Protracted, Intermittent Outbreak of Salmonella Mbandaka Linked to a Restaurant - Michigan, 2008-2019.

In 2018, Michigan public health officials determined that a single restaurant in southwest Michigan was the source for a protracted, intermittent outbreak of Salmonella enterica serotype Mbandaka infections occurring since 2008. Isolates from 36 infected persons shared two highly related pulsed-field gel electrophoresis (PFGE) patterns and highly related whole genome sequencing (WGS) subtypes. The initial focus of the local public health investigation on food items rather than food sources (i.e., restaurants) through a questionnaire, difficulty in food history recollection among ill persons, and sporadic case identification over periods from months to years contributed to delayed source identification. The Kalamazoo County Health and Community Services Department (KHCSD) and the Michigan Department of Health and Human Services (MDHHS) collected clinical specimens, performed multiple rounds of environmental testing, and conducted multiple regulatory visits, and based on accumulated findings over 10 years, identified the restaurant source. A 2018 investigation by KCHCSD and MDHHS found that environmental samples and stool specimens from asymptomatic restaurant employees tested positive for the Salmonella Mbandaka outbreak strain. A complex association between the restaurant environment and employees resulted in patron illnesses. Environmental health interventions, facility renovation, asymptomatic employee exclusions, employee health monitoring, and recurrent facility environmental sampling measures were implemented. As a result of ongoing cases and environmental persistence of Salmonella Mbandaka, the restaurant closed permanently in 2018. Restaurant employee stool testing and environmental sampling for Salmonella early during the investigation of confirmed Salmonella cases linked to a restaurant enhances source identification. Exclusion or restriction of asymptomatic food workers with stool-positive nontyphoidal Salmonella should be considered part of restaurant outbreak mitigation.

Prior context influences motor brain areas in an auditory oddball task and prefrontal cortex multitasking modelling.

In this study, the relationship of orienting of attention, motor control and the Stimulus- (SDN) and Goal-Driven Networks (GDN) was explored through an innovative method for fMRI analysis considering all voxels in four experimental conditions: standard target (Goal; G), novel (N), neutral (Z) and noisy target (NG). First, average reaction times (RTs) for each condition were calculated. In the second-level analysis, 'distracted' participants, as indicated by slower RTs, evoked brain activations and differences in both hemispheres' neural networks for selective attention, while the participants, as a whole, demonstrated mainly left cortical and subcortical activations. A context analysis was run in the behaviourally distracted participant group contrasting the trials immediately prior to the G trials, namely one of the Z, N or NG conditions, i.e. Z.G, N.G, NG.G. Results showed different prefrontal activations dependent on prior context in the auditory modality, recruiting between 1 to 10 prefrontal areas. The higher the motor response and influence of the previous novel stimulus, the more prefrontal areas were engaged, which extends the findings of hierarchical studies of prefrontal control of attention and better explains how auditory processing interferes with movement. Also, the current study addressed how subcortical loops and models of previous motor response affected the signal processing of the novel stimulus, when this was presented laterally or simultaneously with the target. This multitasking model could enhance our understanding on how an auditory stimulus is affecting motor responses in a way that is self-induced, by taking into account prior context, as demonstrated in the standard condition and as supported by Pulvinar activations complementing visual findings. Moreover, current BCI works address some multimodal stimulus-driven systems.

eLIPS: Development and Validation of an Observational Tool for Examining Early Language in Play Settings.

Intervention in the early years can help to mitigate the risks that early language and communication difficulties pose for later learning and well-being. Critical to this is the capacity of early years educators to evaluate language development accurately in the classroom in order to target individual support effectively. This article reports on the development and testing of the Early Language in Play Settings (eLIPS) tool, an observational measure of child language. An action research model was used in the design of the tool with the result that the methodology adopted was compatible with an early years child-centered approach. Observations of children in play settings were used to gather information about early language through subscales measuring social communication, receptive and expressive language. A series of preliminary trials with 3- to 5-year-olds, established that the eLIPS measures have concurrent validity with scores from a standardized language assessment, the Clinical Evaluation of Language Fundamentals-Preschool 2 UK . Investigation of internal consistency showed reliability for use by researchers and early years educators together with inter-rater reliability across these groups. It was concluded that eLIPS has potential as a tool to assist early years educators in understanding individual patterns of language acquisition in a play-based environment and for framing team discussions about approaches to early language support.

