Dynamics of digestive proteolytic system during blood feeding of the hard tick Ixodes ricinus

<p>Abstract</p> <p>Background</p> <p>Ticks are vectors of a wide variety of pathogens causing severe diseases in humans and domestic animals. Intestinal digestion of the host blood is an essential process of tick physiology and also a limiting factor for pathogen transmission since the tick gut represents the primary site for pathogen infection and proliferation. Using the model tick <it>Ixodes ricinus</it>, the European Lyme disease vector, we have previously demonstrated by genetic and biochemical analyses that host blood is degraded in the tick gut by a network of acidic peptidases of the aspartic and cysteine classes.</p> <p>Results</p> <p>This study reveals the digestive machinery of the <it>I. ricinus </it>during the course of blood-feeding on the host. The dynamic profiling of concentrations, activities and mRNA expressions of the major digestive enzymes demonstrates that the <it>de novo </it>synthesis of peptidases triggers the dramatic increase of the hemoglobinolytic activity along the feeding period. Overall hemoglobinolysis, as well as the activity of digestive peptidases are negligible at the early stage of feeding, but increase dramatically towards the end of the slow feeding period, reaching maxima in fully fed ticks. This finding contradicts the established opinion that blood digestion is reduced at the end of engorgement. Furthermore, we show that the digestive proteolysis is localized intracellularly throughout the whole duration of feeding.</p> <p>Conclusions</p> <p>Results suggest that the egressing proteolytic system in the early stage of feeding and digestion is a potential target for efficient impairment, most likely by blocking its components via antibodies present in the host blood. Therefore, digestive enzymes are promising candidates for development of novel 'anti-tick' vaccines capable of tick control and even transmission of tick-borne pathogens.</p

Profiling of proteolytic enzymes in the gut of the tick Ixodes ricinus reveals an evolutionarily conserved network of aspartic and cysteine peptidases

<p>Abstract</p> <p>Background</p> <p>Ticks are vectors for a variety of viral, bacterial and parasitic diseases in human and domestic animals. To survive and reproduce ticks feed on host blood, yet our understanding of the intestinal proteolytic machinery used to derive absorbable nutrients from the blood meal is poor. Intestinal digestive processes are limiting factors for pathogen transmission since the tick gut presents the primary site of infection. Moreover, digestive enzymes may find practical application as anti-tick vaccine targets.</p> <p>Results</p> <p>Using the hard tick, <it>Ixodes ricinus</it>, we performed a functional activity scan of the peptidase complement in gut tissue extracts that demonstrated the presence of five types of peptidases of the cysteine and aspartic classes. We followed up with genetic screens of gut-derived cDNA to identify and clone genes encoding the cysteine peptidases cathepsins B, L and C, an asparaginyl endopeptidase (legumain), and the aspartic peptidase, cathepsin D. By RT-PCR, expression of asparaginyl endopeptidase and cathepsins B and D was restricted to gut tissue and to those developmental stages feeding on blood.</p> <p>Conclusion</p> <p>Overall, our results demonstrate the presence of a network of cysteine and aspartic peptidases that conceivably operates to digest host blood proteins in a concerted manner. Significantly, the peptidase components of this digestive network are orthologous to those described in other parasites, including nematodes and flatworms. Accordingly, the present data and those available for other tick species support the notion of an evolutionary conservation of a cysteine/aspartic peptidase system for digestion that includes ticks, but differs from that of insects relying on serine peptidases.</p

Secondary Atomization: Drop Breakup in a Continuous Air Jet

Understanding drop breakup will optimize aircraft engine performance, reduce agro-chemical overspray, and improve pharmaceutical tablet efficacy. Large fuel fragments in engines lead to lowered fuel economy and higher pollutant emissions, while small drops yield more agro-spray drift into surrounding residential and environmental zones. Better pharmaceutical tablets will improve drug uptake and patient comfort. Engineers and scientists are currently unable to predict the number, size, and velocity of fragments formed during important drop breakup processes. Therefore, we are required to measure these quantities. We use digital inline holography (DIH) to record three-dimensional diameter and position data for fragments formed during multi-mode breakup. DIH provides 3D images at framing rates 300 times faster than in an IMAX theater. A laser is used as the light source and a high speed camera records the breakup events to video files. A MATLAB script is used to extract the diameters and positions of all fragments in the spray. The data is sorted into bins and histograms are produced which describe the probability of observing a fragment of any particular size and speed. Results show size histograms with more than one peak, a finding in direct contradiction to the last 40 years of spray research. Multiple peaks are indicative of fragmentation processes that occur due to multiple breakup mechanisms, with the number of histogram peaks corresponding to the number of mechanisms (some combination of bag, rim, and/or stamen breakup modes). The histograms will be useful to those modeling sprays in gas turbine engines and industrial sprayers

