Gemcitabine, carboplatin, and Epstein-Barr virus-specific autologous cytotoxic T lymphocytes for recurrent or metastatic nasopharyngeal carcinoma: VANCE, an international randomized phase III trial.

BACKGROUND: Epstein-Barr virus-specific cytotoxic T lymphocyte (EBV-CTL) is an autologous adoptive T-cell immunotherapy generated from the blood of individuals and manufactured without genetic modification. In a previous phase II trial of locally recurrent or metastatic nasopharyngeal carcinoma (R/M NPC) patients, first-line gemcitabine and carboplatin (GC) and EBV-CTL combination demonstrated objective antitumor EBV-CTL activity and a favorable safety profile. The present study explored whether this combined first-line chemo-immunotherapy strategy would produce superior clinical efficacy and better quality of life compared with conventional chemotherapy treatment. PATIENTS AND METHODS: This multicenter, randomized, phase III trial evaluated the efficacy and safety of GC followed by EBV-CTL versus GC alone as first-line treatment of R/M NPC patients. Thirty clinical sites in Singapore, Malaysia, Taiwan, Thailand, and the USA were included. Subjects were randomized to first-line GC (four cycles) and EBV-CTL (six cycles) or GC (six cycles) in a 1 : 1 ratio. The primary outcome was overall survival (OS) and secondary outcomes included progression-free survival, objective response rate, clinical benefit rate, quality of life, and safety. CLINICALTRIALS: gov identifier: NCT02578641. RESULTS: A total of 330 subjects with NPC were enrolled. Most subjects in both treatment arms received four or more cycles of chemotherapy and most subjects in the GC + EBV-CTL group received two or more infusions of EBV-CTL. The central Good Manufacturing Practices (GMP) facility produced sufficient EBV-CTL for 94% of GC + EBV-CTL subjects. The median OS was 25.0 months in the GC + EBV-CTL group and 24.9 months in the GC group (hazard ratio = 1.19; 95% confidence interval 0.91-1.56; P = 0.194). Only one subject experienced a grade 2 serious adverse event related to EBV-CTL. CONCLUSIONS: GC + EBV-CTL in subjects with R/M NPC demonstrated a favorable safety profile but no overall improvement in OS versus chemotherapy. This is the largest adoptive T-cell therapy trial reported in solid tumors to date.

Diversity and distribution of ectoparasite taxa associated with Micaelamys namaquensis (Rodentia: Muridae), an opportunistic commensal rodent species in South Africa.

South Africa boasts a rich diversity of small mammals of which several are commensal and harbour parasites of zoonotic importance. However, limited information is available on the parasite diversity and distribution associated with rodents in South Africa. This is particularly relevant for Micaelamys namaquensis (Namaqua rock mouse), a regionally widespread and locally abundant species that is often commensal. To address the paucity of data, the aims of the study were to record the ectoparasite diversity associated with M. namaquensis and develop distribution maps of lice and mites associated with M. namaquensis and other rodents in South Africa. Micaelamys namaquensis individuals (n = 216) were obtained from 12 localities representing multiple biomes during 2017­2018. A total of 5591 ectoparasites representing 5 taxonomic groups ­ fleas, lice, mesostigmatid mites, chiggers and ticks was recorded. These consisted of at least 57 taxa of which ticks were the most speciose (20 taxa). Novel contributions include new host and locality data for several ectoparasite taxa and undescribed chigger species. Known vector species were recorded which included fleas (Ctenocephalides felis, Dinopsyllus ellobius and Xenopsylla brasiliensis) and ticks (Haemaphysalis elliptica, Rhipicephalus appendiculatus and Rhipicephalus simus). Locality records indicate within-taxon geographic differences between the 2 louse species and the 2 most abundant mite species. It is clear that M. namaquensis hosts a rich diversity of ectoparasite taxa and, as such, is an important rodent species to monitor in habitats where it occurs in close proximity to humans and domestic animals.

Microbial metabolomics: essential definitions and the importance of cultivation conditions for utilizing Bacillus species as bionematicides.

