MW of ICRF power was coupled to the plasma and the 3He concentration was varied from below 1% up to 10%. The inverted minority heating regime was observed at low concentrations (up to ∼2%). Energetic tails in the 3He distribution were observed with effective temperatures up to 300 keV and central electron temperatures up to 6 keV. At around 2%, a sudden transition was reproducibly observed to the mode conversion regime, in which the ICRF...

Various ITER relevant experiments using 3He in a majority D plasma were performed in the recent JET campaigns. Two types can be distinguished: dedicated studies of the various RF heating scenarios which rely on the presence of 3He, and physics studies using RF heating as a working tool to provide a tunable heat source. As the success of a number of these experiments depended on the capability to keep the 3He concentration fixed, real time control of the 3He concentration was developed and used. This paper presents a brief overview of the results obtained, zooms in on some of the more interesting recent findings and discusses some of the theoretical background.

The recent development of real-time measurements and control tools in JET has enhanced the reliability and reproducibility of the relevant ITER scenarios. Diagnostics such as charge exchange, interfero-polarimetry, electron cyclotron emission have been upgraded for real-time measurements. In addition, real-time processes like magnetic equilibrium and q profile reconstruction have been developed and applied successfully in real-time q profile control experiments using model based control techniques. Plasma operation and control against magnetohydrodynamic instabilities are also benefiting from these new systems. The experience gained at JET in the field of real-time measurement and control experiments operation constitutes a very useful basis for the future operation of ITER scenarios.

This overview of how to conduct a risk assessment highlights the use of one of the newest standards in this area ISO 14971, which covers the application of risk management, as well as ISO 10993, which covers biological evaluation of medical devices. A hypothetical risk evaluation of a device is also described.

For approximately 100 years the brain has been classified as an “immunologically privileged organ” based on the observations that tissue transplants into cerebral cortex survive longer than tissue transplants into conventional peripheral sites (Shirai 1921; Murphy and Sturm 1923; Medawar 1948; Scheinberg et al. 1966). Various conclusions and presumptions were attributed to these studies, including two predominant beliefs that: (1) there is no lymphatic drainage to alert the immune system to the presence of antigen in the brain (afferent arm of immunity), and (2) the tight endothelial junctions of the cerebral vasculature (blood-brain barrier; BBB) prevent blood lymphocytes from facilitating antigen elimination in the brain (efferent arm of immunity; for reviews see Barker and Billingham 1977; Brent 1990; Waksman 1998).

Serologic studies of children with Tourette syndrome (TS) have detected anti-neuronal antibodies but their role in TS has not been explored. Stereotypies and episodic utterances, analogous to involuntary movements seen in TS, were induced in rats by intrastriatal microinfusion of TS sera or gamma immunoglobulins (IgG) under noninflammatory conditions, as found in TS. Immunohistochemical analysis confirmed the presence of IgG selectively bound to striatal neurons. These data support the hypothesis that binding of an anti-neuronal antibody from some children with TS induced striatal dysfunction and suggest a possible cause for the basal ganglia alterations observed in children with TS.

CNS immune regulation is intimately dependent on the dynamics of cerebral extracellular fluid circulation. Animal models indicate that following the introduction of antigen into the CNS, normal circulation of interstitial and cerebrospinal fluids provides the opportunity for (a) delivery of CNS-derived antigen to lymphoid organs, as well as, (b) retention of immunologically significant amounts of antigen within the CNS. Thus, even in the absence of disease, CNS-derived antigen can induce antigen-specific activation of naive lymphocytes in lymphoid organs and specific reactivation of lymphoblasts that have migrated into the CNS. The initial peripheral immune response to CNS-derived antigen is induced in cervical lymph nodes and is characterized by a strong antibody response, no delayed-type hypersensitivity, and only priming for cytotoxic T-cell responses. This Th-2 type hierarchy of immune regulation is reinforced within the antigen-stimulated CNS where specific B lymphoblasts are permitted to develop their effector function but cell-mediated immunity is inhibited. Developing a paradigm for CNS immune regulation is important in understanding how CNS disorders in humans are induced, perpetuated, and may be manipulated.