The effects of cognitive distraction on behavioural, oculomotor and electrophysiological metrics during a driving hazard perception task.

Previous research has demonstrated that the distraction caused by holding a mobile telephone conversation is not limited to the period of the actual conversation (Haigney, 1995; Redelmeier & Tibshirani, 1997; Savage et al., 2013). In a prior study we identified potential eye movement and EEG markers of cognitive distraction during driving hazard perception. However the extent to which these markers are affected by the demands of the hazard perception task are unclear. Therefore in the current study we assessed the effects of secondary cognitive task demand on eye movement and EEG metrics separately for periods prior to, during and after the hazard was visible. We found that when no hazard was present (prior and post hazard windows), distraction resulted in changes to various elements of saccadic eye movements. However, when the target was present, distraction did not affect eye movements. We have previously found evidence that distraction resulted in an overall decrease in theta band output at occipital sites of the brain. This was interpreted as evidence that distraction results in a reduction in visual processing. The current study confirmed this by examining the effects of distraction on the lambda response component of subjects eye fixation related potentials (EFRPs). Furthermore, we demonstrated that although detections of hazards were not affected by distraction, both eye movement and EEG metrics prior to the onset of the hazard were sensitive to changes in cognitive workload. This suggests that changes to specific aspects of the saccadic eye movement system could act as unobtrusive markers of distraction even prior to a breakdown in driving performance.

Context Dependence Signature, Stimulus Properties and Stimulus Probability as Predictors of ERP Amplitude Variability.

Typically, in an oddball paradigm with two experimental conditions, the longer the time between novels the greater P3a amplitude. Here the research question is: Does an oddball paradigm maintain the greater P3a amplitude under several experimental conditions? An EEG study was carried out with an oddball number parity decision task having four conditions in control and schizophrenic participants. Contrary to previous findings (Gonsalvez and Polich, 2002; Polich, 2007) in control participants, non-correlation was found between the time of a novel (N) stimulus condition to the next novel condition and P3a amplitude. Moreover, with an innovative method for stimulus properties extraction features and EEG analysis, single trial across-subject averaging of participants' data revealed significant correlations (r > 0.3) of stimulus properties (such as probability, frequency, amplitude, and duration) on P300, and even r > 0.5 was found when N was an environmental sound in schizophrenic patients. Therefore, stimulus properties are strong markers of some of the features in the P3a wave. Finally, a context analysis of ERP waves across electrodes revealed a consistent modulation in novel appearance for MisMatch Negativity in schizophrenia. A supplementary analysis running linear modeling (LIMO) in EEG was also provided (see Supplementary Material). Therefore, in a multiple condition task: stimulus properties and their temporal properties are strong markers of some of the features in the P300 wave. An interpretation was done based on differences between controls and schizophrenics relate to differences in the operation of implicit memory for stimulus properties and stronger correlations were observed within groups related contextual and episodic processes.

The effects of array structure and secondary cognitive task demand on processes of visual search.

Many aspects of our everyday behaviour require that we search for objects. However, in real situations search is often conducted while internal and external factors compete for our attention resources. Cognitive distraction interferes with our ability to search for targets, increasing search times. Here we consider whether effects of cognitive distraction interfere differentially with three distinct phases of search: initiating search, overtly scanning through items in the display, and verifying that the object is indeed the target of search once it has been fixated. Furthermore, we consider whether strategic components of visual search that emerge when searching items organized into structured arrays are susceptible to cognitive distraction or not. We used Gilchrist & Harvey's (2006) structured and unstructured visual search paradigm with the addition of Savage, Potter, and Tatler's (2013) secondary puzzle task. Cognitive load influenced two phases of search: 1) scanning times and 2) verification times. Under high load, fixation durations were longer and re-fixations of distracters were more common. In terms of scanning strategy, we replicated Gilchrist and Harvey's (2006) findings of more systematic search for structured arrays than unstructured ones. We also found an effect of cognitive load on this aspect of search but only in structured arrays. Our findings suggest that our eyes, by default, produce an autonomous scanning pattern that is modulated but not completely eliminated by secondary cognitive load.