Root-knot nematodes are destructive phytopathogens that damage agricultural crops globally, and there is growing interest in the use of biocontrol based on rhizobacteria such as Bacillus to combat Meloidogyne species. It is hypothesized that nematicidal activity of Bacillus can be attributed to the production of secondary metabolites and hydrolytic enzymes. Yet, few studies have characterized these metabolites and their identities remain unknown. Others are speculative or fail to elaborate on how secondary metabolites were detected or distinguished from primary metabolites. Metabolites can be classified based on their origin as either intracellular or extracellular and based on their function, as either primary or secondary. Although this classification is in general use, the boundaries are not always well defined. An understanding of the secondary metabolite and hydrolytic enzyme classification of Bacillus species will facilitate investigations aimed at bionematicide development. This review summarizes the significance of Bacillus hydrolytic enzymes and secondary metabolites in bionematicide research and provides an overview of known classifications. The importance of appropriate cultivation conditions for optimum metabolite and enzyme production is also discussed. Finally, the use of metabolomics for the detection and identification of nematicidal compounds is considered.

Helminth parasites of impalas, Aepyceros melampus, in eastern southern Africa, collected during 1973 to 2007.

This paper summarises published and unpublished data on helminths collected systematically from 424 impalas at 11 localities in eastern southern Africa, from St. Lucia in KwaZulu-Natal (KZN) to the Tuli Block in north-eastern Botswana. It includes data on collections in the Kruger National Park (KNP) in the drought of 1982, and in 1992/93 following the 1991/92 drought. Thirty-three species of nematodes, plus six taxa identified only to the generic level, three taxa of trematodes, and three species of cestodes were collected. Helminth species richness was highest in the southern KNP and lowest in the Tuli Block. The prevalence and intensity of infection of several helminths also declined from KZN and the southern KNP to the drier areas in the north and west. With the exception of St. Lucia and Nylsvley, > 80% of the helminths collected at each locality were collected in the southern KNP. St. Lucia was the most dissimilar locality; of the 20 helminths collected, five were unique. Ten of 33 species of nematodes, the paramphistomines (Trematoda) and the cestode Stilesia hepatica were collected at seven or more localities. Six of the most common nematodes, Cooperia hungi, Cooperioides hamiltoni, Impalaia tuberculata, Longistrongylus sabie, Strongyloides papillosus and Trichostrongylus deflexus are primarily parasites of impalas in the southern KNP, whereas many of the helminths collected at only one or two localities are parasites of other hosts. Nematode burdens were increased in the drought affected impalas in 1982, but helminth burdens decreased in 1992/93 following a dry cycle.

Displacement of Rhipicephalus decoloratus by Rhipicephalus microplus (Acari: Ixodidae) in the Eastern Cape Province, South Africa.

The objective of the study was to establish to what extent the native tick species Rhipicephalus decoloratus has been displaced by the invasive introduced tick, Rhipicephalus microplus at two communally grazed areas in the Eastern Cape Province, South Africa. To this end ticks were collected monthly from five cattle over a period of 2 years and from 10 drag-samples of the vegetation over a period of 1 year at each locality. Whereas 10 years previously only R. decoloratus and no R. microplus had been recorded in the vicinity of the two sites, R. microplus now comprised the bulk of collections at both. Furthermore, significantly more R. microplus were collected from cattle at both localities during the 2nd year of the survey than during the 1st. In addition to 83 instances of intraspecific coupling, there were 17 instances of R. microplus males coupled with R. decoloratus females. Collections made from cattle and goats on two farms close to the study sites revealed that R. microplus was present on both host species and that it significantly outnumbered R. decoloratus on one of the farms. Rhipicephalus decoloratus and R. microplus larvae as well as larvae exhibiting characteristics of both species were collected from the vegetation.

Abiotic and biotic determinants of tick burdens in the eastern rock sengi (Elephantulus myurus).