The regulatory role of cerebrospinal fluid (CSF) in brain physiology is well established, while our understanding of its role in brain immunity is undefined. We demonstrate that normal rat CSF suppresses the in vitro development of mastocytoma-specific CTL activity in restimulated splenocytes from Balb/c mice, a strain unable to reject this tumor from the brain. Suppression is dependent on TGF-beta, revealed by reversal of suppression with specific neutralizing antibody. In contrast, mice which can reject this tumor from the brain, such as Balb/c mice with immunological memory to the tumor or CD-1 mice with major histo-incompatibility with the tumor, have populations of precursor CTL which are resistant to CSF-induced suppression, in the in vitro restimulation protocol. We propose that the susceptibility to CSF-induced suppression of peripherally generated immune cells that traffic to the brain plays an important role in determining whether growing tumor cells survive in the brain.

The intrathecal Ab response to Ag introduced into the normal brain has not been fully explored. Involvement of Ag-specific, peripheral B cells in an intrathecal response was studied using a normal rat model of Ag infusion through an indwelling cannula into defined brain sites, while maintaining a functionally intact blood-brain barrier. Specific Ab was detected in serum and cerebrospinal fluid. The intrathecal response is first detectable at day 14. Isoelectric focusing of cerebrospinal fluid reveals banding patterns consistent with local Ab production. To increase Ag-specific, circulating peripheral lymphocytes available for trafficking to Ag-stimulated brain and for enhancing intrathecal Ab synthesis, rats were preimmunized peripherally. Subsequently, Ag or saline (control) was infused through the cannula. Under this protocol, intrathecal synthesis is detectable earlier (day 5 postinfusion). Immunohistochemical studies at the infusion site assessed Ag-specific B cells, T cells, and activated APCs. Rats receiving peripheral preimmunization followed by Ag into caudate nucleus have far greater numbers of these cells, including plasma cells, within the infusion site compared with saline controls. Results confirm previous indirect evidence of intrathecal Ab synthesis in normal rat brain and provide the first direct evidence for B cell trafficking across normal brain barriers plus retention at the Ag deposition site. Our studies indicate that the normal brain microenvironment supports development of Ag-directed humoral immunity. We propose that immune privilege in normal brain is characterized by down-regulation of cell-mediated but not Ab immune responses within the central nervous system.

CFC-101, a Pseudomonas vaccine, was administered to beagle dogs by intramuscular injection for 4 weeks (5 days/week) at 0.05, 0.15 and 0.45 mg/kg/d. Clinical signs considered to be related to treatment were restricted to swelling at the injection sites, being apparent 1-2 h after treatment. There was no effect on body weight, food consumption, ophthalmoscopy, electrocardiography, hematology, biochemical and urinary parameters. The histopathological examination revealed treatment-related changes at the injection sites at all dosages, particularly in the hindlimbs where both perivascular and intramuscular aggregations of inflammatory cells were seen. Thus, the only treatment-related changes seen in this study were local reactions to the test substance at the injection sites; furthermore these changes seem to represent a pharmacological response to the test material. Because no evidence of any systemic toxicity was observed at any dosage level, it is concluded that dosages of CFC-101 up to and including 0.45 mg/kg/d were well tolerated over a period of 4 weeks in the beagle dog.

The regulatory guidelines relating to testing for immunotoxicity are not harmonised amongst the various agencies. This paper outlines the current situation within Europe, Japan and the USA. The agencies within the USA representing both agrochemicals and food additives are the most demanding in their requirements, and these guidelines are currently used as the definitive requirements for the testing of compounds for immunotoxicity.