Induction of robust type-I CD8+ T-cell responses in WHO grade 2 low-grade glioma patients receiving peptide-based vaccines in combination with poly-ICLC.

PURPOSE: WHO grade 2 low-grade gliomas (LGG) with high risk factors for recurrence are mostly lethal despite current treatments. We conducted a phase I study to evaluate the safety and immunogenicity of subcutaneous vaccinations with synthetic peptides for glioma-associated antigen (GAA) epitopes in HLA-A2(+) adults with high-risk LGGs in the following three cohorts: (i) patients without prior progression, chemotherapy, or radiotherapy (RT); (ii) patients without prior progression or chemotherapy but with prior RT; and (iii) recurrent patients. EXPERIMENTAL DESIGN: GAAs were IL13Rα2, EphA2, WT1, and Survivin. Synthetic peptides were emulsified in Montanide-ISA-51 and given every 3 weeks for eight courses with intramuscular injections of poly-ICLC, followed by q12 week booster vaccines. RESULTS: Cohorts 1, 2, and 3 enrolled 12, 1, and 10 patients, respectively. No regimen-limiting toxicity was encountered except for one case with grade 3 fever, fatigue, and mood disturbance (cohort 1). ELISPOT assays demonstrated robust IFNγ responses against at least three of the four GAA epitopes in 10 and 4 cases of cohorts 1 and 3, respectively. Cohort 1 patients demonstrated significantly higher IFNγ responses than cohort 3 patients. Median progression-free survival (PFS) periods since the first vaccine are 17 months in cohort 1 (range, 10-47+) and 12 months in cohort 3 (range, 3-41+). The only patient with large astrocytoma in cohort 2 has been progression-free for more than 67 months since diagnosis. CONCLUSION: The current regimen is well tolerated and induces robust GAA-specific responses in WHO grade 2 glioma patients. These results warrant further evaluations of this approach. Clin Cancer Res; 21(2); 286-94. ©2014 AACR.

Antigen-specific immune responses and clinical outcome after vaccination with glioma-associated antigen peptides and polyinosinic-polycytidylic acid stabilized by lysine and carboxymethylcellulose in children with newly diagnosed malignant brainstem and nonbrainstem gliomas.

PURPOSE: Diffuse brainstem gliomas (BSGs) and other high-grade gliomas (HGGs) of childhood carry a dismal prognosis despite current treatments, and new therapies are needed. Having identified a series of glioma-associated antigens (GAAs) commonly overexpressed in pediatric gliomas, we initiated a pilot study of subcutaneous vaccinations with GAA epitope peptides in HLA-A2-positive children with newly diagnosed BSG and HGG. PATIENTS AND METHODS: GAAs were EphA2, interleukin-13 receptor alpha 2 (IL-13Rα2), and survivin, and their peptide epitopes were emulsified in Montanide-ISA-51 and given every 3 weeks with intramuscular polyinosinic-polycytidylic acid stabilized by lysine and carboxymethylcellulose for eight courses, followed by booster vaccinations every 6 weeks. Primary end points were safety and T-cell responses against vaccine-targeted GAA epitopes. Treatment response was evaluated clinically and by magnetic resonance imaging. RESULTS: Twenty-six children were enrolled, 14 with newly diagnosed BSG treated with irradiation and 12 with newly diagnosed BSG or HGG treated with irradiation and concurrent chemotherapy. No dose-limiting non-CNS toxicity was encountered. Five children had symptomatic pseudoprogression, which responded to dexamethasone and was associated with prolonged survival. Only two patients had progressive disease during the first two vaccine courses; 19 had stable disease, two had partial responses, one had a minor response, and two had prolonged disease-free status after surgery. Enzyme-linked immunosorbent spot analysis in 21 children showed positive anti-GAA immune responses in 13: to IL-13Rα2 in 10, EphA2 in 11, and survivin in three. CONCLUSION: GAA peptide vaccination in children with gliomas is generally well tolerated and has preliminary evidence of immunologic and clinical responses. Careful monitoring and management of pseudoprogression is essential.