The Complexity of Piroplasms Life Cycles

Although apicomplexan parasites of the group Piroplasmida represent commonly identified global risks to both animals and humans, detailed knowledge of their life cycles is surprisingly limited. Such a discrepancy results from incomplete literature reports, nomenclature disunity and recently, from large numbers of newly described species. This review intends to collate and summarize current knowledge with respect to piroplasm phylogeny. Moreover, it provides a comprehensive view of developmental events of Babesia, Theileria, and Cytauxzoon representative species, focusing on uniform consensus of three consecutive phases: (i) schizogony and merogony, asexual multiplication in blood cells of the vertebrate host; (ii) gamogony, sexual reproduction inside the tick midgut, later followed by invasion of kinetes into the tick internal tissues; and (iii) sporogony, asexual proliferation in tick salivary glands resulting in the formation of sporozoites. However, many fundamental differences in this general consensus occur and this review identifies variables that should be analyzed prior to further development of specific anti-piroplasm strategies, including the attractive targeting of life cycle stages of Babesia or Theileria tick vectors

CT enteroclysis and CT enterography &#8212; new approaches to assessing pathology of the small intestine

Enteroklyza i enterografia tomografii komputerowej są nowoczesnymi metodami diagnostycznymi pozwalającymi na dokładną ocenę patologii ściany jelita cienkiego z jednoczesną oceną zmian pozajelitowych i pełną oceną pozostałych narządów jamy brzusznej oraz miednicy mniejszej. Podstawowa różnica w metodyce badania między enterografią a enteroklyzą polega na odmiennym sposobie podania kontrastu, w przypadku enteroklyzy odbywa się to przez sondę założoną do pętli jelita cienkiego, a w czasie enterografii kontrast podawany jest doustnie. Rolą enteroklyzy/enterografii jest rozpoznanie chorób zapalnych jelita cienkiego, dalsze monitorowanie aktywności tych chorób oraz ocena ich powikłań, ocena jelita cienkiego w przypadku podejrzenia choroby nowotworowej tego fragmentu przewodu pokarmowego, a także identyfikacja źródła krwawienia z jelita cienkiego. Kluczową kwestią dla poprawnej interpretacji wykonanego badania jest należyte przygotowanie pacjenta &#8212; wypełnienie pętli jelitowych roztworem negatywnego środka kontrastowego oraz dobór odpowiedniej techniki badania w zależności od danych zawartych w skierowaniu. Niezmiennie ważną role odgrywa doświadczenie radiologa interpretującego badanie. Pełna ocena badania obejmuje ocenę toposcanu, obrazów poprzecznych, traktowanych zawsze jako obrazy referencyjne, oraz ocenę rekonstrukcji wielopłaszczyznowych i 3D. Szczególnie enterografia w opcji tomografii komputerowej jest bezpieczną i dobrze tolerowaną przez pacjentów metodą diagnostyczną pozwalającą na zdiagnozowanie oraz monitorowanie chorób zapalnych, nowotworów i malformacji naczyniowych jelita cienkiego.CT enteroclysis and CT enterography are modern diagnostic methods that allow a detailed assessment of the small intestine wall combined with an evaluation of extraintestinal lesions and a full examination of the remaining organs in the abdominal cavity and the pelvis. A major difference in examination methodology between enterography and enteroclysis is the different way in which a contrast medium is administered; in enteroclysis this is done through a catheter inserted into the small intestine, whereas in enterography the contrast medium is given orally. The purpose of enteroclysis/enterography is to identify inflammatory diseases of the small intestine, follow up on the progression of such diseases and assess their complications, evaluate the small intestine if this part of the gastrointestinal tract is supposed to be affected by a cancerous growth, as well as find the source of bleeding from the small intestine. A key issue for the correct interpretation that should follow the examination is to prepare the patient properly: fill intestinal loops with negative contrast medium solution and choose the right examination technique depending on the information contained in the referral. An invariably crucial role is played by the experience of the radiologist responsible for the interpretation of the examination. A full evaluation of the examination involves assessing a topographic scan, axial images, which are always treated as reference, and multiplanar and 3D reconstructions. In particular, CT enterography is a safe and well-tolerated diagnostic method that allows the diagnosing and monitoring of inflammatory diseases, neoplasia and vascular malformations of the small intestine

Natural Clerodendrum-derived tick repellent: learning from Nepali culture

Ticks attaching to ear canals of humans and animals are the cause of otoacariasis, common in rural areas of Nepal. The plant Clerodendrum viscosum is used in multiple indigenous systems of medicine by ethnic communities in the Indo-Nepali-Malaysian region. Visiting the Chitwan National Park, we learned that in indigenous medicine, flower extract of C. viscosum is utilized to treat digestive disorders and extracts from leaves as tick repellent to prevent ticks from invading or to remove them from the ear canal. The objective of our study was to provide support to indigenous medicine by characterizing the in vivo effect of leave extracts on ticks under laboratory conditions and its phytochemical composition. We collected plant parts of C. viscosum (leaves and flowers) and mango (Mangifera indica) leaves at the Chitwan National Park, previously associated with repellent activity to characterize their effect on Ixodes ricinus ticks by in vivo bioassays. A Q-ToF high-resolution analysis (HPLC-ESI-QToF) was conducted to elucidate phenolic compounds with potential repellent activity. Clerodendrum viscosum and M. indica leaf extracts had the highest tick repellent efficacy (%E = 80–100%) with significant differences when compared to C. viscosum flowers extracts (%E = 20–60%) and phosphate-buffered saline. Phytochemicals with tick repellent function as caffeic acid, fumaric acid and p-coumaric acid glucoside were identified in C. viscosum leaf extracts by HPLC-ESI-QToF, but not in non-repellent flower extracts. These results support the Nepali indigenous medicine application of C. viscosum leaf extracts to repel ticks. Additional research is needed for the development of natural and green repellent formulations to reduce the risks associated with ticks resistant to acaricides.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature.Peer reviewe