Ticks are important vectors of pathogens of medical and veterinary importance worldwide. In spite of their economic importance, our current knowledge about the factors affecting tick prevalence and abundance in tropical and subtropical regions is rather limited. Both abiotic (e.g. temperature) as well as biotic variables (e.g. host sex) have been identified as key determinants of distributions. Eastern rock sengis or elephant shrews (Elephantulus myurus, Macroscelidea: Cacroscelididae, Thomas & Schwann) are widely distributed throughout Africa and can harbour a large number of tick species and substantial tick burdens. In the present study, we evaluated the contribution of climate and host factors on tick burdens of sengis. Throughout the year sengis carried high abundances of immature stages of a single tick species, Rhipicephalus sp. near warburtoni. There was no evidence that host parameters affected tick burdens. However, larval abundance decreased with increasing ambient temperatures and both larvae and nymphs were negatively affected by rainfall 2 months before the sampling month. In addition, nymphal burdens decreased with increasing minimum temperatures. Our results suggest that climate factors are the largest constraint for the immature stages of R. sp. near warburtoni and that eastern rock sengis could play a crucial role in the dynamics of tick-borne diseases as a result of the large tick burdens they can sustain.

ADAPTOR PROTEIN Ruk/CIN85REGULATES REDOX BALANCE IN 4T1MOUSE BREAST CANCER CELLS EXPOSED TO PLASMIN(OGEN).

BACKGROUND: Cell surface plasmin is involved in tumor growth and metastatic dissemination by regulating cancer cells adhesion, migration and invasion. Plasmin-induced cell detachment is accompanied by an increased rate of reactive oxygen species (ROS) generation and cell death. However, cancer cells acquire the ability to develop adaptive mechanisms to resist ROS-mediated apoptosis. AIM: To establish the role of adaptor protein Ruk/CIN85 in the control of viability and redox balance in breast adenocarcinoma cells exposed to plasmin(ogen). MATERIALS AND METHODS: Mouse 4T1 cells with the stable overexpression of adaptor protein Ruk/CIN85 (RukUp subline) and corresponding control (Mock subline) were treated with Glu-plasminogen (1-100 nM). Plasminogen to plasmin conversion was monitored spectrophotometrically by cleavage of the specific chromogenic substrate S2251. Specific uPA inhibitor BC11 was used to verify the uPA-mediated mechanism of plasminogen pericellular activation by 4T1 cells. Cell survival rate was assessed by MTT-test and cell proliferation was estimated by colony formation assay. Enzymatic activities of catalase, glutathione peroxidase, superoxide dismutase, as well as hydrogen peroxide (H2O2) levels were measured by spectrophotomertric and fluorometric assays. The intracellular ROS generation was monitored by flow cytometry using H2DCF-DA fluorescent probe. RESULTS: Plasminogen was shown to be converted into an active proteinase plasmin on the surface of carcinoma cells in uPA-dependent manner. Plasmin(ogen) suppressed proliferation and affected survival of both studied 4T1 sublines. However, RukUp cells displayed higher resistance to plasmin(ogen)-induced cytotoxicity than Mock cells. Plasmin(ogen) promoted significant elevation in ROS generation rate in cells with the basal level of Ruk/CIN85 expression. In contrast, RukUp cells appear to be more effective in counteracting prooxidant changes due to the activation of some enzymes of the glutathione system, in particular glutathione peroxidase, and a concomitant decrease of H2O2 accumulation. CONCLUSION: Adaptor protein Ruk/CIN85 is involved in the regulation of redox homeostasis in cancer cells to maintain levels of ROS, thus promoting redox adaptation in cancer cells exposed to plasmin(ogen). Thus, Ruk/CIN85 may represent one of the relevant targets in order to diminish the resistance of cancer cells to ROS-mediated apoptosis.

Down-modulation of cancer targets using locked nucleic acid (LNA)-based antisense oligonucleotides without transfection.