The effect of withdrawal and intake of nicotine on smokers' ability to ignore distractors in a number parity decision task.

Nicotine's attention enhancing effects are often attributed to enhancement of stimulus filtering by the attention networks. We investigated distractibility in 20 abstinent cigarette smokers (9 hours overnight; phase 1) and tested them again after smoking one cigarette (phase 2). Their performance was compared to 20 nonsmokers (no nicotine intake). In an auditory number parity decision task, participants had to make a forced choice "odd" or "even" decision about centrally presented numbers between 2 and 9, while ignoring laterally presented preceding or simultaneous novel distractors. In phase 1, distractors that preceded goal stimuli slowed reaction times (RTs) more than simultaneously presented distractors in both groups. In phase 2, nicotine intake speeded RTs in smokers in all conditions and reduced RT variability for simple number decisions and simultaneous distractors. Overall, there was a nonsignificant trend for smokers to be less accurate than nonsmokers. Accuracy in the simultaneous distractor condition decreased in both groups in phase 2. We argue that the observed nicotine-induced improvements on behavioral performance primarily reflect enhancement of top-down control of attention.

Phase II study of induction fixed-dose rate gemcitabine and bevacizumab followed by 30 Gy radiotherapy as preoperative treatment for potentially resectable pancreatic adenocarcinoma.

BACKGROUND: Eighty percent of patients with resected pancreatic ductal carcinoma (PDC) experience treatment failure within 2 years. We hypothesized that preoperative fixed-dose rate (FDR) gemcitabine (GEM) combined with the angiogenesis inhibitor bevacizumab (BEV) and accelerated 30 Gy radiotherapy (RT) would improve outcomes among patients with potentially resectable PDC. METHODS: This phase II trial tested induction FDR GEM (1,500 mg/m(2)) plus BEV (10 mg/kg IV) every 2 weeks for three cycles followed by accelerated RT (30 Gy in 10 fractions) plus BEV directed at gross tumor volume plus a 1-2 cm vascular margin. Subjects underwent laparoscopy and resection after day 85. Therapy was considered effective if the complete pathologic response rate exceeded 10 % and the margin-negative resection rate exceeded 80%. RESULTS: Fifty-nine subjects were enrolled; 29 had potential portal vein involvement. Two grade 4 (3.4%) and 19 grade 3 toxicities (32.8%) occurred. Four subjects manifested radiographic progression, and 10 had undetected carcinomatosis. Forty-three pancreatic resections (73%) were performed, including 19 portal vein resections (44%). Margin-negative outcomes were observed in 38 (88%, 95% confidence interval [CI] 75-96), with one complete pathologic response (2.3%; 95% CI 0.1-12). There were seven (6 grade 3; 1 grade 4) wound complications (13%). Median overall survival for the entire cohort was 16.8 months (95% CI 14.9-21.3) and 19.7 months (95% CI 16.5-28.2) after resection. CONCLUSIONS: Induction therapy with FDR GEM and BEV, followed by accelerated BEV/RT to 30 Gy, was well tolerated. Although both effectiveness criteria were achieved, survival outcomes were equivalent to published regimens.

LOH in the HLA class I region at 6p21 is associated with shorter survival in newly diagnosed adult glioblastoma.