A repeated dose toxicity study of montirelin hydrate (NS-3), a new drug for the treatment of disturbance of consciousness, was conducted in beagle dogs. The dogs were given the drug intravenously for 4 weeks at doses of 0 (control), 0.0002, 0.002, 0.02, 0.2, 2 and 20 mg/kg in males and 0, 0.2, 2 and 20 mg/kg in females. After discontinuation of the treatment, a 4-week recovery test was also conducted in the 0 and 20 mg/kg groups. No deaths related to the treatment were observed. There were no changes in body weight gain, and food and water consumptions. Nasal discharge was seen in all dose groups. Salivation, emesis and hypoactivity were observed in the 0.2 mg/kg group and over. Licking chops were seen in the 2 and 20 mg/kg groups. Trembling and agitated/restless behavior were seen in the 20 mg/kg group. Electrocardiographic examination revealed elevated heart rate in the 0.2 mg/kg group and over. Ophthalmoscopic and hematologic examinations, and urinalysis failed to show any abnormalities attributable to the treatment. Blood chemical examination disclosed increases in T3 level in the 2 and 20 mg/kg groups of males and in T4 level in the 0.2 mg/kg group and over of males. There were no pathological findings attributable to the treatment. The changes mentioned above were satisfactorily reversible. The nasal discharge seen in the 0.02 mg/kg group and below was considered to be of no toxicological significance. These results show that the NOAEL of montirelin hydrate is 0.02 mg/kg for 4-week repeated dose toxicity in dogs.

This paper reports the toxicological evaluation of pidotimod ((R)-3-[(S)-(5-oxo-2-pyrrolidinyl) carbonyl]-thiazolidine-4-carboxylic acid, PGT/1A, CAS 121808-62-6). Its acute toxicity in mice, rats and dogs was very low after oral, i.v., i.m. and i.p. administration. The repeated administration studies in rats were performed for 4 months via the i.p. route and for 12 months via the oral route. Pidotimod did not show toxic effects at dosages up to 200 mg/kg i.p. and 800 mg/kg p.o. These dosages correspond to 32.5 times the maximum dosage intended for clinical use. The repeated administration studies in dogs were performed for 26 weeks via the i.m. route and for 52 weeks via the oral route. Pidotimod did not show toxic effects at dosages up to 300 mg/kg i.m. and 600 mg/kg p.o.. It did not affect male or female rat fertility at dosages up to 600 mg/kg by oral and 500 mg/kg by i.v. route. The compound was not teratogenic in rats (600 mg/kg p.o. and 1000 mg/kg i.v.), with no effects on subsequent embryofoetal development at dosages up to 1000 mg/kg/day, and in rabbits (300 mg/kg p.o. and 500 mg/kg. i.v.). There were no peri- and postnatal toxic effects in rats (600 mg/kg p.o. and 500 mg/kg i.v.). Local tolerability of pidotimod after i.m. administration was very good. In conclusion pidotimod is characterized by a high safety margin in all animal species.

Strike-point sweeping and real-time-controlled (RTC) impurity seeding are both expected to be needed on JET following its upgrade to an all-metal wall with enhanced neutral-beam heating, thereby anticipating exhaust-control requirements plus the materials planned for ITER. Preliminary trials in the previous carbon device have combined these techniques in high-triangularity type I H-mode plasmas, using a VUV spectroscopic signal for feedback control of nitrogen injection. Compared with earlier unswept feedforward counterparts, similar strong mitigation of divertor heat load between ELMs was achieved in swept RTC cases for less than half the integrated nitrogen input and correspondingly less adverse effect upon other properties. Both sweeping and RT control contributed to this improvement. Time-average normalized energy confinement H98yt ~ 1, Greenwald density fraction fGwdt ~ 0.9 and particularly purity denoted by effective ionic charge Zefft ≈ 1.7, all remained closer to good reference levels. Transient effluxes in ELMs were also less affected, however, and would require separate active control.

The JET tokamak has been fitted with a new glow discharge cleaning system. Four electrodes of a novel design, offer the following advantages over their predecessors: they do not need water cooling and they do not take up significant port space. Results are presented for the initial use of these electrodes including current density distribution and impurity removal rate.

