A Clostridium perfringens Outbreak Traced to Improper Cooking of Prime Rib in Rochester, New York 2011

In December 2011, the Monroe County Department of Public Health investigated a report of gastrointestinal illness from two separate parties that had dined at a restaurant on the same day. An environmental and epidemiological investigation identified 17 individuals who met the outbreak case definition. A detailed questionnaire based on the restaurant’s menu was administered to patrons from both parties and statistically analyzed. Based on this investigation, it was hypothesized that consuming the prime rib (P \u3c 0.001) was associated with becoming ill. The environmental investigation indicated that the prime rib was not cooked to a proper temperature and was held at an improper temperature before being served. A prime rib sample and three stool samples from ill patrons were collected and sent for laboratory testing. Clostridium perfringens was identified in all of the samples by the New York State Department of Health (NYSDOH) Wadsworth Laboratory. This outbreak demonstrates the importance of proper food safety techniques for restaurants to prevent illness

Using Smartphone Apps in STD Interviews to Find Sexual Partners

Objectives. Smartphone applications (apps) are increasingly used to facilitate casual sexual relationships, increasing the risk of sexually transmitted diseases (STDs). In STD investigations, traditional contact elicitation methods can be enhanced with smartphone technology during field interviews. Methods. In 2013, the Monroe County Department of Public Health conducted a large, multi-infection STD investigation among men who have sex with men (MSM) using both index case and cluster interviews. When patients indicated meeting sexual partners online, disease intervention specialists (DISs) had access to smartphone apps and were able to elicit partners through access to inboxes and profiles where traditional contact information was lacking. Social network mapping was used to display the extent of the investigation and the impact of access to smartphones on the investigation. Results. A total of 14 index patient interviews and two cluster interviews were conducted; 97 individuals were identified among 117 sexual dyads. On average, eight partners were elicited per interview (range: 1–31). The seven individuals who used apps to find partners had an average of three Internet partners (range: 1–5). Thirty-six individuals either had a new STD (n=7) or were previously known to be HIV-positive (n=29). Of the 117 sexual dyads, 21 (18%) originated either online (n=8) or with a smartphone app (n=13). Of those originating online or with a smartphone app, six (29%) partners were located using the smartphone and two (10%) were notified of their exposure via a website. Three of the new STD/HIV cases were among partners who met online. Conclusion. Smartphone technology used by DISs in the field improved contact elicitation and resulted in successful partner notification and case finding

Enhanced Immunogenicity of Mitochondrial-Localized Proteins in Cancer Cells.

Epitopes derived from mutated cancer proteins elicit strong antitumor T-cell responses that correlate with clinical efficacy in a proportion of patients. However, it remains unclear whether the subcellular localization of mutated proteins influences the efficiency of T-cell priming. To address this question, we compared the immunogenicity of NY-ESO-1 and OVA localized either in the cytosol or in mitochondria. We showed that tumors expressing mitochondrial-localized NY-ESO-1 and OVA proteins elicit significantdly higher frequencies of antigen-specific CD8+ T cells in vivo. We also demonstrated that this stronger immune response is dependent on the mitochondrial location of the antigenic proteins, which contributes to their higher steady-state amount, compared with cytosolic localized proteins. Consistent with these findings, we showed that injection of mitochondria purified from B16 melanoma cells can protect mice from a challenge with B16 cells, but not with irrelevant tumors. Finally, we extended these findings to cancer patients by demonstrating the presence of T-cell responses specific for mutated mitochondrial-localized proteins. These findings highlight the utility of prioritizing epitopes derived from mitochondrial-localized mutated proteins as targets for cancer vaccination strategies.S

Detection of mammalian microRNA expression by in situ hybridization with RNA oligonucleotides

No abstract.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55962/1/21079_ftp.pd

