Locusta migratoria L. when exposed to two strains of Metarhizium anisopliae (Ma).Metarhizium is an important class of entomopathogenic fungi in the biocontrol of insects, but its virulence is affected by insect immunity. To clarify the mechanism in virulence of Metarhizium, we compared the immunological differences in nd and the 5th day after spray is critical in the pathogenic process.The virulence of Ma IMI330189 was significantly higher than that of Ma IBC200614 to locust, and IMI330189 overcame the hemocytes and began destroying the hemocytes of locust at 72 h after spray, while locust is immune to IBC200614. IMI330189 could overcome the humoral immunity of locust by inhibiting the activities of phenol oxidase (PO), esterases, multi-function oxidases (MFOs) and acetylcholinesterases in locust while increasing the activities of glutathione-S-transferases (GSTs), catalase and aryl-acylamidase (AA). However IBC200614 inhibit the activities of GSTs and AA in locust and increase the activities of MFOs, PO, superoxide dismutase, peroxidase and chitinase in locust. The changes of enzymes activities in period of infection showed that the time period between the 2These results found the phenomenon that Ma initiatively broke host hemocytes, revealed the correlation between the virulence of Ma and the changes of enzymes activities in host induced by Ma, and clarified the critical period in the infection of Ma. So, these results should provide guidance for the construction of efficient biocontrol Ma strains. Metarhizium anisopliae and Metarhizium acridum; therefore, the two fungi had significantly different virulence levels in locusts and cockroaches [The growing demand to reduce chemical inputs in agriculture, along with the increased resistance to insecticides, has provided great impetus for the development of alternative forms of insect-pest control. Metarhizium offers an attractive alternative to the use of chemical pesticides . Howeverkroaches . In hemokroaches , 11. Mankroaches . HoweverDuring evolution, fungi have developed mechanisms to overcome the immune systems of insects. The first mechanism involves evading the host immune system by changing their cell surface morphology and formation. For example, entomopathogenic fungi have cells with different morphologies, including no cell wall or almost no cell wall in the host hemolymph. These forms aid in the dispersion and clustering of entomopathogenic fungi in the hemocoel and can increase the surface area to absorb more nutrients. Conversely, the lower level of β-1,3-glucan in the cell surface does not stimulate the host immune response , 18. TheBeauveria bassiana spores and its secondary metabolite increase ESTs and GSTs activities in the hemolymph of infected and treated adults of Eurygaster integriceps[Multi-function oxidases (MFOs), esterases (ESTs) and glutathione-S-transferases (GSTs) are three important families of enzymes in insects that participate in the detoxification of various xenobiotics and insecticides, and play important roles in the resistance of insects to various insecticides . MFOs categriceps. Howevertegriceps, 35, andtegriceps. Thus, cMacrobrachium rosenbergii and might be increased by butane-fipronil resistance in the diamondback moth [M. anisopliae[The main role of PO in melanogenesis is to convert phenols to quinones that subsequently polymerize to form melanin . Thus, iack moth , 38. Pheack moth . After tack moth . In desenisopliae. Analysinisopliae. Howevernisopliae, and mannisopliae,45. Clar2 to H2O2, and then H2O2 is removed by POD, which needs a substrate that becomes oxidized, and CAT, which does not need a substrate that becomes oxidized [Periplaneta Americana, M. anisopliae, Isaria fumosoroseus and Hirsutella thompsonaii fungal infections initiate oxidative stress in the midgut, fat body, whole body and hemolymph, and decrease the CAT activity of fat body and midgut [Spodoptera litura, the activities of SOD, CAT, POD and specific ascorbate peroxidase in the larval body decreased after it was treated with crude destruxins at the doses that caused 90% mortality[Insects possess a suite of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD), which may form a concatenated response to the onslaught of dietary and endogenously produced oxidants , 47. SODoxidized . The actoxidized , 50. In oxidized . In Perid midgut . In Spodmortality.Thus, thM. anisopliae to Locusta migratoria L. was compared using a L. migratoria strain that had been reared for 11 generations from one oocyst in the laboratory. As observed by blood smears, the two strains of M. anisopliae, IMI330189 and IBC200614, displayed different abilities to adapt to the immune responses in the hemocytes of L. migratoria. The activities of the above enzymes in L. migratoria were compared after it was treated with IMI330189 and IBC200614 to speculate which enzymes were involved in the immune response of L. migratoria to M. anisopliae. These results can provide direction for future research to enhance the construction of toxic strains of Metarhizium.In this paper, the virulence of two fungal strains of L. migratoria were obtained from a laboratory colony that was established in 2007 from eggs collected from Hebei Province, China, and had been reared for 11 generations from one oocyst in the laboratory, without exposure to insecticides. The location  which is covered with saline-alkali soil is nearby the Bohai Sea. We have got the permission for us to conduct the field studies by Cangzhou Academy of Agriculture and Forestry Sciences of Hebei province, who is the authority department responsible for pest control in agriculture and forestry land, also with the protection of wildlife in Cangzhou. With the help of Dr. Qinglei Wang (Cangzhou Academy of Agriculture and Forestry Sciences), we collected eggs in Autumn for our laboratory experiment. This location is a natural ecosystem, it is not involving endangered or protected species during the field studies. The colony was cultured on wheat seedlings and an artificial diet in the laboratory. The nymphs were reared in a growth chamber, and then transferred to cages (60 cm long × 50 cm wide × 70 cm high) in a chamber maintained at 60 ± 5% RH, 30 ± 2°C and 14:10 (L:D).M. anisopliae strains IMI330189 was generous gifts from CABI biosciences, and IBC200614 was the serial number in our laboratory to save ATCC200614 that was purchased from ATCC (American Type Culture Collection). Two strains were selected based on their similar capabilities in penetrating the cuticle of L. migratoria when similarity of Mos1 gene for Metarhizium osmosensor-like protein of two strains was 92%, while they have different in morphology of their conidia and conidiophores and virulence to L. migratoria in previous experiment. Two funguses were cultured on potato dextrose agar (PDA) in Petri dishes at 28°C for 7–10 days under constant light, and were subcultured every 14 days. Conidia harvested from culture plates by scraping the surface of the PDA with a sterile mounted needle were collected into plastic centrifuge tubes containing 0.1% Tween 80 sterile water, and then an oscillator was used to break up any aggregates. The concentration of spores was determined using an improved Neubauer hemocytometer and adjusted to a concentration of 3.34 × 108 spores mL−1. This procedure was repeated so that a fresh suspension of spores was used for each experiment.The measure kits for measuring SOD, POD and CAT activities were purchased from Nanjing Jiancheng Bioengineering Institute . 5,5'-Dithiobis-(2-nit-robenzoic acid) (DTNB), acetylthiocholine iodide (ATChI), Catechol, Coomassie brilliant blue G-250, 1-chloro-2,4-dinitrobenzene (CDNB) and 1,2-dichloro-4-nitrobenzene (DCNB), 1-Naphthyl acetate (α-NA), 4-nitroanisole (p-NA) and NADPH were obtained from Sigma ; glutathione (GSH), Fast Blue RR salt and Bovine serum albumin were purchased from Pierce .The conidia of IMI330189 and IBC200614 were dried and preserved. The solution concentrations of the two strains were predicted by spore content and germination rate.3 to 3.34 × 107 spores mL−1. Sterile water containing 0.1% Tween 80 was used as the negative control. Third instar nymphs were identified by size and selected. A total of 45 third instar nymphs were treated with each spore concentration with potter spray tower, and three replications were performed. All of the nymphs were fed wheat seedlings. Mortality was recorded at 24-h intervals for 7 d.Five conidial suspensions in sterile water containing 0.1% Tween 80 (as wetting agent)  were pre2HPO4 + 50 mL 1/15M KH2PO4, pH 6.81) and left for 25 min. The excess liquid was removed, and slides were washed with sterile water and xylene, and then left to air dry. They were then mounted with neutral balata. The blood nucleus was dyed red or blue, while the cytoplasm was more transparent. Slides were observed and photographed under the microscope (Olympus BX61 DP72).Hemolymph was taken from the insects at 24-h intervals during infection and blood smears were made. Hemolymph was collected from the arthrodial membrane of the hindleg of the locust (control group and treated group). The membrane was first swabbed with 70% ethanol, allowed to air dry and then pierced with a sterile needle. The hemolymph was collected using a 10 μL Eppendorf Pipetman over ice to prevent coagulation. The hemolymph was dripped on slide glass, and then smeared. The smears air dried 10 min, and a drop of Giemsa dye  was added. After 2 h incubation at 56°C, 33 mL pure methanol was added. A prepared liquid of Giemsa dye: Sorensen buffer  was placed in a wet box and slides were stained for 30 min. The excess liquid was removed and slides were rinsed with tap water. They were then returned to the wet box, covered with Sorensen buffer (50 mL 1/15M Na−1 Na-phosphate buffer (PB) at pH 7 , pH 7.3  and pH 7.5 . Extracted samples were centrifuged at 10,000 ×g for 10 min at 4°C. Supernatants were transferred to new Eppendorf tubes and centrifuged at 15,000 ×g for 20 min at 4°C. Then, the supernatants were used to determine the enzymes activities and protein concentrations. 10 nymphs were tested for each enzyme.The whole bodies of third instar nymphs for each treatment were homogenized in 0.1 mol L−1)ESTs activity was assayed using the method of Han , with mo−1 CDNB and 100 μL 6 mmol L−1 GSH were added. When DCNB was the substrate, 50 μL aliquots of samples were added to microplate wells, and then 100 μL 1.2 mmol L−1 DCNB and 100 μL 6 mmol L−1 GSH were added. The reactions were measured at 340 nm every 25 s for 10 min at 27°C. Softmax Pro 6.1 was used to record and analyze the readings. For both enzymes, the enzyme activities were measured as the absorbance change rate per min (mOD min-1)The method of Oppenoorth was used with modifications . When th−1).MFOs activity was assayed using the method of Hansen and Hodgson with modifications . In each−1 DTNB and 100 μL 1.5 mmol L−1 ATChI. The reaction was measured at 405 nm every 30 s for 40 min at 27°C. Softmax Pro 6.1 was used to record and analyze the readings. The enzyme activity was measured as the absorbance change rate per min (mOD min−1)AChE activity was assayed according to the method of Han , with mo−1 catechol. The reaction was measured at 420 nm every 1 min for an hour at 27°C. Softmax Pro 6.1 was used to record and analyze the readings. The enzyme activity was measured as the absorbance change rate per min (mOD min−1)PO activity was assayed using the method of Luo , with moThe activities of CAT, SOD and POD were determined using the appropriate assay kits , according to the manufacturer’s instructions. The enzyme activities were measured as μmol/min/mg, μmol/min/mg and μmol/min/g, respectively.