Sub-lethal radiation enhances anti-tumor immunotherapy in a transgenic mouse model of pancreatic cancer

BACKGROUND: It is not uncommon to observe circulating tumor antigen-specific T lymphocytes in cancer patients despite a lack of significant infiltration and destruction of their tumors. Thus, an important goal for tumor immunotherapy is to identify ways to modulate in vivo anti-tumor immunity to achieve clinical efficacy. We investigate this proposition in a spontaneous mouse tumor model, Rip1-Tag2. METHODS: Experimental therapies were carried out in two distinctive trial designs, intended to either intervene in the explosive growth of small tumors, or regress bulky end-stage tumors. Rip1-Tag2 mice received a single transfer of splenocytes from Tag-specific, CD4(+) T cell receptor transgenic mice, a single sub-lethal radiation, or a combination therapy in which the lymphocyte transfer was preceded by the sub-lethal radiation. Tumor burden, the extent of lymphocyte infiltration into solid tumors and host survival were used to assess the efficacy of these therapeutic approaches. RESULTS: In either intervention or regression, the transfer of Tag-specific T cells alone did not result in significant lymphocyte infiltration into solid tumors, not did it affect tumor growth or host survival. In contrast, the combination therapy resulted in significant reduction in tumor burden, increase in lymphocyte infiltration into solid tumors, and extension of survival. CONCLUSIONS: The results indicate that certain types of solid tumors may be intrinsically resistant to infiltration and destruction by tumor-specific T lymphocytes. Our data suggest that such resistance can be disrupted by sub-lethal radiation. The combinatorial approach presented here merits consideration in the design of clinical trials aimed to achieve T cell-mediated anti-tumor immunity

RNA Interference in Schistosoma mansoni Schistosomula: Selectivity, Sensitivity and Operation for Larger-Scale Screening

RNA interference (RNAi) is a technique to selectively suppress mRNA of individual genes and, consequently, their cognate proteins. RNAi using double-stranded (ds) RNA has been used to interrogate the function of mainly single genes in the flatworm, Schistosoma mansoni, one of a number of schistosome species causing schistosomiasis. In consideration of large-scale screens to identify candidate drug targets, we examined the selectivity and sensitivity (the degree of suppression) of RNAi for 11 genes produced in different tissues of the parasite: the gut, tegument (surface) and otherwise. We used the schistosomulum stage prepared from infective cercariae larvae which are accessible in large numbers and adaptable to automated screening platforms. We found that RNAi suppresses transcripts selectively, however, the sensitivity of suppression varies (40%–>75%). No obvious changes in the parasite occurred post-RNAi, including after targeting the mRNA of genes that had been computationally predicted to be essential for survival. Additionally, we defined operational parameters to facilitate large-scale RNAi, including choice of culture medium, transfection strategy to deliver dsRNA, dose- and time-dependency, and dosing limits. Finally, using fluorescent probes, we show that the developing gut allows rapid entrance of dsRNA into the parasite to initiate RNAi

Mechanical thrombectomy in acute stroke – Five years of experience in Poland

Objectives Mechanical thrombectomy (MT) is not reimbursed by the Polish public health system. We present a description of 5 years of experience with MT in acute stroke in Comprehensive Stroke Centers (CSCs) in Poland. Methods and results We retrospectively analyzed the results of a structured questionnaire from 23 out of 25 identified CSCs and 22 data sets that include 61 clinical, radiological and outcome measures. Results Most of the CSCs (74%) were founded at University Hospitals and most (65.2%) work round the clock. In 78.3% of them, the working teams are composed of neurologists and neuro-radiologists. All CSCs perform CT and angio-CT before MT. In total 586 patients were subjected to MT and data from 531 of them were analyzed. Mean time laps from stroke onset to groin puncture was 250±99min. 90.3% of the studied patients had MT within 6h from stroke onset; 59.3% of them were treated with IV rt-PA prior to MT; 15.1% had IA rt-PA during MT and 4.7% – emergent stenting of a large vessel. M1 of MCA was occluded in 47.8% of cases. The Solitaire device was used in 53% of cases. Successful recanalization (TICI2b–TICI3) was achieved in 64.6% of cases and 53.4% of patients did not experience hemorrhagic transformation. Clinical improvement on discharge was noticed in 53.7% of cases, futile recanalization – in 30.7%, mRS of 0–2 – in 31.4% and mRS of 6 in 22% of cases. Conclusion Our results can help harmonize standards for MT in Poland according to international guidelines