Host Blood Research Articles

Human growth factor-mediated signalling through lipid rafts regulates stem cell proliferation, development and survival of Schistosoma mansoni.

Although the mechanisms by which schistosomes grow and develop in humans are poorly defined, their unique outer tegument layer, which interfaces with host blood, is considered vital to homeostasis of the parasite. Here, we investigated the importance of tegument lipid rafts to the biology of Schistosoma mansoni in the context of host-parasite interactions. We demonstrate the temporal clustering of lipid rafts in response to human epidermal growth factor (EGF) during early somule development, concomitant with the localization of anteriorly orientated EGF receptors (EGFRs) and insulin receptors, mapped using fluorescent EGF/insulin ligand. Methyl-β-cyclodextrin (MβCD)-mediated depletion of cholesterol from lipid rafts abrogated the EGFR/IR binding at the parasite surface and led to modulation of protein kinase C, extracellular signal-regulated kinase, p38 mitogen-activated protein kinase and Akt signalling pathways within the parasite. Furthermore, MβCD-mediated lipid raft disruption, and blockade of EGFRs using canertinib, profoundly reduced somule motility and survival, and attenuated stem cell proliferation and somule growth and development particularly to the fast-growing liver stage. These findings provide a novel paradigm for schistosome development and vitality in the host, driven through host-parasite interactions at the tegument, that might be exploitable for developing innovative therapeutic approaches to combat human schistosomiasis.

Microbial metabolites as modulators of host physiology.

The gut microbiota is increasingly recognised as a key player in influencing human health and changes in the gut microbiota have been strongly linked with many non-communicable conditions in humans such as type 2 diabetes, obesity and cardiovascular disease. However, characterising the molecular mechanisms that underpin these associations remains an important challenge for researchers. The gut microbiota is a complex microbial community that acts as a metabolic interface to transform ingested food (and other xenobiotics) into metabolites that are detected in the host faeces, urine and blood. Many of these metabolites are only produced by microbes and there is accumulating evidence to suggest that these microbe-specific metabolites do act as effectors to influence human physiology. For example, the gut microbiota can digest dietary complex polysaccharides (such as fibre) into short-chain fatty acids (SCFA) such as acetate, propionate and butyrate that have a pervasive role in host physiology from nutrition to immune function. In this review we will outline our current understanding of the role of some key microbial metabolites, such as SCFA, indole and bile acids, in human health. Whilst many studies linking microbial metabolites with human health are correlative we will try to highlight examples where genetic evidence is available to support a specific role for a microbial metabolite in host health and well-being.

Pristine gelatin incorporation as a strategy to enhance the biofunctionality of poly(vinyl alcohol)-based hydrogels for tissue engineering applications.

Synthetic polymers, such as poly(vinyl alcohol) (PVA), are popular biomaterials for the fabrication of hydrogels for tissue engineering and regenerative medicine (TERM) applications, as they provide excellent control over the physico-chemical properties of the hydrogel. However, their bioinert nature is known to limit cell-biomaterial interactions by hindering cell infiltration, blood vessel recruitment and potentially limiting their integration with the host tissue. Efforts in the field have therefore focused on increasing the biofunctionality of synthetic hydrogels, without limiting the advantages associated with their tailorability and controlled release capacity. The aim of this study was to investigate the suitability of pristine gelatin to enhance the biofunctionality of tyraminated PVA (PVA-Tyr) hydrogels, by promoting cell infiltration and host blood vessel recruitment for TERM applications. Pure PVA-Tyr hydrogels and PVA-Tyr hydrogels incorporated with vascular endothelial growth factor (VEGF), a well-known pro-angiogenic stimulus, were used for comparison. Incorporating increasing concentrations of VEGF (0.01-10 μg mL-1) or gelatin (0.01-5 wt%) did not influence the physical properties of PVA-Tyr hydrogels. However, their presence within the polymer network (>0.1 μg mL-1 VEGF and >0.1 wt% gelatin) promoted endothelial cell interactions with the hydrogels. The covalent binding of unmodified gelatin or VEGF to the PVA-Tyr network did not hamper their inherent bioactivity, as they both promoted angiogenesis in a chick chorioallantoic membrane (CAM) assay, performing comparably with the unbound VEGF control. When the PVA-Tyr hydrogels were implanted subcutaneously in mice, it was observed that cell infiltration into the hydrogels was possible in the absence of gelatin or VEGF at 1- or 3-weeks post-implantation, highlighting a clear difference between in vitro an in vivo cell-biomaterial interaction. Nevertheless, the presence of gelatin or VEGF was necessary to enhance blood vessel recruitment and infiltration, although no significant difference was observed between these two biological molecules. Overall, this study highlights the potential of gelatin as a standalone pro-angiogenic cue to enhance biofunctionality of synthetic hydrogels and provides promise for their use in a variety of TERM applications.

Mechanical strategies to promote vascularization for tissue engineering and regenerative medicine.