Cerebral extracellular fluids drain from brain to blood across the arachnoid villi and to lymph along certain cranial nerves (primarily olfactory) and spinal nerve root ganglia. Quantification of the connection to lymph in rabbit, cat and sheep, using radiolabelled albumin as a marker of flow, indicates that a minimum of 14 to 47% of protein injected into different regions of brain or cerebrospinal fluid passes through lymph. The magnitude of the outflow to lymph is at variance with the general assumption that the absence of conventional lymphatics from the brain interrupts the afferent arm of the immune response to brain antigens. The immune response to antigens (albumin or myelin basic protein) introduced into the central nervous system (CNS) has been analysed using a rat model with normal brain barrier permeability. The micro-injection of antigen into brain or cerebrospinal fluid elicits a humoral immune response, with antibody production in cervical lymph nodes and spleen, and also affects cell-mediated immunity. Furthermore, antigen may be more immunogenic when administered into the CNS than into conventional extracerebral sites. Clearly, the afferent arm of the immune response to antigens, within the CNS, is intact. Modern studies suggest that the efferent arm is also intact with passage of activated lymphocytes into the brain. Results support a new view of CNS immunology which incorporates continuous and highly regulated communication between the brain and the immune system in both health and disease.

The frequency distribution of over 600 stomach weights of deer mice showed considerable variation and was highly skewed. Because of the difficulty of defining a "full" stomach, it was not possible to use the weights of stomachs sampled to estimate consumption rates. There were no significant seasonal changes in mean stomach weights over a 3-year study and no relationship between stomach weights and sexual condition was found.

BRL 22321 is 4,9-dihydro-6,7-dimethyl-4,9-dioxo-1H-naphtho[2,3-d]-v-triazole sodium salt. It combines some of the chemical structural features of disodium cromoglycate (DSCG) and theophylline. It has mast cell stabilizing activity similar to that DSCG and in addition it has some smooth muscle relaxant activity. BRL 22321 was more potent than DSCG as an inhibitor of rat passive cutaneous and peritoneal anaphylaxis (PCA and PPA) and histamine release by antigen from passively sensitized rat peritoneal cellsin vitro. In the rat PCA test it was active when given orally and a large intravenous dose of DSCG reduced the potency of subsequent intravenous doses of either DSCG or BRL 22321, suggesting that the compounds were active by a common pathway. The compounds had similar potencies as inhibitors of the release, by antigen, of histamine and slow reacting substance of anaphylaxis (SRS-A) from passively sensitized human lung fragments. BRL 22321 was more potent than DSCG and theophylline as an inhibitor of a cyclic adenosine and a cyclic guanosine monophosphate phosphodiesterase. BRL 22321 inhibited histamine release in systems in which DSCG was inactive, for example it inhibited histamine release by antigen from chopped lung taken from hyperimmune guinea pigs and by anti-IgE from human leucocytes; in these tests it was more potent than theophylline. BRL 22321 had smooth muscle relaxant activity not shown by DSCG. It relaxed histamine elevated tone in guinea pig lung strip and tracheal spiral preparations at doses at which theophylline produced a dose dependent response. It was difficult to compare potencies since there was a difference in the nature of the responses to the drugs. DSCG had little activity over the same dose range or at higher doses. BRL 22321 also reduced increased airways resistance in the anaesthetized guinea pig produced by 5-hydroxytryptamine (5-HT) or histamine but at somewhat higher doses than those at which theophylline was active. BRL 22321 was not an antagonist of acetylcholine, histamine, 5-HT or SRS-A on the isolated guinea pig ileum.

Rats actively sensitised to bovine serum albumin (BSA) in Freund’s complete adjuvant were challenged by intraperitoneal injection of BSA. By washing out the peritoneal cavities at different times it was possible to divide the changes that occurred into two phases. The first was characterised by the release of histamine and slow reacting substance of anaphylaxis (SRS-A) with an associated extravasation of plasma proteins. The second phase was characterised by an increase in neutrophilic polymorphonuclear cells in the peritoneal cavity. Extracellular enzyme activities were also investigated. The kinetics of these events were studied and are discussed in terms of the possible mechanisms involved.