−1 (pH 9.1) potassium tetraborate, and the reaction was terminated by boiling at 100°C for 5 min. It was then cooled to room temperature by running water. The mixture was mixed with 1.2 mL DMAB  incubated at 37°C for 20 min, and then was cooled to room temperature by running water. The absorbance was detected at 585 nm. The enzyme activity was measured as the absorbance change rate per mg protein (mmol/min/mg).CHI activity was measured by a reducing-sugar assay. The reaction mixture (400 μL) containing 300 μL 1% (w/v) colloidal chitin and 100 μL aliquots of samples , was incAA activity was measured as described previously with minor modifications . In eachThe protein concentrations of the samples were determined by Bradford’s method  and meas50) and the slope of the concentration-mortality curve for each assay were estimated by probit analysis [L. migratoria that were treated with different concentrations of M. anisopliae. Differences among means were compared using the least significant difference test at P < 0.05.For bioassays, the median lethal concentration ; the activities of GSTs/CDNB were slightly inhibited by IMI330189, while they were increased by IBC200614; the activities of GSTs/DCNB was increased by IMI330189 with a concentration-dependent manner (P<0.05), while they were increased more by IBC200614; the activities of MFOs were inhibited by IMI330189, while they were lightly inhibited by IBC200614; the activities of AChEs were increased by IMI330189, while they were lightly inhibited by IBC200614; the activity of PO was inhibited by higher concentration of IMI330189, while it was increased by IBC200614 and lower concentration of IMI330189; the activity of SOD was increased by IMI330189 with a concentration-dependent manner(P<0.05), while it was inhibited by IBC200614; the activity of CAT was inhibited by IMI330189, while it was increased by IBC200614; the activity of POD was hardly affected by both strains; the activity of AA was increased by the lower concentration of IMI330189, while it was inhibited by IBC200614 and the higher concentration of IMI330189.From the results in L. migratoria to M. anisopliae, we measured the activities of enzymes in L. migratoria during infection by IMI330189 and IBC200614, and the results are shown in L. migratoria in the process of infection.To clarify the functions of enzymes in the immune response of From Aedes aegypti, a significant augment in the level of antimicrobial peptide (AMP) transcription correlates with infectious dose of bacteria [Insect possess a systematic and comprehensive defense system, and increased immune response are commonly associated with an increased levels of infection. In bacteria . Howeverbacteria . In our bacteria . So, we Metarhizium, but they showed a significant virulence difference to locust. Possible reasons include differences in the ability to penetrate the cuticle or to overcome hemolymph immunity and propagate in vivo. The present study found that IMI330189 and IBC200614 could penetrate the cuticle of locust and invade hemolymph. At the beginning of infection, the yeast-like cells appeared in the two treatment groups’ hemolymph, and greatly increases of their cell were found at 72 h after inoculation, while these phenomena did not appear in the control group. These results indicated that the ability of IMI330189 and IBC200614 to penetrate the host cuticle were similarly, and suggested that the difference in the virulence of IMI330189 and IBC200614 should be caused by the difference in their ability to overcome hemocytes and humoral immunity and propagate in vivo.The cellular and humoral defenses are important defense mechanism in insects against infection of pathogenic fungi in the host hemolymph, and are currently being emphasized ,66,67, bM. anisopliae, MCL1 gene functions as an anti-adhesive protective coat against phagocytosis and encapsulation, null mutant decrease the mortality of host by increasing the phagocytosis and encapsulation of host to M. anisopliae[in vivo.In overcoming host hemocytes, we found that the yeast-like cells of IMI330189 converted to hemolymph-derived hyphal bodies in locust’s hemolymph at 96 h after locust was infection by IMI330189, then circulated freely in the host hemolymph or were phagocytized by host granulocytes. These hemolymph-derived hyphal bodies could grow to a size of 3–20 μm and self-reproduction in the hemolymph and granulocytes, and could gather and initiate damage locust’s hemocytes, then killed locust when their count reached peak value at 144h after locust was infected by IMI330189. At this point, IMI330189 had completely overcome the locust’s hemocytes. However, hemolymph-derived hyphal bodies of IBC200614 were not found in locust’s hemolymph at 96 h after locust was infected by IBC200614, and IBC200614 did not kill locust at 144h after locust were infected by IBC200614. This indicated that the yeast-like cells of IBC200614 strain did not convert to hemolymph-derived hyphal bodies, and IBC200614 had not overcome the locust’s hemocytes. In nisopliae. In IMI3In overcoming humoral immunity, we found significant difference in the enzymes activities of locust at 7 days after locust was infected by IMI330189 or IBC200614. The increased PO activity can strengthen the immune system’s capability to challenge xenobiotics and healing in insects ,37,39, tst to 7th day after spray application with a single concentration of IMI330189 or IBC200614. The no obvious impact on PO activity by IMI330189 suggested that IMI330189 might overcome the immunity provided by PO, while significant increase in PO activity by IBC200614 suggested that the insects are immunized by PO. The no obvious impact on SOD and POD activities and significant increased on CAT activity by IMI330189 suggested that IMI330189 might overcome the immunity provided by antioxidant enzymes in locust by enhance the transcription of CAT, while the significant increase on SOD and POD activities and the no obvious impact on CAT activity by IBC200614 suggested that IBC200614 enhanced the transcription of SOD and POD, increasing therefore the locust’s immune response. The significant increase on GSTs/CDNB and GSTs/DCNB activities by IMI330189 suggested that IMI330189 might overcome the immunity provided by GSTs by increase the transcription of GSTs, while significant decrease on GSTs/CDNB and GSTs/DCNB activities by IBC200614 suggested that the insects are immune to IBC200614 due to the decreased of GSTs activities. The obvious decrease on ESTs activities by IMI330189 suggested that IMI330189 might overcome the immunity of ESTs by inhibit the transcription of ESTs, while the no obvious impact on ESTs activities by IBC200614 suggested that the insects are immune to IBC200614 due to the stability of ESTs activities. The significant decrease on MFOs and AChEs activities by IMI330189 suggested that IMI330189 might overcome the immunity of MFOs and AChEs by inhibit the transcription of MFOs and AChEs, while the significant increase on MFOs and AChEs activities by IBC20016 suggested that the insects are immune to IBC200614 due to the increase of MFOs and AChEs activities. The significant increase on AA activity by IMI330189 suggested that IMI330189 might overcome the immunity of locust by increase the transcription of AA, while the no obvious impact on AA by IBC200614 suggested that insects are immune to IBC200614 due to the stability of AA activities. So, IMI330189 could overcome the immune of locust by decrease the activities of PO, AChEs, ESTs and MFOs while increase the activities of CAT, AA and GSTs. However, locust could immune to IBC200614 by increase the activities of PO, MFOs, AChEs, SOD and POD while decrease the activities of GSTs and the stability of AA and ESTs activities. We speculated that the difference in above enzymes activities changes was induced by the difference in secretions between IMI330189 and IBC200614 because that the difference in above enzymes activities changes was mainly found in infection process and M. anisopliae could initiate difference variation in SOD, CAT and POD in difference insect [P. Americana, M. anisopliae fungal infections initiate oxidative stress in the midgut, fat body, whole body and hemolymph, and decrease the CAT activity of fat body and midgut [S. litura, the activities of SOD, CAT and POD in the larval body decreased after it was treated with crude destruxins at the doses that caused 90% mortality [To verify the above hypothesis, we assayed the above enzymes activities of locust from the 1e insect ,53. In Pd midgut . In S. lMetarihizium [Metarizhizium was the most important stage of virulence, the changes in enzymes activity was significantly associated to the virulence of Metarizhizium. The lower virulence strain IBC200614 significantly increased the activities of SOD, PO and MFOs on the 2nd day after spray application, and significantly increased the activities of CAT and CHI on the 3rd day after spray application. The higher virulence strain IMI330189 slightly increased the activity of PO on the 2nd day after spray application, and slightly increased the activities of SOD and POD on the 3rd day after spray application. On the 4th day after spray, IBC200614 significantly increased the activities of PO, MFOs, SOD and CAT, while IMI330189 increased the activities of PO and POD. On the 5th day after spray, IBC200614 increased the activities of PO, MFOs, SOD and CAT, and IMI330189 increased the activity of POD. So, the period from the second day to fifth day after spray of Metarizhizium was the most important stage of virulence.In previous study, the three days after spray application of the fungi has been known as the early stage of infection, and the detoxifying ESTs and GSTs may participate in the defense reactions of insect to rihizium . In thisM. anisopliae was the most important stage of virulence. These results provide insights that can be used to strengthen the virulence of M. anisopliae and aid in selecting the optimum insecticides to rotate or mix with M. anisopliae.The above results showed that the higher virulence strain IMI330189 could overcome the hemocytes and humoral immunity by decrease the activities of PO, AChEs, ESTs and MFOs and increase the activities of CAT, AA and GSTs, and the period from the second day to the fifth day after spray of S1 TableP < 0.05).Fold changes of enzymes activities were showed as means±SD. Means (±SD) followed by different lowercase letters within row of one enzyme are significantly different by Tukey’s HSD ((DOCX)Click here for additional data file.S2 TableR is an abbreviation of the correlation coefficient.(DOCX)Click here for additional data file.S3 TableR is an abbreviation of the correlation coefficient.(DOCX)Click here for additional data file.S4 TableP < 0.05).IMI is an abbreviation of IMI330189, IBC is an abbreviation of IBC200614. Fold changes of enzymes activities were showed as means±SD. Means (±SD) followed by different lowercase letters within columns of one enzyme are significantly different by Tukey’s HSD ((DOCX)Click here for additional data file.