PURPOSE: Glioblastoma (GBM) shows downregulated expression of human leukocyte antigen (HLA) class I, thereby escaping from cytotoxic T cells and limiting the efficacy of immunotherapy. Loss of heterozygosity (LOH) of HLA class I (6p21) and/or ß-2 microglobulin (B2m) (15q21) regions represents irreversible downregulation. In this study, we examined the prevalence of these LOH events and their relations with overall survival in GBM. EXPERIMENTAL DESIGN: In a cross-sectional analysis on 60 adult patients with GBM, DNA from formalin-fixed, paraffin-embedded specimens were evaluated for 10 microsatellite regions of HLA class I, B2m, HLA class II, HLA class III, and 6q by PCR as well as immunohistochemical evaluation of HLA class I expression and CD8(+) T-cell infiltration. RESULTS: LOH in HLA class I, B2m, HLA class II, HLA class III, and 6q regions was present in 41.4%, 18.2%, 9.4%, 77.8%, and 36.0% of informative cases, respectively. LOH of HLA class I was associated with shorter overall survival (HR = 4.89, P = 0.0078). HLA class I was downregulated in 22% to 43% of cases based on immunohistochemistry. Cases that displayed negative staining were significantly younger. HLA class I expression correlated with intratumoral CD8(+) T-cell infiltration. CONCLUSION: LOH in the HLA class I region is frequent in adult GBMs. The association of shorter survival with LOH in this region suggests a crucial role for these genes in immunosurveillance.

MUC1 vaccine for individuals with advanced adenoma of the colon: a cancer immunoprevention feasibility study.

Cancer vaccines based on human tumor-associated antigens (TAA) have been tested in patients with advanced or recurrent cancer, in combination with or following standard therapy. Their immunogenicity and therapeutic efficacy has been difficult to properly evaluate in that setting characterized by multiple highly suppressive effects of the tumor and the standard therapy on the patient's immune system. In animal models of human cancer, vaccines administered in the prophylactic setting are most immunogenic and effectively prevent cancer development and progression. We report results of a clinical study that show that in patients without cancer but with a history of premalignant lesions (advanced colonic adenomas, precursors to colon cancer), a vaccine based on the TAA MUC1 was highly immunogenic in 17 of 39 (43.6%) of vaccinated individuals, eliciting high levels of anti-MUC1 immunoglobulin G (IgG) and long-lasting immune memory. Lack of response in 22 of 39 individuals was correlated with high levels of circulating myeloid-derived suppressor cells (MDSC) prevaccination. Vaccine-elicited MUC1-specific immune response and immune memory were not associated with significant toxicity. Our study shows that vaccines based on human TAAs are immunogenic and safe and capable of eliciting long-term memory that is important for cancer prevention. We also show that in the premalignant setting, immunosuppressive environment (e.g., high levels of MDSC) might already exist in some individuals, suggesting an even earlier premalignant stage or preselection of nonimmunosuppressed patients for prophylactic vaccination.

Pitfalls in retrospective analyses of biomarkers: a case study with metastatic melanoma patients.

BACKGROUND: Reliable prognostic biomarkers of survival and response to treatment are clearly important in oncology, and many studies have been carried out with the objective of identifying new prognostic biomarkers. Retrospective analysis of blood banked from patients is a frequently used paradigm for these studies. We describe a new study of the association of serum biomarker level with overall survival in melanoma patients, and the problems encountered in carrying it out. METHODS: Blood samples from 56 patients with stage IV metastatic melanoma were drawn prior to initiation of any treatment for their disease. Sera from the samples were stored for up to 94 months at -80°C, and were subsequently thawed at the same time and tested by multiplex Luminex assay for 30 analytes (cytokines, chemokines and growth factors). Cox regression analysis was used to assess the association between these analytes and time-to-death. RESULTS: Of the 30 analytes, 17 were associated with survival, most strongly so, and in all cases, a higher analyte level was associated with increased survival. In addition, the correlations of the levels of all possible pairs of analytes were all positive and in almost all cases highly significant. However, these results are artifacts that arise from the combination of two peculiarities of the data: the apparent decrease in analyte level with storage time, and the uniformly shorter storage times of the samples from censored patients than the storage times of the samples from patients who died. CONCLUSIONS: All retrospective studies can have hidden biases, and thus investigators should not claim new findings before examining the data in detail with the goal of determining whether the findings could be spurious. There were several suspicious findings in our initial analyses: too many analytes found significant, too many very small p-values, a uniformly positive association of analyte level with survival, and a uniformly positive correlation between analyte levels. We were convinced that these findings must be artifacts, and further analyses showed that the findings could be explained by an apparent decrease of analyte level storage time.