Identification of the Rostral Migratory Stream in the Canine and Feline Brain

In the adult rodent brain, neural progenitor cells migrate from the subventricular zone of the lateral ventricle towards the olfactory bulb in a track known as the rostral migratory stream (RMS). To facilitate the study of neural progenitor cells and stem cell therapy in large animal models of CNS disease, we now report the location and characteristics of the normal canine and feline RMS. The RMS was found in Nissl-stained sagittal sections of adult canine and feline brains as a prominent, dense, continuous cellular track beginning at the base of the anterior horn of the lateral ventricle, curving around the head of the caudate nucleus and continuing laterally and ventrally to the olfactory peduncle before entering the olfactory tract and bulb. To determine if cells in the RMS were proliferating, the thymidine analog 5-bromo-2-deoxyuridine (BrdU) was administered and detected by immunostaining. BrdU-immunoreactive cells were present throughout this track. The RMS was also immunoreactive for markers of proliferating cells, progenitor cells and immature neurons (Ki-67 and doublecortin), but not for NeuN, a marker of mature neurons. Luxol fast blue and CNPase staining indicated that myelin is closely apposed to the RMS along much of its length and may provide guidance cues for the migrating cells. Identification and characterization of the RMS in canine and feline brain will facilitate studies of neural progenitor cell biology and migration in large animal models of neurologic disease

Critical Role of NADPH Oxidase in Neuronal Oxidative Damage and Microglia Activation following Traumatic Brain Injury

BACKGROUND: Oxidative stress is known to play an important role in the pathology of traumatic brain injury. Mitochondria are thought to be the major source of the damaging reactive oxygen species (ROS) following TBI. However, recent work has revealed that the membrane, via the enzyme NADPH oxidase can also generate the superoxide radical (O(2)(-)), and thereby potentially contribute to the oxidative stress following TBI. The current study thus addressed the potential role of NADPH oxidase in TBI. METHODOLOGY/PRINCIPAL FINDINGS: The results revealed that NADPH oxidase activity in the cerebral cortex and hippocampal CA1 region increases rapidly following controlled cortical impact in male mice, with an early peak at 1 h, followed by a secondary peak from 24-96 h after TBI. In situ localization using oxidized hydroethidine and the neuronal marker, NeuN, revealed that the O(2)(-) induction occurred in neurons at 1 h after TBI. Pre- or post-treatment with the NADPH oxidase inhibitor, apocynin markedly inhibited microglial activation and oxidative stress damage. Apocynin also attenuated TBI-induction of the Alzheimer's disease proteins β-amyloid and amyloid precursor protein. Finally, both pre- and post-treatment of apocynin was also shown to induce significant neuroprotection against TBI. In addition, a NOX2-specific inhibitor, gp91ds-tat was also shown to exert neuroprotection against TBI. CONCLUSIONS/SIGNIFICANCE: As a whole, the study demonstrates that NADPH oxidase activity and superoxide production exhibit a biphasic elevation in the hippocampus and cortex following TBI, which contributes significantly to the pathology of TBI via mediation of oxidative stress damage, microglial activation, and AD protein induction in the brain following TBI

Ce-Duox1/BLI-3 Generated Reactive Oxygen Species Trigger Protective SKN-1 Activity via p38 MAPK Signaling during Infection in C. elegans

Infected animals will produce reactive oxygen species (ROS) and other inflammatory molecules that help fight pathogens, but can inadvertently damage host tissue. Therefore specific responses, which protect and repair against the collateral damage caused by the immune response, are critical for successfully surviving pathogen attack. We previously demonstrated that ROS are generated during infection in the model host Caenorhabditis elegans by the dual oxidase Ce-Duox1/BLI-3. Herein, an important connection between ROS generation by Ce-Duox1/BLI-3 and upregulation of a protective transcriptional response by SKN-1 is established in the context of infection. SKN-1 is an ortholog of the mammalian Nrf transcription factors and has previously been documented to promote survival, following oxidative stress, by upregulating genes involved in the detoxification of ROS and other reactive compounds. Using qRT-PCR, transcriptional reporter fusions, and a translational fusion, SKN-1 is shown to become highly active in the C. elegans intestine upon exposure to the human bacterial pathogens, Enterococcus faecalis and Pseudomonas aeruginosa. Activation is dependent on the overall pathogenicity of the bacterium, demonstrated by a weakened response observed in attenuated mutants of these pathogens. Previous work demonstrated a role for p38 MAPK signaling both in pathogen resistance and in activating SKN-1 upon exposure to chemically induced oxidative stress. We show that NSY-1, SEK-1 and PMK-1 are also required for SKN-1 activity during infection. Evidence is also presented that the ROS produced by Ce-Duox1/BLI-3 is the source of SKN-1 activation via p38 MAPK signaling during infection. Finally, for the first time, SKN-1 activity is shown to be protective during infection; loss of skn-1 decreases resistance, whereas increasing SKN-1 activity augments resistance to pathogen. Overall, a model is presented in which ROS generation by Ce-Duox1/BLI-3 activates a protective SKN-1 response via p38 MAPK signaling