To facilitate the implementation of the Family Smoking Prevention and Tobacco Control Act of 2009, the Federal Drug Agency (FDA) Center for Tobacco Products (CTP) has identified research priorities under the umbrella of tobacco regulatory science (TRS). As a newly integrated field, the current boundaries and landscape of TRS research are in need of definition. In this work, we conducted a bibliometric study of TRS research by applying author topic modeling (ATM) on MEDLINE citations published by currently-funded TRS principle investigators (PIs).We compared topics generated with ATM on dataset collected with TRS PIs and topics generated with ATM on dataset collected with a TRS keyword list. It is found that all those topics show a good alignment with FDA’s funding protocols. More interestingly, we can see clear interactive relationships among PIs and between PIs and topics. Based on those interactions, we can discover how diverse each PI is, how productive they are, which topics are more popular and what main components each topic involves. Temporal trend analysis of key words shows the significant evaluation in four prime TRS areas.The results show that ATM can efficiently group articles into discriminative categories without any supervision. This indicates that we may incorporate ATM into author identification systems to infer the identity of an author of articles using topics generated by the model. It can also be useful to grantees and funding administrators in suggesting potential collaborators or identifying those that share common research interests for data harmonization or other purposes. The incorporation of temporal analysis can be employed to assess the change over time in TRS as new projects are funded and the extent to which new research reflects the funding priorities of the FDA.The online version of this article (doi:10.1186/s13040-015-0043-7) contains supplementary material, which is available to authorized users. To facilitate the implementation of the Family Smoking Prevention and Tobacco Control Act (FSPTCA) of 2009, the Federal Drug Agency (FDA) Center for Tobacco Products (CTP) was formed to oversee tobacco regulatory activities. Its responsibilities include setting performance standards, reviewing premarket applications for new and modified risk tobacco products, requiring new warning labels, and establishing and enforcing advertising and promotion restrictions. In order to meet these responsibilities, the CTP has identified research priories for tobacco regulatory science (TRS) in order to inform and guide the CTP’s regulatory decision-making. While tobacco researchers have been examining some of the CTP’s TRS research priorities for many years, they have not necessarily been doing so under the umbrella or specific title of ‘tobacco regulatory science’. Therefore, examining and identifying research topics from the corpus of TRS work could help to more clearly define this growing research area. In this paper, we applied author topic modeling (ATM) , a variaLDA is known for its ability to model document contents as a mixture of topics (which comprise words describing similar things). This results in improvements in the study of hidden semantics of documents compared with previous models like Latent Semantic Indexing (LSI) , probabiHowever, modeling author-topic-word relations in TRS has not been attempted. Given the large increase in tobacco-related research, which the FDA has regulatory authority over tobacco, author topic modeling can help the field better understand the nature and scope of research already underway, and serve as a means of fostering interdisciplinary science that is needed to inform tobacco policy . MoreoveD documents is modeled as a multinomial distribution over T topics, where each topic is a multinomial distribution over W words and both sets of multinomial are sampled from a Dirichlet distribution.ATM aims at extracting information about authors and topics from a large text collection simultaneously. It is a class of Bayesian graphical model for text document collections represented by bag-of-words. In standard LDA, each document in the collection of Different from LDA, ATM incorporates authors by adding one more variable, which is uniformed assigned by a set of authors, an observed set in some corpus. As in LDA, a topic is chosen from a distribution over topics specific to that author, and the word is generated from the chosen topic.equation for author topic modeling is,α and β are Dirichlet priors for topic distributions, zid = t and yid = a are the assignments of the ith word in document d to topic t and author a respectively and xid = w indicates that the current observed word is word w. NTA represents the topic-author count matrix, where t for author a excluding the topic assignment to word wid. Similarly, NWT is the word-topic count matrix, where wth entry in the vocabulary assigned to topic t excluding the topic assignment to word wid. Finally, zid¬  and yid¬  represent the vector of topic assignments and vector of author assignment in all corpus except for the ith word of the dth document respectively.To learn the model parameters, we use Gibbs sampling where the θta, the topic distribution of each document and ϕwt, the topic distribution of each word, can be estimated with the following equations where D refers to the corpus.Following the same convention, the posterior distribution of This model can be understood as a two-stage stochastic process. An author is represented by a probability distribution over topics, and each topic is represented as probability distributions over words.http://prevention.nih.gov/tobacco/portfolio.aspx). Among the TRS PIs are 65 investigators that are part of the Tobacco Centers of Regulatory Science (TCORS), a large 14-center initiative that serves as the flagship for the TRSP. Since each article can have multiple authors, the author set considered in this work includes PIs plus the last author of the paper. The final author set includes 2,740 authors. The document set includes those MEDLINE citations with abstract available, resulting in 167,196 and 8,800 abstracts respectively. We refer to the first dataset, pulled using TRS keywords, as the KWSet and the second dataset, using publications from TRS grantees, theTRSAwardeeSet.In order to obtain a comprehensive collection of all tobacco-related research, we collected publications from two sources. The first source is 300 tobacco-related keywords developed from the FDA CTP’s key research priority and interest areas as outlined on the various TRS Funding Opportunity Annoucements’s (FOA’s), released in partnership with NIH, since the passage of the FSPTCA in 2009. The final search term list was reviewed and refined by FDA CTP and NIH ODP staff, bibliometric and tobacco research experts. The second data source is the publications from the 131 principle investigators of TRS grants funded by the CTP through the NIH’s Tobacco Regulatory Science Research Program (TRSP)  are employed for this purpose. Perplexity measures the degree how fit the topic model is to the training data while PMI measures topic coherence by calculating conformation measures of top P is the probability of wi and wj co-occur in the whole corpus and ϵ is added to avoid logarithm of zero.where cigars, smokeless tobacco, e-cigarettes and tobacco product characteristics. Therefore, in order to achieve this goal, we extracted publications, which were published from 2000 to 2013, corresponding to the four key areas from the larger KWSet. Then we divided all abstracts by year and ran author topic modeling on them respectively. Next, we calculated the proportion of key words in all the topics with K is the total number of topics (K = 400 as determined in last section), w is the key word, p(k) is the proportion of each topic and p(w|k) is the probability of the key word in Topic k.Temporal analyses of research topics can reveal interesting trends and provide guide for future endeavors. According to the FDA, there are four key areas in TRS. They are Figures We also investigated the distribution based on Mesh headings , which iAs the first step of bibliometric analysis, we made a simple count of which journals TRS researchers usually publish their articles. This step can be regarded as a compensations for author-topic modeling because we assume that the journals can associate the contexts of the articles to a large degree.The PubMed data set includes 167,196 publications from 7,134 journals. However, there are 1,824 journals from which there was only one article in the TRS keyword data set. In addition, we found that there are 5,146 journals from which there were fewer than ten articles. This indicates that it is likely these journals do not traditionally cover topics related to TRS research. Interestingly, the number of journals from which more than 100 articles were selected is 306, a much smaller and more manageable pool of potential publication outlets for TRS research. Together, those 306 journals published 155,512 of the articles in the TRS keyword data set, or 93% of all publications in the data set; so almost all TRS publications in the past have occurred in these 300 or so journals. Inspecting the top 30 journals publishing TRS articles  in the top journal list of TRS investigators, though the top journal among the two lists is the same. In addition, Addictive Behaviors is also in the top 10. From this, we can see from a different perspective what CTP funded researchers concentrate on.Top journals covered by all TRS investigators are almost identical to those of the PUBMED keyword data set  among the TCORS journal list, while it ranks 27T is determined by the grid search and comparison of perplexity defined in last section. Similar to LDA, estimation of topic distributions of words was evaluated with Log-likelihood score of the posterior distribution of words given topics, one of the standard criteria for generative model evaluation. It was found that the best perplexity for KWset was the lowest when T is 400 and the best for TRSAwardeeSet was 20 respectively. The hyperparameters α and β were fixed as 50/T and 0.01 respectively and according to the data size, the iterations for both data were 1000 and 50 respectively. The PMI evaluation for the coherence of the resulting topics yields 65% and 70% on average. These results were basically consistent with those reported for domains in news, social media or computer science. This shows that ATM can be adapted in medical fields. In order to confirm this quantitaive evaluation, more quality analysis is done in the following sections.We ran the author topic modeling developed by Steyvers and et al.  on both Figure Smoke, cigarette, cessation, abstinence, control, measure. T2, which is the second most prominent topic, contains words like intervention, health, program, network, base, train, social, prevent, address, support and community. T3 focuses on adolescent-related topics, including alcohol, family relationship and behavior. T4 is similar to T3, but emphasizing more on social elements, including school, law, industry and so on. Those topics suggest policy-making and social studies such as preventing teenager-smoking related research are one of the major trends in TRS research. Treatment is most dominant word in T5. Among them, most of words are quite relevant to this keyword. These words suggest that research on clinical practice of smoking related diseases is also tackled by TRS researchers. T6 evidently clusters research on ethnic, gender, age and surveys of smoking revealed by words American, African, white, group, population, ethnic, woman and age. In contrast, T7 talks about temporal study of smoking-related diseases since temporal words like time, year, month and clinical words like assess, measure, average, quantity, disease and datum are seen there with a good proportion.We can see that the 20 topics have comparatively balanced distributions ranging from 0.034 to 0.071. One thing worth noting is that some of the topics may be somewhat irrelevant to TRS. For those relevant to tobacco research, the topics derived have a broad diversity as discussed in the following. Top words included in the most prominent topic  from the dataset are cell is the dominant word in T8. Surrounding it are words mouse, receptor, express, airway pressure, vitro, response, inhibition, epithelial and mediation. Therefore, this topic talks more about experiments on the influence of smoking on cells. Their proportions are relatively smaller. T9 is obviously discussing relationships between smoking and cancers where cancer, risk, association, control cohort, air, lung and genotype are the prime terms. Furthermore, pollution, exposure, woman and breast also suggest that the indirect influence of smoking is included in this topic.Topics from T8 to T20 are all related to direct clinical studies because from now on, we can see there are quite a few domain-specific terms among each topic and understandably, they occupy fewer proportions due to the