Phase II study of irinotecan and cetuximab given every 2 weeks as second-line therapy for advanced colorectal cancer.

BACKGROUND: Irinotecan and weekly cetuximab (I+C) is a standard second-line regimen for metastatic colorectal cancer (mCRC). This study investigated the safety and efficacy of every 2 weeks I+C in patients with mCRC. PATIENTS AND METHODS: Patients with mCRC refractory to first-line fluoropyrimidine/oxaliplatin regimens and not previously treated with I+C were eligible. Response rate (RR) was the primary endpoint. Cetuximab 500 mg/m(2) and irinotecan 180 mg/m(2) were administered intravenously (I.V.) on day 1 every 2 weeks. RESULTS: Patient characteristics (n = 31): male (n = 17), median age 62; Eastern Cooperative Oncology Group (ECOG) performance status (PS) ≤1 (n = 30), and PS = 2 (n = 1). Median number of cycles = 3 (range, 1-22). I+C doses were modified in 18 and 12 patients, respectively. Grade 3/4 adverse events: acneiform rash (n = 6); neutropenia (n = 6); and diarrhea (n = 5); there was one grade 5 respiratory failure, possibly related to therapy. Two patients had a partial response, 11 had stable disease, and 18 had progressive disease resulting in an overall RR of 6% and disease control rate of 41.9%. Median overall survival (OS) was 9.3 months (95% CI, 5.1-15), and time to progression (TTP) was 2.4 months (95% CI, 1.3-4.6). K-ras and BRAF mutations were detected in 39% and 9%, respectively, of the patients tested. There was a trend toward longer TTP among patients with wild-type K-ras and BRAF (2.6 vs. 1.7 months; P = 0.16), and OS was significantly longer in those patients (14.1 vs. 5.5 months; P = 0.04). CONCLUSIONS: The RR and TTP were lower than expected and may reflect the reduced dose intensity due to toxicities. While the OS was consistent with previous publications, the efficacy of this combination was not demonstrated.

Multiplex serum biomarker assessments: technical and biostatistical issues.

BACKGROUND: Identification of predictive and prognostic biomarkers for patients with disease and undergoing different therapeutic options is a very active area of investigation. Many of these studies seek biomarkers among circulating proteins accessed in blood. Many levels of standardization in materials and procedures have been identified which can impact the resulting data. METHODS: Here, we have observed unexpected variability in levels of commonly tested analytes in serum which were processed and stored under standardized conditions. We have identified apparent changes in cytokine, chemokine and growth factor levels detected by multiplex Luminex assay in melanoma patient and healthy donor serum samples, over storage time at -80°C. Controls included Luminex kit standards, multiplexed cytokine standards and WHO cytokine controls. Data were analyzed by Wilcoxon rank-sum testing and Spearman's test for correlations. RESULTS: The interpretation of these changes is confounded by lot-to-lot kit standard curve reagent changes made by a single manufacturer of Luminex kits. CONCLUSIONS: This study identifies previously unknown sources of variation in a commonly used biomarker assay, and suggests additional levels of controls needed for identification of true changes in circulating protein levels.

A phase I/II study of capecitabine given on a week on/week off schedule combined with bevacizumab and oxaliplatin for patients with untreated advanced colorectal cancer.