Usually, small interfering RNAs and most antisense molecules need mechanical or chemical delivery methods to down-modulate the targeted mRNA. However, these delivery approaches complicate the interpretations of biological consequences. We show that locked nucleic acid (LNA)-based antisense oligonucleotides (LNA-ONs) readily down-modulate genes of interest in multiple cell lines without any delivery means. The down-modulation of genes was quick, robust, long-lasting and specific followed by potent down-modulation of protein. The efficiency of the effect varied among the 30 tumor cell lines investigated. The most robust effects were found in those cells where nuclear localization of the LNA-ON was clearly observed. Importantly, without using any delivery agent, we demonstrated that HER3 mRNA and protein could be efficiently down-modulated in cells and a tumor xenograft model. These data provide a simple and efficient approach to identify potential drug targets and animal models. Further elucidation of the mechanism of cellular uptake and trafficking of LNA-ONs may enhance not only the therapeutic values of this platform but also antisense molecules in general.

T cells, but not B cells, are required for bowel inflammation in interleukin 2-deficient mice.

Interleukin-2 (IL-2)-deficient (IL-2-/-) mice develop anemia and colonic inflammatory bowel disease. To elucidate the mechanism of this disease, we have bred IL-2-/- mice to two strains of immunodeficient mice, RAG-2-deficient (RAG-2-/-, lacking B and T cells) and JH-deficient mice (JH-/-, lacking B cells). IL-2-/-, RAG-2-/- double-mutant mice are disease free, while IL-2-/-, JH-/- double-mutant mice succumb to bowel disease at the same rate as IL-2-/- littermates. IL-2-/-, JH-/- mice do not, however, succumb to anemia. Thus, spontaneous intestinal inflammation in IL-2-/- mice requires mature T cells, not B cells, while anemia is dependent on B cells.

Crucial role of interferon consensus sequence binding protein, but neither of interferon regulatory factor 1 nor of nitric oxide synthesis for protection against murine listeriosis.

Listeria monocytogenes is widely used as a model to study immune responses against intracellular bacteria. It has been shown that neutrophils and macrophages play an important role to restrict bacterial replication in the early phase of primary infection in mice, and that the cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) are essential for protection. However, the involved signaling pathways and effector mechanisms are still poorly understood. This study investigated mouse strains deficient for the IFN-dependent transcription factors interferon consensus sequence binding protein (ICSBP), interferon regulatory factor (IRF) 1 or 2 for their capacity to eliminate Listeria in vivo and in vitro and for production of inducible reactive nitrogen intermediates (RNI) or reactive oxygen intermediates (ROI) in macrophages. ICSBP-/- and to a lesser degree also IRF2-/- mice were highly susceptible to Listeria infection. This correlated with impaired elimination of Listeria from infected peritoneal macrophage (PEM) cultures stimulated with IFN-gamma in vitro; in addition these cultures showed reduced and delayed oxidative burst upon IFN-gamma stimulation, whereas nitric oxide production was normal. In contrast, mice deficient for IRF1 were not able to produce nitric oxide, but they efficiently controlled Listeria in vivo and in vitro. These results indicate that (a) the ICSBP/IRF2 complex is essential for IFN-gamma-mediated protection against Listeria and that (b) ROI together with additional still unknown effector mechanisms may be responsible for the anti-Listeria activity of macrophages, whereas IRF1-induced RNI are not limiting.

Ixodid ticks on domestic dogs in the Northern Cape Province of South Africa and in Namibia.

The objective of this study was to determine the species composition of ixodid ticks infesting domestic dogs in the northwestern region of the Northern Cape Province of South Africa and in Namibia. Ticks were collected from February 2008 to January 2009 from dogs presented for a variety of reasons at a veterinary clinic in the Northern Cape Province and at 3 clinics in Namibia. The ticks collected at each place were pooled separately for each month at each locality. Eleven ixodid tick species were collected from dogs in the Northern Cape Province and new locality records for Haemaphysalis colesbergensis and Ixodes rubicundus, new locality and host records for Hyalomma glabrum, and a new host record for Rhipicephalus neumanni are reported. Six tick species were collected from dogs at the 3 clinics in Namibia. The most numerous species on dogs in both countries was R. sanguineus. The present results increase the total number of ixodid tick species collected from dogs in South Africa from 25 to 28.