domain constraints. But the fewer portions do not mean that they are less important for the modeling. On the contrary, they can show how discriminative the author topic modeling can be. For example, the word child with smoking related terms asthma, screen, vaccine and HPV. As we mentioned above, all of those topics are more specialized. Without domain knowledge, it can be hard to understand why HPV is related to smoking. In fact, according to Troy et al. [Nicotine in T11 has the highest proportion, as much as 5%. It is not hard to imagine that this topic should mainly discuss nicotine and its effects. Words such as cocaine, brain, response, behavior, kg, mg, reinforce and nach prove this. T12 seems to mainly study the disorder brought by smoking and their correlations. It is composed of words including disorder, function, schizophrenium, depression, correlation, discrimination and so on. T13 comprises of a couple of rarely seen terms, such as abbreviations, DNA, NNAL -1-(3-pyridyl)-1-butanonol, which level can be affected by smoking) and nnk (4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone, one of the most prevalent and procarcinogenic compounds in tobacco), organic chemical elements, pyridyl and enzyme cancer terms, carcinogen, adduction, body and function terms, lung, liver, urinary, metabolic and so on. Among them, metabolic is the leading term unifying all of them. The majority of these topics are related to the harmful and potentially harmful constituents of tobacco products listed as one of the ten interest areas of TRS that have been highlighted by the FDA. Gene, genetic, genome sequence, individual, variant and identify in T14 show that this research topic on tobacco is from the genetic perspective while protein, mouse, regulation, binding, express and so on in T15 more from regulation and binding mechanism of the protein.As seen, the core of T10 is y et al. , a case–survival, anti, treat, apoptosis and so on. T17 seems to mainly be related to the medical absorption since we can see words like intake, concentration, ratio, oral, serum, urinary, waterpipe and others to name a few. T18 also talks about lung, but it is not about lung cancer. Instead, it is more about the general aspects of lung injury since ventilation, plasma, injury, acute and edema are there. Although smoking affects lung so much, T19 tells us that heart diseases are quite related as well where heart, cardiovascular, cardiac, vascular, endothelial, phosphoric, artery, and coronary are high frequent terms. According to Wheat et al. [T16 is also about cancer. However, different from T9, it focuses on lung cancer and treatment. The corresponding cell apoptosis can be indicated from words like Figure At the first glance, Figure Another interesting thing is to look at co-occurrence of authors among multiple topics . It can reflect two aspects, one on the closeness of two topics (the two or more can be subtopics of a big topic) and the other on interactions of two topics (they may not be related but depend on each other).genetic expressions, cell, and protein. The combination of T16 and T8 follows closely where topic 16 is about lung tumor study from gene and cell level. The topic dependence relation can be illustrated by the large number of topics co-occurring with T2 (intervention). This topic is not really funded by the FDA’s CTP, so why do they have such a high proportion of research (0.065)? If we look at other topics which investigators focus on in addition to T2, we can discover clues. Three topics occurring quite commonly with T2 are T1, T3 and T4 (4 times respectively). These four topics are about smoking cessation, vulnerable populations, and youth initiation and access, all of which are TRS priority areas. The link between smoking cessation and intervention is interesting, as interventions focusing on cessation are specifically mentioned as not a fundable TRS area. Investigators with this topic pair, which is common in tobacco control research in general, may be looking at other related topics that do fall under the TRS scope, such as nicotine reduction, consumer perception (of certain products as a cessation aid) and effective communication strategies. In addition, T7  co-occurs with T2 three times as well. This connection between temporal study and intervention would be a necessary one, as intervention research requires studies across time.It is found that T15 and T8 co-occur together 10 times, ranking the highest. This indicates that 10 authors study both topics. Both topics involve Williams D who studies 7 topics. The next four are Srivastava S (6 topics), Glantz S (5 topics), Baker T (5 topics) and Elashoff D (5 topics) respectively. Williams D, as the most diverse researcher, is in fact a leading social and behavior scientist focusing on public health [Glantz S is American Legacy Foundation Distinguished Professor of Tobacco Control at the University of California – San Francisco whose research focuses on the health effects of tobacco smoking and who is active in the nonsmokers’ rights movement and has advocated for public health polices to reduce smoking. His research topics include T1, T2, T4, T7 and T10, which quite match his research focus.If we look at authors and topics they are assigned, we see cases for two extremes in terms of involvement with a diversity of topics. Figure c health . His resBaker T is involved in T7, T10, T12, T14 and T15 while Elashoff D in topic T5, T7, T9, T12 and T16. They have two overlapping both with T7 and T12 assigned to them. Both of them seem to study topics related to treatment of smoking related diseases. What Elashoff D is studying is more cancer related. The topics both of them share are more general aspects like temporal study, function disorder and genetic tests. The remaining topics Baker T has, like T10, T14 and T15 involve smoking cessation, intervention, influences on children and protein binding and regulations. On the other hand, Elashoff D’s remaining topics including T5, T9 and T16 are all either cancer-related or organ-injury relevant. In Baker T’s webpage [Baker T concentrates on tobacco-dependence treatment and outcomes. He and his team are not only looking at smoking cessation, but also determine how quitting affects the person’s physical health, mental health, quality of life and social interactions. Then, Elashoff D’s research include statistical analysis of high-throughput microarray, biomarker discovery and validation studies. Meanwhile, he has extensive working on cancer related projects with collaborations in oral, lung, prostate, breast and skin cancers [ webpage , it stat cancers . It seemSrivastava S, a project lead on a TRS center grant, is not primarily a tobacco researcher. Instead, he is faculty in an environmental cardiology department. His topic profile includes T6, T8, T13, T15 and T16. From his webpage, we found that his research priority is toxicity, which can explain the connections of the 6 topics: all of them are less or more related to his priority. It is also a topic area prominently featured in the FDA’s TRS priority and interest areas.As mentioned before, topics discovered are not necessarily all primarily about tobacco and nicotine in this work. Instead, it focuses on finding the interactions between authors, topics and words and what trends can be traced under the frame of TRS. Observing along this thought, we found connections between tobacco and other related topics unique to TRS research. For instance, Delnevo C whose topic is T6, which is about ethnic, gender and age related study of smoking. In the website, it says that his research interests are clinical prevention services, tobacco control and survey research methods. Another one is Donny E whose topic is T11, which is about the nicotine effects. In his webpage, it says that nicotine reinforcement, regulation of tobacco and implications for healths are his primary research interests. Likewise, Farrelly M is a leading expert in tobacco control and policy interventions, for youth in particular. The only topic assigned to him is T4, exactly matching his interests.On other extreme, there are a few PIs who are only assigned one topic. One of them is metabolism, pharmacology, and legal & statistics, where the three top topic clusters are depicted in red. In the author topic network, topics are connected to authors who publications are linked to those terms. The size of the author nodes and the edges connecting them to the three topics reflect the importance of the authors and the contributions to the three topics respectively. For example, Matthay, Michael is a prominent author in the network since the node representing him has the largest size. His main research is on metabolism since the edge to it is the thickest. He is not linked to either pharmacology or legal & statistics, though, as Piccioto, Marina is ; although the latter is comparatively less prominent than the former. Both of the above authors are TCORS investigators (the blue nodes).Figure Also depicted in this figure are other TRS researchers (the green nodes). One key take-home point from Figure Above analyses are based on the pool of MedLINE abstracts of TRS PIs without distinguishing publication time and areas. For If we The trend can be seen in Figure snus and smokeless, are both steadily increasing over the time period in similar trends. While another term, snuff, decreases relatively sharply over the same time period. Chew shows a gradual decreasing trend. This shows an increased research interest over the past 10 years in the alternative smokeless tobacco product, snus.For the study of smokeless tobacco in Figure menthol is the most prominent one for this figure. Menthol shows a clear increasing ratio among the tobacco products characteristics terms, where the overall trend for all terms appears to be a decreasing one. The popularity of menthol demonstrates the increasing focus on flavorings as a research area for tobacco product characteristics [Figure eristics ,24 in reFor cigar products, the top seven key terms are displayed in Figure Electronic cigarette terms are shown in Figure In this work, we employed author topic modeling to conduct a bibliometric analysis on the publications of principle investigators on tobacco. We only reported topic interpretations and observations for the TRSAwardeeSet, as our primary interests were the TRS investigators. In fact, the KWset were diverse in both author and topics, and thus a more in-depth exploration is needed to understand this dataset. Nonetheless, we did temporal trend analysis based on the results of ATM for KWset. It showed us how the significance of key words in different topics evolved over time.Author topic modeling has been shown to be an effective approach in modeling corpus of computer sciences as well as more general ones, like publicly available emails, collections of diverse research articles. No research is done in modeling a constraint domain like tobacco regulations yet. The results show that this approach can efficiently cluster collections of articles into discriminative categories without any supervision. More interestingly, it can associate topics to authors in a high accuracy. This indicates that we may incorporate author topic modeling into author identification systems to infer the identity of an author of articles using topics generated by the model.The relevance of this analysis to TRS is multiple folds. First, this analysis is a ‘proof of concept’ that it can be beneficial to assess the change over time in TRS as new projects are funded and collaborative science in this area changes. This is particularly important because the FDA must use the data from funded research to inform their regulatory decision-making, so if there are ‘holes’ in the types of research being conducted or published, a bibliometric analysis with ATM could be helpful for the FDA to make decisions. The results can be used to assess the extent to which new research reflects the funding priorities of the FDA.Second, ATM outcomes can be used by investigators to assess who is conducting research in a particular research domain in order to foster collaborative science ,26. AgaiThird, these analyses begin to demonstrate the evolving research productivity of investigators, which we anticipate will occur to a greater extent as publications increase due to FDA funding. For example, we found that ‘cessation’ and ‘treatment’ clustered even though that topic is not really included in tobacco regulatory science. This clustering seems to reflect that some leading scientists who conducted research on tobacco treatment have successfully either shifted or expanded their research focus on tobacco regulatory science. Future analyses can further delineate how scientists transition into tobacco regulatory science research, particularly as a result of new funding, to better understand both the scientific expertise relevant to the TRS field, and also to understand the impact in other fields of scientists following the increased funding in TRS.By fostering