Sterile activation of invariant natural killer T cells by ER-stressed antigen presenting cells

Invariant NKT (iNKT) cells are a subset of innate-like lymphocytes that recognize lipid antigen presented on monomorphic CD1d molecules. These cells have the unique ability to shape immunity during anti-tumour immune responses and other forms of sterile and non-sterile inflammation. Recent studies have highlighted a variety of classes of endogenous and pathogen-derived lipid antigens that can trigger iNKT cell activation under sterile and non-sterile conditions. However, the context and mechanisms that drive the presentation of self-lipid antigens in sterile inflammation remain unclear. Given that ER-stress is prevalent during tumourigenesis coupled with the intersection between the ER-stress pathways and lipid metabolism, we hypothesized that ER-stressed CD1d+ cells might present immunogenic self-lipid(s) on CD1d molecules to activate iNKT cells in sterile conditions. We demonstrate that ER-stressed dendritic cells activated iNKT cells in a CD1d-dependent manner, which in turn promotes dendritic cell maturation. We illustrate that PERK signalling increases CD1d-mediated presentation of immunogenic endogenous lipid species. We demonstrate that the immunogenic self-lipid(s) are likely ceramide-based and loaded onto CD1d in the endosomal/lysosomal compartments. We also investigate the relevance of this mechanism in a human tumour setting by analysing iNKT cell responses to multiple myeloma cell lines and identifying ER-stressed myeloid cells in the tumour microenvironment. In conclusion, this work defines a previously unidentified mechanism that controls iNKT cell activation during sterile inflammation, with a potential role in human health and disease.</p

Sterile activation of invariant natural killer T cells by ER-stressed antigen presenting cells

Invariant NKT (iNKT) cells are a subset of innate-like lymphocytes that recognize lipid antigen presented on monomorphic CD1d molecules. These cells have the unique ability to shape immunity during anti-tumour immune responses and other forms of sterile and non-sterile inflammation. Recent studies have highlighted a variety of classes of endogenous and pathogen-derived lipid antigens that can trigger iNKT cell activation under sterile and non-sterile conditions. However, the context and mechanisms that drive the presentation of self-lipid antigens in sterile inflammation remain unclear. Given that ER-stress is prevalent during tumourigenesis coupled with the intersection between the ER-stress pathways and lipid metabolism, we hypothesized that ER-stressed CD1d+ cells might present immunogenic self-lipid(s) on CD1d molecules to activate iNKT cells in sterile conditions. We demonstrate that ER-stressed dendritic cells activated iNKT cells in a CD1d-dependent manner, which in turn promotes dendritic cell maturation. We illustrate that PERK signalling increases CD1d-mediated presentation of immunogenic endogenous lipid species. We demonstrate that the immunogenic self-lipid(s) are likely ceramide-based and loaded onto CD1d in the endosomal/lysosomal compartments. We also investigate the relevance of this mechanism in a human tumour setting by analysing iNKT cell responses to multiple myeloma cell lines and identifying ER-stressed myeloid cells in the tumour microenvironment. In conclusion, this work defines a previously unidentified mechanism that controls iNKT cell activation during sterile inflammation, with a potential role in human health and disease.</p

Harnessing the Power of Invariant Natural Killer T Cells in Cancer Immunotherapy

Invariant natural killer T (iNKT) cells are a distinct subset of innate-like lymphocytes bearing an invariant T-cell receptor, through which they recognize lipid antigens presented by monomorphic CD1d molecules. Upon activation, iNKT cells are capable of not only having a direct effector function but also transactivating NK cells, maturing dendritic cells, and activating B cells, through secretion of several cytokines and cognate TCR-CD1d interaction. Endowed with the ability to orchestrate an all-encompassing immune response, iNKT cells are critical in shaping immune responses against pathogens and cancer cells. In this review, we examine the critical role of iNKT cells in antitumor responses from two perspectives: (i) how iNKT cells potentiate antitumor immunity and (ii) how CD1d+ tumor cells may modulate their own expression of CD1d molecules. We further explore hypotheses to explain iNKT cell activation in the context of cancer and how the antitumor effects of iNKT cells can be exploited in different forms of cancer immunotherapy, including their role in the development of cancer vaccines