The efficacy of a topically applied combination of cyphenothrin and pyriproxyfen against the southern African yellow dog tick, Haemaphysalis elliptica, and the cat flea, Ctenocephalides felis, on dogs.

The objective of this study was to determine the therapeutic and residual efficacy of a topically applied combination of cyphenothrin (40%) and pyriproxyfen (2%) against the tick Haemaphysalis elliptica and the flea Ctenocephalides felis on dogs. Twelve dogs were infested with 50 ticks 2 days before they were treated and with approximately 100 fleas 6 days before treatment and again 2 days before treatment and with 50 ticks and approximately 100 fleas at weekly intervals thereafter. They were ranked according to their flea counts and sex 5 days before treatment and randomly allocated to an untreated control group of 6 dogs and a treated group of 6 dogs. Ticks and fleas were collected from the dogs 48 h after treatment and 48 h after each infestation and live and dead ticks and live fleas were counted. The counts of ticks and fleas were transformed to geometric means, and efficacy was calculated by comparing these means. The product had a therapeutic efficacy of 83.1% against H. elliptica and 97.5% against C. felis 2 days after treatment. The residual period of protection during which efficacy was > or = 90% was 5 weeks for both H. elliptica and C. felis.

Progressive encephalopathy and myopathy in transgenic mice expressing human foamy virus genes.

Transgenic mice carrying the bel region of human foamy retrovirus (HFV) under transcriptional control of its own long terminal repeat expressed the transgene in their central nervous systems and in smooth and striated muscle tissues. The animals developed a progressive degenerative disease of the central nervous system and of the striated muscle. Because expression of the transgene was closely correlated with the appearance of structural damage and inflammatory reactions were scanty, the disease is likely to be caused directly by the HFV proteins. These unexpected findings call for a reevaluation of the pathogenic potential of HFV in humans.

Immune responses in interleukin-2-deficient mice.

The role of costimulatory signals in T cell induction was evaluated in mice lacking the interleukin-2 (IL-2) gene. In vitro secondary antiviral T cell responses were absent unless IL-2 was added, confirming the crucial role of IL-2 in vitro. In vivo, primary and secondary cytotoxic T cell responses against vaccinia and lymphocytic choriomeningitis virus were within normal ranges. B cell reactivity to vesicular stomatitis virus was not impaired. T helper cell responses were delayed but biologically functional. Natural killer cell activity was markedly reduced but inducible. These normal in vivo immune responses in IL-2-deficient mice question the importance of IL-2 as defined by in vitro studies.

A century of tick taxonomy in South Africa.

Eighty ixodid tick species, 25 argasid tick species and Nuttalliella namaqua occur in South Africa. Twenty-one of the 80 ixodid species and two of the argasid species occur only in this country, while N. namaqua is present only in South Africa and Namibia. Forty-six of the 80 ixodid species and 16 of the 25 argasid species as well as N. namaqua have been described as new species since 1908. People working in South Africa have written or contributed to the descriptions of 24 of these 63 new species, while foreign researchers have described the remainder. New species indigenous to South Africa are still being discovered, while the names of some species, well known because of their veterinary importance, have been altered.

Species composition and geographic distribution of ticks infesting cattle, goats and dogs in a temperate and in a subtropical region of south-east Africa.

The species and distribution of ticks infesting cattle, goats and dogs in the eastern region of the Eastern Cape Province, South Africa and Maputo Province, Mozambique were determined from collections made from these animals at 72 localities in the former region and 30 in the latter. Eleven ixodid and one argasid species were recovered in the Eastern Cape Province and 15 ixodid species in Maputo Province. The most common ticks infesting cattle and goats in both provinces were Amblyomma hebraeum, Rhipicephalus (Boophilus) microplus, Rhipicephalus appendiculatus and Rhipicephalus evertsi evertsi. The dominant species on dogs were Haemaphysalis elliptica and Rhipicephalus simus. The geographic distributions of the major species and some of the minor species in both regions were plotted. The partial or complete displacement of the indigenous tick Rhipicephalus (Boophilus) decoloratus by the introduced species R. (B.) microplus was a major feature of both surveys.