collaborative science in TRS, it becomes possible to speed advances in that science by fostering communication between scientists that can avoiding un-needed duplication and impact decision-making on new science that can benefit regulatory decision-making.One limitation for this approach is that author topic modeling assumes that the topic distribution of each word in one document is only associated with one of the known authors. As a result, correlations of authors cannot be reflected from words of the same document and instead, must be found across multiple documents, which have the same authors. For large amount of corpus, this may not be a big problem. Nonetheless, this limitation can be overcome if we introduce the topic-author as multiple to multiple. Namely, instead of sampling one author each time, we allow sampling more than one. Then, the topic distribution will be generated by the joint distribution of more than one author. This way, each word will be associated with more than one author and thus, a multiple to multiple word-author interactions will be constructed. This will lead to more complicated inference algorithms. More high efficient optimization algorithms are thus needed in our future work.The other limitation of our work is the one to one author-word correspondence. Hence, in our future study, we will extend author topic modeling into group author topic modeling. In addition, considering that research topics may change every few years even for the same investigators, it would therefore be reasonable to model temporal changes. One more extension can be that we may build a predictive model based on author topic modeling so that we can assign authors to unknown articles or we can predict what main topics an unknown article is about. Yet another limitation is the lag between publication date and current research activity. Given the rapidly changing nature of research and funding in the area of tobacco regulatory science, it is very possible that investigators have moved into different research domains relative to their publication record. This is particularly relevant in the tobacco regulatory science area because it is a relatively new research domain that has caused some scientists to shift their research focus in order to obtain funding that is specifically relevant to the needs of the FDA. Thus, data from the author topic modeling could provide a misleading perspective on current research activities of scientists.Besides addressing those limitations, we plan to experiment author topic modeling with domain-specific ontologies or information models instead of only the bag of words. One such ontology is the MeSH indexing widely used in PubMed MedLINE. For articles indexed by PubMed, usually about 10 MeSH terms will be assigned to them so that the reader can easily find the theme. Therefore, those MeSH terms can be utilized as key words in doing author topic modeling or alternatively, MeSH terms can be employed as the author variable so that we can construct a mapping from MeSH term to texts. Beyond, since we have collected a lot of data and explored concepts and relations with the help of ATM, it is possible that we may build our own domain ontology independently from MeSH Indexing for tobacco related research and then align or merge with MeSH indexing for future research on data mining.Another possible extension is that we will attempt to access full texts rather than only abstracts and meanwhile, construct citation links from the reference section. Enriched by full text and citaiton links, we believe that the correlations of research topics in tobacco regular science can be more fully revealed.


Vibrio vulnificus. However, cases of severe septicemia, necrotizing fasciitis, and peritonitis caused by V. vulnificus in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) have not been described. We report a case of severe septicemia, necrotizing fasciitis, and peritonitis due to V. vulnificus in a patient undergoing CAPD after ingesting raw seafood.Chronic kidney disease, including end-stage renal disease, has been identified as a possible risk factor for primary septicemia and wound infection by V. vulnificus infection. Finally, V. vulnificus was confirmed by 16S ribosomal ribonucleic acid gene sequencing using blood and peritoneal effluent fluid cultures. The administration of intraperitoneal ceftazidime and intravenous ciprofloxacin/ceftriaxone was continued for 4 weeks, and the patient completely recovered.A 37-year-old woman undergoing CAPD was admitted to the emergency room due to general weakness, fever, diarrhea, and abdominal pain. Although empirical intraperitoneal antibiotics were administered for the diagnosis of CAPD-related peritonitis, her fever did not subside. On hospital day 3, she had hemorrhagic bullae on both lower legs. We evaluated her recent food history, and found that she ate raw seafood before admission. She underwent emergency fasciotomy on the suspicion of necrotizing fasciitis by V. vulnificus infection in vulnerable patients who ingest raw seafood is essential for prompt diagnosis, which could significantly improve patient outcomes.Suspicion of  Vibrio vulnificus is a species of gram-negative, motile, curved, rod-shaped pathogenic bacteria recovered from estuary, marine, and brackish environments [Vibrio species pathogenic to humans in 1976 [V. vulnificus infection is one of the few foodborne illnesses with an increasing incidence [V. vulnificus infections are gastrointestinal illness, primary septicemia, and wound infections. Even with prompt diagnosis and aggressive therapy, the severity of infection is high, with >90 % of infected patients requiring hospitalization and a case-fatality rate of 30–50 % [ronments . It was  in 1976 . V. vulnncidence . The maiV. vulnificus are immunocompromised, and the common underlying conditions are alcoholic liver disease, hepatitis B or hepatitis C or viral-related cirrhosis, diabetes mellitus, and steroid use [V. vulnificus infection. Renal disease might account for up to 7–12 % of primary septicemia cases [V. vulnificus peritonitis after consuming or handling raw seafood in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) [V. vulnificus in patients undergoing CAPD have not been described. Here, we report a case of severe septicemia, necrotizing fasciitis, and peritonitis due to V. vulnificus, which was identified by 16S ribosomal ribonucleic acid (rRNA) gene sequencing, in a patient undergoing CAPD after ingesting raw seafood. Informed consent was obtained from the patient and the study protocol was approved by the Institutional Review Board of Chonnam National University Hospital, South Korea (CNUH-EXP-2015-129). This study was conducted according to the principles of the Declaration of Helsinki.Almost all patients with sepsis due to roid use . Pre-exiia cases . There hs (CAPD) . However9/L (4.8–10.8 × 109/L), hemoglobin level of 8.6 g/dL (12–18 g/dL), impaired liver function as shown by aspartate and alanine transaminase levels of 48 U/L and 16 U/L, respectively (10–37 U/L), C-reactive protein level of 17.5 mg/dL (0–0.3 mg/dL), procalcitonin level of 4.23 ng/mL (0–0.5 ng/mL), iron level of 50 μg/dL (29–164 μg/dL), transferrin saturation of 43.4 % (25–50 %), and ferritin level of 3207 μg/L (4.6–274 μg/L). The peritoneal effluent fluid was cloudy with a WBC count of 560 × 106/L and 55 % polymorphonuclear leukocytes. After performing peritoneal fluid and blood cultures, the patient was treated empirically with intraperitoneal cefazolin (600 mg) and ceftazidime (1 g) once daily. Two days after initiating antibiotic therapy, the abdominal pain subsided and peritoneal effluent fluid was cleared. However, her body temperature remained at 39.1 °C. On hospital day 3, she had severe painful leg edema abruptly. Moreover, she had cellulitis with ecchymosis and progressive hemorrhagic bullae on both lower legs. Shortly after, her blood pressure decreased to 60/40 mmHg. We could conduct any radiological investigations due to cardiovascular instability. She was then admitted to the intensive care unit. We briefly evaluated her recent food history, and found that she consumed salted baby octopuses and undercooked cockles few days before admission. On the suspicion of necrotizing fasciitis by V. vulnificus infection, she underwent emergency bedside fasciotomy. Wound cultures were performed during the fasciotomy, but showed no bacteria growth. On hospital day 4, peritoneal effluent fluid and blood cultures performed at admission revealed V. vulnificus. Therefore, we decided to perform a more accurate identification using 16S rRNA gene sequencing.A 37-year-old woman with end-stage renal disease (ESRD) secondary to systemic lupus nephritis was admitted to the emergency room due to general weakness, fever, diarrhea, and abdominal pain. She had been undergoing CAPD for 15 years. On physical examination, she was found to have abdominal and rebound tenderness, but no skin wound. On admission, her body temperature was 38.8 °C, pulse was 101 beats/min, and blood pressure was 80/40 mmHg. Redness or discharge at the exit site of the peritoneal catheter was not observed. The patient had the following laboratory findings (reference ranges are indicated within parentheses): white blood cell (WBC) count of 5.3 × 10V. vulnificus (GenBank accession number: KP223908.1), which was confirmed with nucleotide basic local alignment search tool analysis. V. vulnificus showed sensitivity to cefotaxime, ciprofloxacin, and ceftazidime, intermediate sensitivity to cefazolin, and resistance to cefoxitin using the broth microdilution methods. Therefore, intraperitoneal cefazolin was discontinued, but intravenous ciprofloxacin (200 mg twice a day) and cefotaxime (1 g every 24 h) were administered in addition to intraperitoneal ceftazidime (1 g once daily) after dose adjustment for renal insufficiency. After 7 days of treatment, the peritoneal effluent WBC count decreased to 3 × 106/L. Furthermore, the absence of bacterial growth was confirmed with follow-up blood and peritoneal fluid cultures. The intraperitoneal and intravenous antibiotics were continued for 4 weeks. The patient underwent additional debridement for granulation tissues in both lower legs after emergency fasciotomy, and her wound completely healed after 9 months. The patient had been advised to avoid consumption of raw and uncooked seafood, and use separate cutting boards and knives for seafood and non-seafood. Furthermore, she was instructed to wear gloves when handling raw oysters or shellfish.Genomic deoxyribonucleic acid was extracted from the cultivated microorganism using proteinase K. The 16S rRNA gene was successfully amplified with universal primers  using Biometra T3000  and sequenced using Applied Biosystems 3130 lx . The 16S rRNA gene sequences were 100 % homologous to V. vulnificus is an opportunistic human pathogen transmitted from seawater, raw oyster, and shellfish. The spectrum of disease can vary from gastroenteritis to severe septicemia and necrotizing fasciitis.V. vulnificus infections. Patients with alcoholic liver disease, diabetes mellitus, gastrointestinal and hematologic disorder, malignancy, and iron overload states have a higher risk of infection [V. vulnificus infection. Functional abnormalities of neutrophils, monocytes, dendritic cells, and lymphocytes have been observed in uremia, which makes patients prone to specific life-threatening infections [V. vulnificus infection by enhancing the growth of the organism and possibly reducing the activity of neutrophils [V. vulnificus grows rapidly when transferrin saturation exceeds 70 % [Patients with certain underlying medical conditions are susceptible to nfection . Moreovefections . In additrophils . V. vulneds 70 % . In thisV. vulnificus infection has not been well described in patients undergoing peritoneal dialysis. Interestingly, there are only 2 case reports of V. vulnificus peritonitis after consuming or handling seafood in patients undergoing CAPD, and these patients completely recovered after oral and intraperitoneal antibiotic treatments [V. vulnificus infections in Korea and the United States has rapidly increased during the last decade due to the increased consumption of raw fish or shellfish [V. vulnificus peritonitis, including our report, were conducted in Asia, patients undergoing CAPD and live in an area with high incidence of V. vulnificus infection should avoid raw or undercooked seafood to prevent infection. A remarkable finding about our case was that peritonitis was followed by severe septicemia and necrotizing fasciitis. We could presume that V. vulnificus in the peritoneal fluid might penetrate the peritoneum or bowel wall into the blood stream, which results in septicemia. Therefore, peritonitis might be an important clinical manifestation of V. vulnificus infection in peritoneal dialysis patients. Furthermore, according to our findings, patients undergoing peritoneal dialysis are highly susceptible to various degrees of V. vulnificus infection from peritonitis to septicemia and necrotizing fasciitis.eatments , 8. Simihellfish , 3. AlthVibrio species. However, Vibrio and other closely related species show similar phenotypic features and are not easily distinguished biochemically [Vibrios isolated from the aquaculture environment has been imprecise and requires many biochemical and/or physiological tests. Although the 16S rRNA gene sequencing method may be less reliable because many different species within the genus Vibrio may contain identical 16S rRNA gene sequences, it remains one of the most commonly used methods for bacterial identification [V. vulnificus that are difficult to identify using phenotypic and biochemical methods.An array of phenotypic and genomic techniques has become available for the identification of emically . The idefication . 16S rRNV. vulnificus is tetracycline family  plus third-generation cephalosporin [V. vulnificus [V. vulnificus infection.Moreover, the recommended antibiotic therapy for losporin . Alternalosporin . In a relnificus . TherefoV. vulnificus infection, our patient recovered without complication. Necrotizing fasciitis is a life threatening soft tissue infection with a high mortality rate, which requires emergent surgical fasciotomy, debridement, and broad-spectrum antibiotic treatment when the diagnosis is confirmed [V. vulnificus infection in vulnerable patients who ingest raw seafood is essential for prompt diagnosis, which could significantly improve patient outcomes.Despite reports of high mortality from onfirmed . ConsequWritten informed consent for publication of this case and any accompanying images was obtained from the patient. A copy of the written consent is available for review by the Editor of this journal.