BACKGROUND: A week on/week off capecitabine schedule with oxaliplatin/bevacizumab was evaluated in patients with metastatic colorectal cancer (mCRC). PATIENTS AND METHODS: Forty patients were required. The projected median progression-free survival (PFS) was 12 months (81% power, 1-sided level 0.1 log-rank test). Capecitabine dose was 2500 mg/m(2)/day on days 1-7 (n = 11) and was increased to 3000 mg/m(2)/day (n = 29) in combination with oxaliplatin (85 mg/m(2)) and bevacizumab (5 mg/kg). Cycles were repeated every 2 weeks. RESULTS: Patient characteristics included Eastern Cooperative Oncology Group (ECOG) performance status 0 (n = 24) or 1 (n = 15); median age of 62 years (range, 38-81 years). Median cycles administered were 7 (range, 125), corresponding to 3.5 months' treatment duration. Pertinent grade 3/4 toxicities seen were diarrhea (18%), hand-foot syndrome (10%), and peripheral neuropathy (10%). Bowel perforation in 1 patient (3%) and 1 death due to a cerebral hemorrhage (3%) were noted. Response rate (RR) was 38% (1 complete and 14 partial responses). Median PFS was 8.6 months (95% confidence interval [CI], 4.7-10.2 months). Median overall survival was 17.2 months (95% CI, 10.4-24.2 months). CONCLUSION: The first US experience of capecitabine to our knowledge (3000 mg/m(2) on days 1-7) in combination with oxaliplatin/bevacizumab in mCRC does not appear to have advantages compared with current standard first-line mCRC treatment regimens.

Recommendations from the iSBTc-SITC/FDA/NCI Workshop on Immunotherapy Biomarkers.

PURPOSE: To facilitate development of innovative immunotherapy approaches, especially for treatment concepts exploiting the potential benefits of personalized therapy, there is a need to develop and validate tools to identify patients who can benefit from immunotherapy. Despite substantial effort, we do not yet know which parameters of antitumor immunity to measure and which assays are optimal for those measurements. EXPERIMENTAL DESIGN: The iSBTc-SITC (International Society for Biological Therapy of Cancer-Society for Immunotherapy of Cancer), FDA (Food and Drug Administration), and NCI (National Cancer Institute) partnered to address these issues for immunotherapy of cancer. Here, we review the major challenges, give examples of approaches and solutions, and present our recommendations. RESULTS AND CONCLUSIONS: Although specific immune parameters and assays are not yet validated, we recommend following standardized (accurate, precise, and reproducible) protocols and use of functional assays for the primary immunologic readouts of a trial; consideration of central laboratories for immune monitoring of large, multi-institutional trials; and standardized testing of several phenotypic and functional potential potency assays specific to any cellular product. When reporting results, the full QA (quality assessment)/QC (quality control) should be conducted and selected examples of truly representative raw data and assay performance characteristics should be included. Finally, to promote broader analysis of multiple aspects of immunity, and gather data on variability, we recommend that in addition to cells and serum, RNA and DNA samples be banked (under standardized conditions) for later testing. We also recommend that sufficient blood be drawn to allow for planned testing of the primary hypothesis being addressed in the trial, and that additional baseline and posttreatment blood is banked for testing novel hypotheses (or generating new hypotheses) that arise in the field.

Induction of CD8+ T-cell responses against novel glioma-associated antigen peptides and clinical activity by vaccinations with {alpha}-type 1 polarized dendritic cells and polyinosinic-polycytidylic acid stabilized by lysine and carboxymethylcellulose in patients with recurrent malignant glioma.

PURPOSE: A phase I/II trial was performed to evaluate the safety and immunogenicity of a novel vaccination with α-type 1 polarized dendritic cells (αDC1) loaded with synthetic peptides for glioma-associated antigen (GAA) epitopes and administration of polyinosinic-polycytidylic acid [poly(I:C)] stabilized by lysine and carboxymethylcellulose (poly-ICLC) in HLA-A2(+) patients with recurrent malignant gliomas. GAAs for these peptides are EphA2, interleukin (IL)-13 receptor-α2, YKL-40, and gp100. PATIENTS AND METHODS: Twenty-two patients (13 with glioblastoma multiforme [GBM], five with anaplastic astrocytoma [AA], three with anaplastic oligodendroglioma [AO], and one with anaplastic oligoastrocytoma [AOA]) received at least one vaccination, and 19 patients received at least four vaccinations at two αDC1 dose levels (1 × or 3 × 10(7)/dose) at 2-week intervals intranodally. Patients also received twice weekly intramuscular injections of 20 µg/kg poly-ICLC. Patients who demonstrated positive radiologic response or stable disease without major adverse events were allowed to receive booster vaccines. T-lymphocyte responses against GAA epitopes were assessed by enzyme-linked immunosorbent spot and HLA-tetramer assays. RESULTS: The regimen was well-tolerated. The first four vaccines induced positive immune responses against at least one of the vaccination-targeted GAAs in peripheral blood mononuclear cells in 58% of patients. Peripheral blood samples demonstrated significant upregulation of type 1 cytokines and chemokines, including interferon-α and CXCL10. Nine (four GBM, two AA, two AO, and one AOA) achieved progression-free status lasting at least 12 months. One patient with recurrent GBM demonstrated sustained complete response. IL-12 production levels by αDC1 positively correlated with time to progression. CONCLUSION: These data support safety, immunogenicity, and preliminary clinical activity of poly-ICLC-boosted αDC1-based vaccines.