Ticks collected from birds in the northern provinces of South Africa, 2004-2006.

Approximately 3000 birds, mainly passerines, caught in mist nets in the northern provinces of South Africa, were examined for ticks. A total of 178 ticks, belonging to 14 species, were recovered from 83 birds of 43 different species. Hyalomma rufipes was the most numerous tick, with 26 larvae and 109 nymphs collected, followed by Amblyomma marmoreum, with 13 larvae and two nymphs. Despite the study being conducted within the distribution range of Amblyomma hebraeum, it was not seen on any passerines, whereas three larger species were infested. The potential for small birds to spread ticks with their associated tick-borne pathogens is discussed.

The comparative prevalence of five ixodid tick species infesting cattle and goats in Maputo Province, Mozambique.

This study compares the prevalence of ixodid tick species on cattle and goats in Maputo Province. Adult ticks as well as the nymphs of three species, and only the adults of two species were collected from sets of five cattle at 21 localities throughout the province and compared with those collected from similar sets of goats at the same places. Amblyomma hebraeum adults and/or nymphs were present on cattle and on goats at all 21 localities, and 90 cattle and 22 goats were infested with adult ticks. Rhipicephalus (Boophilus) microplus adults and/or nymphs were collected from cattle at 20 and from goats at 15 localities, and 92 cattle and 34 goats were infested [Chi-square test (chi2), P<0.001]. The total length of several maturing female R. (Boophilus) microplus collected from cattle and goats exceeded 5 mm, indicating that they successfully engorge on both host species. Rhipicephalus appendiculatus adults and/or nymphs were present on cattle at 15 and on goats at 13 localities, but 28 cattle and only one goat were infested with adult ticks (chi2, P<0.001). Adult Rhipicephalus evertsi evertsi were recovered from cattle at 20 and from goats at 17 localities, and 74 cattle and 69 goats were infested. Adult Rhipicephalus simus were collected from cattle at 18 and from goats at 11 localities (Fisher's exact test, P=0.04), and 60 cattle and 14 goats were infested (chi2, P<0.001). These findings underscore the advisability of including goats in acaricide application programmes designed for the control of tick-borne diseases in cattle at the same locality.

The prevalence and distribution of Argas walkerae (Acari: Argasidae) in the eastern region of the Eastern Cape Province, South Africa.

The prevalence and geographic distribution of the fowl tampan, Argas walkerae Kaiser & Hoogstraal, 1969 was determined in the eastern region of the Eastern Cape Province, South Africa by inspecting two fowl houses in the vicinity of each of 72 randomly selected communal cattle dip-tanks. Tampans were collected from 102 (70.8%) of the 144 fowl houses in the neighbourhood of 57 (79.2%) of the 72 selected dip-tanks, and the localities of the collections were mapped. Argas walkerae was present in fowl houses from the warm coastal regions of the Indian Ocean in the south to the cold and mountainous Drakensberg in the north-east of the Province. Taking into account the probable sensitivity of the sampling method, it is estimated that A. walkerae is likely to be present in fowl houses belonging to between 74 and 84% of communities making use of cattle dip-tanks in the eastern region of the Eastern Cape Province, and that when it is present, between 64 and 75% of fowl houses will be infested. The geographic distribution of A. walkerae seemed to be more strongly associated with the presence of fowls and fowl houses containing raw or processed wood in their structure than with climate.

Ixodid ticks on dogs belonging to people in rural communities and villages in Maputo Province, Mozambique.

The species composition and geographic distribution of ixodid ticks infesting domestic dogs owned by people in rural communities and villages in Maputo Province was established by collecting ticks from dogs at each of 27 localities spread throughout the province. Ticks were collected from a total of 132 dogs, and nine species belonging to four genera were identified. One dog was infested with six species, three with five and 13 with four species. Haemaphysalis elliptica followed by Rhipicephalus simus were present on dogs at most localities, and their geographic distribution in Maputo Province has been mapped for the first time.