We propose a novel local nearest neighbor distance (LNND) descriptor for anomaly detection in crowded scenes. Comparing with the commonly used low-level feature descriptors in previous works, LNND descriptor has two major advantages. First, LNND descriptor efficiently incorporates spatial and temporal contextual information around the video event that is important for detecting anomalous interaction among multiple events, while most existing feature descriptors only contain the information of single event. Second, LNND descriptor is a compact representation and its dimensionality is typically much lower than the low-level feature descriptor. Therefore, not only the computation time and storage requirement can be accordingly saved by using LNND descriptor for the anomaly detection method with offline training fashion, but also the negative aspects caused by using high-dimensional feature descriptor can be avoided. We validate the effectiveness of LNND descriptor by conducting extensive experiments on different benchmark datasets. Experimental results show the promising performance of LNND-based method against the state-of-the-art methods. It is worthwhile to notice that the LNND-based approach requires less intermediate processing steps without any subsequent processing such as smoothing but achieves comparable event better performance. Due to the fact that anomaly is a potentially hazardous source in crowded scenes, anomaly detection in video surveillance is an important task for public security and safety and attracts more and more researchers' attentions recently. It is also a challenge task, because it requires inspecting an excessive number of pedestrians or moving objects and their activities and overcomes some difficult problems such as frequent occlusions, illumination change, noisy, and deformation. The primary task of anomaly detection is to detect the event or interaction that deviated from the expected .  FigureAnomalies can be classified into single anomalous event and anomalous interaction involving multiple events. Single anomalous event is defined by native information of the event such as anomalous speed, direction, and appearance. Anomalous interaction is defined by spatial context around the event with respect to another events occurring at the same time. There is no doubt that the interaction between anomalous event and the other event is anomalous, even between two normal events may be anomalous. For example, one man appears in front of running car. In most previous works, video event is typically defined by native information within a predefined spatiotemporal region and characterized by low-level feature descriptors. The commonly used low-level feature descriptors include optical flow-based –11, gradIn video scenes analysis, contextual information refers to the spatiotemporal relationships between it and other events or its located surroundings , which cDue to the richer information content contained in the high-dimensional feature descriptor, video events are typically characterized by high-dimensional feature descriptor for adapting its diversity and complexity. In , 13–17, In this paper, we propose a novel LNND descriptor to represent video event for detecting abnormal event in crowded scenes. First, the contextual information is incorporated into LNND descriptor, so both anomalous single event and anomalous interaction of multiple events can be detected without additional learning contextual information by spatiotemporal model. Second, LNND is a concise and compact descriptor that its dimensionality is much lower than that of low-level feature descriptor, because it is constructed by considering only a few spatial and temporal neighbor events. Accordingly, the memory requirement for storage and computation time for training detector can be saved. In order to tackle feature's noise and uncertainty which is inevitable in crowded scenes, EMD  is adoptThe main contributions of our work are as follows. (1) We propose a simple yet efficient LNND descriptor to represent video event for anomaly detection in crowded scenes. By using LNND descriptor, contextual information can be accounted for, so both anomalous event and interaction can be efficiently detected with less intermediate process. (2) Due to the fact that dimensionality of LNND descriptor is much lower than typically used low-level feature descriptor, both memory requirement and computation time can be accordingly saved. To our best knowledge, this is the first attempt to represent the video event by local nearest neighbors distance. We use the very concise descriptor and yet achieve the performance that can be comparable with the state-of-the-art methods; therefore, our idea is in accordance with the rule of Occam's razor .The rest of this paper is organized as follows. Anomaly detection in video surveillance is a hot topic and attracts more and more researchers' interest. Meanwhile, there is a challenge task due to many difficult problems, such as inevitable noise illumination change and deformation in the scenes, diversity of event, and interaction between multiple events. To detect anomaly in crowded scenes, different methods have been proposed to overcome one or more specific problems. These methods can be categorized into two classes according to the used feature descriptor: one is the tracking-based methods and the other is the nontracking-based method.For tracking-based methods –31, the To avoid the tracking problem in crowded scenes, many nontracking-based methods have been proposed. In these methods, the used feature descriptors, such as optical flow, gradient, and texture-based feature descriptors, were extracted from local 2D region, 3D clip, or local cuboids , 32–34. V, we divide it into a set of spatiotemporal cuboids  {Vs,t}, where s and t are the locations of the cuboid in spatial and temporal, respectively. Each Vs,t is considered as an event, and all of them have uniform size of h × w × τ and partially overlapped with its neighbor cuboids. Let Xs,t denote the low-level feature descriptor extracted from Vs,t. We compute the distance between the local event Vs,t and each of its neighbors Vs′,t′; that is, compute d. In order to cope with the inevitable noise and uncertainty in the low-level feature descriptor, we adopt EMD as distance measure. Next, we start by introducing the low-level feature extraction.In this section, we describe the detail of the construction of LNND descriptor. Given a video sequence V ∈ Rh×w×τ obtained by dividing video sequence, we compute spatiotemporal gradient magnitude for each pixel. Then, the 2nd , 3rd , and 4th  central moments are computed for each spatial pixel location along the temporal direction, which reflect three important statistical properties, that is, variance, skewness, and kurtosis, of the temporal change of the pixel spatiotemporal gradient magnitude, respectively. We define the moment matrix Mr, r = {2,3, 4} associated with V as follows:vijt is the spatiotemporal gradient magnitude value of the pixel at location {i, j} of the tth patch. Each moment matrix Mr, r = {2,3, 4} is transformed to a vector mr ∈ Rhw. The three moment vectors corresponding to three values of r are concatenated on top of each other to form a single vector m ∈ RD, where D = 3hw:M is the computed LMP descriptor. After extracting the low-level feature descriptor for each event, we will present the distance measure between two events in the next subsection.Generally speaking, most pervious used low-level features can be served for our purpose, such as multiscale histogram of optical flow (MHOF) , 9, 11, p, q ∈ RD be two histograms and are all normalized to unit mass. The EMD is obtained as the solution of the transportation problem:fi,j denotes the flow between bi and qi and gi,j denotes the ground distance between i and j. This problem can be solved by considering it as a linear dynamic programming problem. However, in the case of high-dimensional histograms, solving . Many efforts had been devoted to reduce the complexity and speed up the calculation of EMD. In 1Q, [c]1C}.choose a feature value The generative process for  follows  , [j]1Q, [c]1C}, where μjc and σjc are the mean and variance of cth component of jth Gaussian, respectively.The density function for X = , the optimal parameters α* and Ω* of MMNB-Gaussian model can be learned by maximizing the likelihood of the whole dataset  p, given byGiven a set of training sets l = logp for each testing event corresponding LNND descriptor X under the trained Fast MMNB. X is classified as an anomaly if the following criterion is satisfied:δ is a user defined threshold that controls sensitivity of the algorithm to anomaly detection. For dealing with the anomalies occurring in different scales, we perform anomaly detection at three scales of pyramid structures . In this work, we search 8 spatial nearest neighbors and 1 temporal nearest neighbor; that is, set K = 8 and N = 1, so the dimensionality of LNND descriptor is 9, and the topic number of Fast MMNB is set to 10.In this section, we validate the advantages of LNND descriptor by conducting extensive experiments on two public datasets including UCSD dataset and UMN dataset  to evaluate the performance for anomaly detection in UCSD dataset. The lower EER value is, the better performance is achieved. From The UMN dataset is captured from 3 different scenes, including indoor and outdoor scenes, and the resolution is 320 × 240. The abnormal event in the dataset is crowd panic escaping. They start with the normal event followed by the abnormal events. We portion each scene into two parts. The first part contains 400 frames and is used as training set which only contains normal events, and the rest is used as testing set which contains both normal and abnormal events. In the training stage, video sequences portioned the video into a set of cuboids with size of 10 × 10 × 5. In this paper, we propose a novel LNND descriptor to represent the video event for anomaly detection in crowded scenes. Compared with commonly used low-level feature descriptor in previous works, our LNND descriptor has two advantages. First, both the spatial and temporal contexts are incorporated into LNND descriptor. Using LNND descriptor, both the anomalous event and interactions between multiple events can be well detected by training a simple temporal model, unlike previous low-level feature descriptor which needs to rely on a spatiotemporal model to account for spatial context. Second, due to the low dimensionality of LNND descriptor, both the computation time and the memory requirement can be accordingly saved. We perform anomaly detection in UCSD and UMN datasets, and the results are provided for comparing with other state-of-the-art methods. The qualitative and quantitative analyses of experimental results demonstrate that our proposed LNND descriptor is a concise and efficient descriptor. Compared with commonly used low-level feature descriptor on previous anomaly detection works, our LNND-based method is computationally efficient and robust and has promising result. Meanwhile, the intermediate and subsequent process is less than most previous works. As our future work, we will attempt to use our proposed LNND descriptor to some other applications, such as event or action recognition.