Normal Gaze Cueing in Children with Autism Is Disrupted by Simultaneous Speech Utterances in "Live" Face-to-Face Interactions.

Gaze cueing was assessed in children with autism and in typically developing children, using a computer-controlled "live" face-to-face procedure. Sensitivity to gaze direction was assessed using a Posner cuing paradigm. Both static and dynamic directional gaze cues were used. Consistent with many previous studies, using photographic and cartoon faces, gaze cueing was present in children with autism and was not developmentally delayed. However, in the same children, gaze cueing was abolished when a mouth movement occurred at the same time as the gaze cue. In contrast, typical children were able to use gaze cues in all conditions. The findings indicate that gaze cueing develops successfully in some children with autism but that their attention is disrupted by speech utterances. Their ability to learn to read nonverbal emotional and intentional signals provided by the eyes may therefore be significantly impaired. This may indicate a problem with cross-modal attention control or an abnormal sensitivity to peripheral motion in general or the mouth region in particular.

A fluorinated dendrimer-based nanotechnology platform: new contrast agents for high field imaging.

OBJECTIVES: We aimed to develop a directly detected magnetic resonance imaging (MRI) contrast agent for use with high fields based on a nanoscale fluorinated dendrimer-based platform for F MRI and overcome some of the problems with F MRI. MATERIALS AND METHODS: The dendrimers were prepared in a convergent manner by making the appropriate dendron, followed by coupling to a central core. The dendrons were prepared by attaching 3 equivalents of the fluorinated amino acid to the 3 carboxylic acids of the repeat branch unit followed by deprotection of the amine branch point, and either coupling to another repeat branch unit (increasing the generation G) or used directly allowing the precise growth of the dendrimer. The size of the dendrimers was determined by diffusion nuclear magnetic resonance (NMR) spectroscopy. The toxicity of the dendrimers was measured using the MTT assay. Fluorine longitudinal relaxation time measurements were performed on a Bruker ACP-500 NMR using a saturation recovery experiment at 470.59 MHz frequency. Healthy 150 g Sprague-Dawley female rats were imaged using a dendrimer solution. RESULTS: The size of the dendrimers is generally less than 3 nm, 2 orders of magnitude smaller than the size of the perfluorocarbon nanoparticles (about 200 nm). The longitudinal relaxation time, T1, decreases with increasing dendrimer generation. A significant improvement in relaxation rate and signal-to-noise ratio can be achieved by either the chemical modification of the dendrimer with a gadolinium-chelate or by the physical addition of exogenous contrast agent. Although the dendrimers with fluorine in the surface layer are toxic, this toxicity is easily reduced by burying the fluorine further into the dendrimer interior. (19)F MR images of the rat using the dendrimer solution were rapidly obtained at 7 Tesla, the strong contrast in the heart generated by the dendrimer can be seen. CONCLUSIONS: A novel fluorinated dendrimer-based nanotechnology platform in (19)F MRI and a new bifunctional DOTA chelate were prepared and characterized. We introduce 2 methods for reducing the (19)F longitudinal relaxation time: (a) Increasing the generation; (b) covalent and noncovalent introduction of Gd(III)-chelates. A new bifunctional Gd(III)-chelate is presented. The investigations of imaging on rats suggest potential importance of the dendrimers in (19)F MRI application.