Background. Despite the increased incidence of Kaposi sarcoma (KS) resulting from the Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS) pandemic, there is still significant underreporting of KS in this environment. Objectives. This study was aimed at determining the incidence and clinicopathologic patterns of KS among HIV infected patients in Lagos University Teaching Hospital (LUTH), Nigeria, over a 14-year period: January 2000 to December 2013. Methodology. The materials for this study included patients' hospital clinical files, duplicate copies of histopathologic reports, and tissue blocks and corresponding archival slides in the Anatomic and Molecular Pathology Department and the HIV/AIDS unit of the Department of Haematology. Results. Within the study period, 182 cases of KS were diagnosed, accounting for 1.2% of all patients managed for HIV/AIDS and 2.99% of solid malignant tumours. The male-to-female ratio and modal age group were 1 : 1.3 and 5th decade, respectively. Most cases (90%) had purely mucocutaneous involvement with the lower limb being the commonest site (65.8%). The majority of lesions were plaques (65.8%). Vascular formation was the predominant histologic type seen (43.5%). Conclusion. KS in Lagos followed the same epidemiologic trend as other centers in Nigeria, with an increasing incidence in this era of HIV/AIDS. Kaposi sarcoma (KS) is a vascular lesion of low-grade malignant potential that presents most frequently with skin lesions. It is the most common AIDS associated malignancy, and human herpes virus type 8 (HHV8) plays a significant role in its aetiopathogenesis , 2.A number of studies have reported both a change in the pattern and a dramatic increase in the incidence of KS as a consequence of the increased incidence of AIDS , 4. The Most reports from the northern part of Nigeria also indicate a change in the pattern of KS. In Jos, Mandong et al. reported 65 cases over 16 years (1987–2002) . More reThere is high prevalence of HIV/AIDS in Lagos as in other parts of Nigeria. While it might be assumed that the pattern of KS in this center conforms to the pattern seen in other centers and African countries, it is still necessary to get recent and accurate information about the disease in the country, if meaningful strategies for treatment and prevention are to be made.There has been evidence of an association of KS with second primary malignancies. Most often, patients with classic KS are at special risk of developing a malignancy of the lymphoreticular system . In a stWith improving care of people with HIV/AIDS and better management of related infections, especially tuberculosis, the longer survival of affected people in Nigeria will likely increase the burden of cancer as a clinical problem. However, reduction in the HIB/AIDS related KS may be an indicator of control of HIV/AIDS. The population's geographic and social diversity makes Nigeria a unique country for research on HIV/AIDS associated cancers. This can be embedded in programs targeting HIV/AIDS and other public health priorities. This study therefore was aimed at determining the incidence of KS among HIV infected patients in Lagos University Teaching Hospital. We also looked into the different clinical types of KS, their various histologic variants, and the impact of HIV/AIDS on the prevalence of KS.The materials for this study were derived from patients' hospital clinical files and request forms, duplicate copies of histopathological reports that were issued within the study period, and tissue blocks and corresponding archival slides in the Anatomic and Molecular Pathology Department of LUTH and the HIV/AIDS unit of the Department of Haematology, LUTH. All records of clinically diagnosed KS and those with histologically confirmed cases of KS from 2000 to 2013 were analyzed for age, sex, histologic variant, and the sites of distribution as well as their HIV/AIDS status. The frequency of the tumour was calculated relative to HIV/AIDS burden as well as all malignancies diagnosed in the same period. Histological classification was based on the morphological appearance of the tumour. Data were presented in figures, percentages, and simple frequency tables. The analysis of data from cases without histologic diagnoses served as a limitation of the study.The relative frequency is shown in The distribution of KS by site is shown in The most common lesions on the skin were plaques. These were seen in 108 of the patients and, of these, 67 (62%) had clinically equivocal lesions but were confirmed by histology while the diagnosis of the remaining 41 (38%) cases was based on clinical assessments. 29 cases had multiple nodular lesions involving only the lower limbs while 27 patients had a mixture of lesions including plaques, nodules, and ulcers involving multiple sites, especially the lower limbs.The histologic patterns are shown in Figures Three cases of childhood KS were seen among the histologically diagnosed cases: a 7-month-old boy with swellings on the head since birth, a 6-year-old girl with generalized nodular rashes, and a 10-year-old girl with generalized body rash.Only 182 cases of Kaposi sarcoma over a period of 14 years were recorded during the study period. This implies an average annual frequency of 13 cases. Lower incidence was earlier reported in the same center by Adeyi and Banjo who reported 8 cases of cutaneous KS over 6 years (1990 and 1995) . This fiKaposi sarcoma was not featured as one of the common cancers in Nigeria before the HIV/AIDS pandemic. In fact, most records at different cancer registries did not show cases of KS in their locality, probably because of its low occurrence. There was no case of KS in the records of cancer registry, UCH, Ibadan, before 1980 but between 1981 and 2000, 30 cases of KS were recorded in the center , 26. AltAlthough there is no previous similar study in the center of this study on the incidence of KS before the advent of AIDS pandemic, this study agreed with similar studies from other centers within the country and other African countries that, with the advent of AIDS pandemic, there has been an increase in the incidence of KS , 24. Pre+ T lymphocyte count.Seropositivity for KSHV in the general populace and among people infected with HIV is not known in this environment. This is an area for further study as the development of the tumour relates closely to the plasma titer value of the virus as well as the degree of immunosuppression as determined by the value of CD4Literature review revealed that the pattern of occurrence and trend of the disease differ from one country to another. In developed countries, for instance, AIDS associated KS has been seen most commonly among homosexual men, the incidence of which is declining. It is rarely seen among patients who have acquired HIV infection through heterosexual contact, intravenous drug use, or vertical transmission . In our Three cases of histologically diagnosed childhood KS with HIV infection were seen. This low incidence may be due to the possibility that childhood KS is not common in Nigeria or that a long latent time is required for the development of KS, which may be also influenced by medication. However, one has to be sure that cases of paediatric KS are not being missed or wrongly diagnosed as vascular malformations, for example, haemangioma, granuloma pyogenicum, granulation tissue, popular urticaria, and inflammatory dermatoses. In Uganda, the incidence of childhood Kaposi sarcoma has risen more than 40-fold in the era of HIV/AIDS . A high KS is not a clinical diagnosis as various lesions such as melanoma, mycoses, and Madura foot, which are also common on the leg, could mimic KS. Since these entities have also been reported in patients with HIV, obtaining a histologic confirmation of KS is important. However, this poses a significant financial constraint to many patients undergoing treatment for HIV in our environment. This lack of histologic confirmation in a proportion of cases served as a limitation for our study.Histopathologists are familiar with the histological features of usual  KS as it progresses from patch to plaque and finally nodular stages –33. ThisIn this study, the histological patterns of KS observed, as shown in Predominantly vascular formation shown in Inflammatory response is commonly associated with ulcerated skin lesions and this was seen in 75% of cases irrespective of the pattern. Inflammatory response giving the appearance of granulation tissue can lead to misdiagnosis of KS especially in early lesions. In cases of KS, the endothelial cell lining is large and protrudes into the lumen and the inflammatory cell infiltrates are of varying intensity and are perivascular or diffusely disposed. Also, the presence of extravasated red blood cells and hemosiderin always indicates the possibility of early KS.Failure to identify a given lesion as KS could lead to wrong or delayed diagnosis and hence inappropriate management. Immunohistochemistry should be utilized in controversial cases.Majority of the cases of Kaposi sarcoma in this study were mucocutaneous and many of the cases were multifocal involving multiple sites . In most of these cases, the lower limbs were mostly involved. This observation is in agreement with other reports on KS. One would expect more cases of KS in this setting to involve the viscera because of late presentation and association with HIV. In resource poor centers where sophisticated diagnostic techniques such as CT scan and fibre-optic endoscopic examination are not available, clinical examination supported with plain chest X-ray could help to assess visceral involvement.KS in Lagos follows the same epidemiologic trend as other centers in Nigeria, with an increasing incidence in this era of HIV/AIDS. Most cases are mucocutaneous and occur at multiple sites, with the lower limbs being the commonest site of involvement.


Dendritic cells (DC) play a critical role in the initiation of the immune response, acting as sentinels in the tissues, reacting to invading pathogens and then inducing the activation and differentiation of naïve T cells. Since their discovery by Ralph Steinman in the 1970s, we have learned a great deal about DC biology and function. Distinct DC subsets exist in specific tissue niches and within the secondary lymph nodes, and the study of the phenotype and function of DC subsets in mice and humans has been an area of great interest. DC influence the immune response by directing the differentiation of T cells into functional subtypes important for elimination of the relevant pathogen. DC also contribute to disease pathogenesis, such as autoimmunity and cancer, through failures in self-tolerance and promotion of an immunosuppressive environment, respectively. In addition, because of their role in molding T cell responses, DC have been tested in therapeutic settings in both autoimmunity and cancer. In this research topic, 11 articles cover many aspects of current DC biology, ranging from the classification and function of DC subsets, roles of DC in disease pathogenesis to current use of DC in the therapeutic setting.The topic begins with two reviews of normal DC function; one is focused on the important topic of cross-presentation  and the in vivo and in vitro (The next four articles in this research topic are original research focusing on various aspects of DC function in vitro –6. DC main vitro  examinedin vitro . DC playin vitro  demonstrin vitro . This paThe final group of five reviews highlights the role of DC in either causing or ameliorating disease –11. AutoThus, this research topic provides a timely overview of some of the recent advances in DC biology and we look forward to many new developments in this exciting field.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Dopamine neurons in the midbrain have a central role in generating cycles of biological activity with periods as short as 4 hours and as long as 100 hours. Related research article Blum ID, Zhu L, Moquin L, Kokoeva MV, Gratton A, Giros B, Storch KF. 2014. A highly-tunable dopaminergic oscillator generates ultradian rhythms of behavioral arousal. eLife3:e05105. doi: 10.7554/eLife.05105Image Methamphetamine increases the period of ultradian activity rhythms in Bmal1 null mice by increasing dopamine levelsThe rotation of the earth as it orbits around the sun creates daily cycles of light, temperature and other factors that are vitally important for myriad biological processes. Indeed, almost all organisms exhibit circadian gene expression—that is, certain genes are expressed as proteins at a rate that oscillates with a period of about 24 hr—and associated biochemical and behavioral rhythms. Circadian rhythms are the organization of biological activities into cycles that oscillate with a period of around 24 hours, and we now know a great deal about the molecular and neural bases of these rhythms . MoreoveeLife, Ian Blum and Kai-Florian Storch of McGill University and the Douglas Institute, both in Montreal, and co-workers report that dopamine—a chemical in the brain that is usually associated with pleasure and motivated reward—has a central role in setting the frequency of ultradian rhythms and may also account for rhythmic phenomena with longer periods in the circadian and infradian ranges  or greater than  24 hours. The underlying mechanisms for most of these rhythms are poorly understood. Now, in n ranges .Bmal1 had been deleted, still maintained an activity rhythm with a period of approximately 4 hours. The link to dopamine was made when they tested mice lacking both the SCN and the dopamine reuptake transporter . In these mice, ultradian rhythms lengthened from 4 hours to 12 hours. A similar effect was obtained by administering methamphetamine, a stimulant that also increases synaptic dopamine: indeed, the period increased with the dose, reaching 100 hours at the highest doses.In mammals, circadian rhythms can be eliminated by ablation of the suprachiasmatic nucleus (SCN) in the brain or by the deletion of so-called circadian clock genes. The starting point for Blum et al. was the observation that mice lacking a circadian rhythm because their SCN had been ablated, or because the clock gene Bmal1 gene had been deleted. They found, as expected, that increased activation of dopamine neurons increased the period of the ultradian rhythms. These data reveal for the first time a neurochemical that regulates the rate at which ultradian rhythms cycle. From these observations, Blum et al. speculate that dysregulation of dopamine signaling in the human brain could amplify and lengthen ultradian rhythms, resulting in cycles of mood and motivation with periods of 24 hours or longer, such as those characteristic of depression and bipolar disorder.To directly test the hypothesis that ultradian rhythms are driven by a ‘dopamine ultradian oscillator’, Blum et al. measured the level of dopamine in several contexts. They found that the dopamine level in a region of the brain called the dorsal striatum rose and fell in synchrony with the ultradian rhythm. Moreover, they used a chemogenetic approach to directly manipulate the dopamine neurons in the midbrain of mice in which the SCN had been ablated and the The dopamine ultradian oscillator concept may also be relevant to two other rhythms that operate independently of the circadian pacemaker in the SCN. One of these is the effect of methamphetamine: as mentioned above, high doses of this stimulant induce infradian rhythms that uncouple from the circadian rhythm in wild-type rodents. These rhythms, previously attributed to a ‘methamphetamine sensitive circadian oscillator’ , can nowBmal1. The search for the neural substrates of these food anticipatory rhythms has focused on the hypothalamus, but there is growing evidence that dopamine neurons in the midbrain may be important. This was first suggested by observations that circadian anticipatory rhythms can be induced by restricted access to sex, water or palatable treats. The dopamine neurons in the midbrain project to the ventral and dorsal striatum, and both of these areas exhibit circadian rhythms in the expression of clock genes . However, the circadian constraint is lost in ntervals . These fntervals .Some of these oscillators are not constrained by circadian clock genes, and they regulate ultradian cycles of foraging (and other activities) that can lengthen to infradian cycles under high levels of dopamine. Other oscillators are constrained by circadian clock genes and control circadian cycles of foraging activity when food availability recurs predictably at 24-hour intervals. All of these oscillators may be dopamine sensitive, and conceivably may reside in midbrain dopamine neurons, the dorsal striatum, or both.The same population of oscillators may also be involved in other manifestations of timing behavior, such as the ability to sense the passage of time in the seconds to minutes range . Impaired time perception is another manifestation of psychiatric disorders, which leads back to the suggestion by Blum et al. that dopamine ultradian oscillators in the striatum may be critical for understanding mood cycling and thought disorders.What remains to be established is whether the timing mechanisms responsible for ultradian rhythms are located entirely within midbrain dopamine neurons and the dorsal striatum, or are distributed more broadly, and how these mechanisms operate at the molecular level. From these answers may emerge new treatments for disorders of temporal organization that may underpin mental illness.


This case highlights that measurement of HHV8 load in plasma may be useful to identify patients at risk for KS-IRIS, and that this parameter should be included in the design of larger studies to define KS-IRIS risk predictors.We present here a case of immune reconstitution inflammatory syndrome associated with Kaposi’s sarcoma (KS-IRIS) developed in an AIDS patient two months after initiation of antiretroviral therapy (ART). Baseline characteristics of this IRIS-KS case, within a cohort of 12 naïve AIDS-KS patients, were analyzed. No statistically significant differences in CD4 cell counts, plasma HIV RNA load, KS clinical staging, human herpesvirus 8 (HHV8) antibody titers and HHV8 load in peripheral blood mononuclear cells and saliva were evidenced. HHV8 load in plasma was found to be significantly higher in the KS-IRIS patient (> 6 log Within a further 4 week-treatment, the patient showed recrudescent KS, with an increased number of lesions (> 20 cutaneous lesions) and enlargement of pre-existing ones. CD4 cell count was 820 cells/ml (21 %) and HIV RNA viral load was under the limit of detectability. The sudden clinical KS worsening together with the rapid immunovirological response was indicative of an immune reconstitution inflammatory syndrome rather than KS progression. The patient continued the antiretroviral combination therapy and no systemic KS chemotherapy was administrated. KS-IRIS flare completely recovered in 10 months.A 39-year-old HIV-positive homosexual male was admitted to our Infectious Diseases Unit with asthenia and multiple 11) painless, nodular purple lesions concentrated in the trunk that ranged from 0.5 to 2.0 cm in the largest dimension. The patient was diagnosed with HIV infection and cutaneous Kaposi’s sarcoma was confirmed by histological examination. Either mucocutaneous and visceral involvement was excluded by endoscopic and neck, chest and abdominal computerized tomography (CT) examinations. The CD4 lymphocyte count was 360 cells/μl (14 %) and plasma HIV-1 RNA level was 4.09 log1 painlesth–75th percentile) calculated on the measurable samples. To compare the values of different biomarkers of the KS-IRIS case to those of the KS group, the one-tailed Crawford and Howell t-test was used [10 GE/ml, p = 0.01) compared to those measured in patients with KS regression. Moreover, HHV8 load in saliva was higher, albeit not statistically significant (p = 0.09), in the KS-IRIS case  of plasma HHV8 load. We think that these discrepancies might be due to different sensitivities in the quantitative techniques, if samples were independently reanalyzed, or to a unrecognized bias related to the selection of the 104 patients for the pooled analysis. Another population of AIDS-KS patients included in Letang et al. [10 copies/ml [per se cannot be a predictive parameter.These findings, although limited to a small cohort, indicate that plasma HHV8 DNA detectability may not be so informative, and are fully in line with previously published studies. Indeed, 78.4 % of ART-naïve AIDS-KS patients from Zimbabwe had measurable plasma load, with a baseline median value of 2.82 logopies/ml , 7. Moreopies/ml . The samopies/ml , and plag et al.  is the Dg et al. ; about hopies/ml . TherefoThis case highlights that, in our small cohort, the measurement of baseline HHV8 load in plasma was useful to identify the patient who experienced a KS-IRIS. Therefore, we recommend including quantification of baseline parameters linked to HHV8 infection status, specifically measurement of HHV8 load in plasma and, possibly, in saliva, in the design of larger studies to define KS-IRIS predictors.This study was evaluated and approved by the Ethical Committee of the Veneto Institute of Oncology (Protocol n. 14256), and written informed consent was obtained from study participants.