Vascularization is a major challenge in the field of tissue engineering and regenerative medicine. Mechanical factors have been demonstrated to play a fundamental role in vasculogenesis and angiogenesis and can affect the architecture of the generated vascular network. Through the regulation of mechanical factors in engineered tissues, various mechanical strategies can be used to optimize the preformed vascular network and promote its rapid integration with host vessels. Optimization of the mechanical properties of scaffolds, including controlling scaffold stiffness, increasing surface roughness and anisotropic structure, and designing interconnected, hierarchical pore structures, is beneficial for the in vitro formation of vascular networks and the ingrowth of host blood vessels. The incorporation of hollow channels into scaffolds promotes the formation of patterned vascular networks. Dynamic stretching and perfusion can facilitate the formation and maturation of preformed vascular networks in vitro. Several indirect mechanical strategies provide sustained mechanical stimulation to engineered tissues in vivo, which further promotes the vascularization of implants within the body. Additionally, stiffness gradients, anisotropic substrates and hollow channels in scaffolds, as well as external cyclic stretch, boundary constraints and dynamic flow culture, can effectively regulate the alignment of vascular networks, thereby promoting better integration of prevascularized engineered tissues with host blood vessels. This review summarizes the influence and contribution of both scaffold-based and external stimulus-based mechanical strategies for vascularization in tissue engineering and elucidates the underlying mechanisms involved.

Forensic analysis of mosquito blood meal digestion process and the impact of heroin opiate: determination of the post-feeding interval as a PMI estimation.

Females of some mosquito species are anthropophilic, as they feed on human blood to support egg production and, hence, are forensically valuable if found at a crime scene. The present study investigated the blood meal digestion process in Culex pipiens L. both with and without heroin and proposed a method for estimating the post-feeding interval (PFI). Mosquitoes were fed on a control mouse, a heroin-injected mouse, or in vitro heroin-treated mouse blood. The blood meal digestion was then investigated at different hours post-feeding. Data showed that the blood meal size ingested by control mosquitoes was 0.681 ± 0.04 mg/mosquito and was completely digested within 45 h post-feeding. An estimation of the PFI was proposed in terms of the rate of hemoglobin (Hb) digestion. The blood meal size of the mosquitoes fed on the in vitro heroin-treated blood and the heroin-injected mouse was 0.96 ± 0.06 and 0.79 ± 0.01 mg/mosquito and was completely digested within 50 and 55 h post-feeding, respectively. The digestion of Hb started similarly in all experimental mosquitoes until 10 h post-feeding, after which it significantly decreased in heroin-treated blood meals compared with the control ones. This may suggest that heroin impacted the digestion process, as it took an extra 5-10 h to complete. These findings could be valuable in the forensic context since an estimation of PFI is proposed as a potential estimation of the postmortem interval (PMI). However, care should be taken as heroin in the host blood has significantly impacted the overall digestion process and, hence, may bias the PFI/PMI estimation.

Guardians of the Herd: Molecular Surveillance of Tick Vectors Uncovers Theileriosis Perils in Large Ruminants.

Theileria sp. (Piroplasmida: Theileriidae) is one of the most widely known infections transmitted by hard ticks (Acari: Ixodidae) and has been linked to significant economic losses across the globe. The study's main emphasis was theileriosis, a disease that is common in Pakistan and has an incidence ranging from 0.6% to 33%. Through DNA screening of the vector ticks and host blood, this study sought to determine the risk of tick-borne theileriosis in populations of buffalos (Bubalus bubalis) and cattle (Bos indicus) in Toba Tek Singh district of Punjab, Pakistan. Identified tick species include Hyalomma anatolicum (35.4%), Rhipicephalus (Boophilus) microplus (30.2%), and R. sanguineus (25%). Tick specimens were collected from animals and their respective microenvironments. PCR assays targeting Theileria annulata were used to investigate the infection in the DNA extracted from the collected blood samples from large ruminants and salivary glands (SGs) of the Hyalomma ticks. The 18S rRNA of T. annulata was amplified using specific primers. Positive T. annulata amplicons were sequenced and verified using BLAST analysis. Overall, 50% of SGs contained T. annulate DNA. Female ticks, and those collected from cattle and from riverine environments had significantly higher (p < 0.05) rates of Theileria infection in their acini. Overall prevalence of Theileria infection was 35.9% in blood collected from large ruminants. Cattle had a substantially greater frequency of bovine theileriosis (43.2%) than buffalos (28.7%). Age and sex of large ruminants were significantly positively associated (p < 0.05) with Theileria infection. Furthermore, compared to non-riverine cattle (35%) and buffalo (19.5%), riverine cattle (52.2%) and buffalo (36.2%) showed a considerably higher prevalence. The results of this study, which is the first in Pakistan to examine the blood of large ruminants and vectorial function of Ixodid ticks in the transmission of T. annulata along with associated risk factors, offer an important insight for risk assessment of Theileria infection in livestock using vectorial infectivity.

Enolase Inhibitors as Early Lead Therapeutics against Trypanosoma brucei.

Glucose metabolism is critical for the African trypanosome, Trypanosoma brucei, serving as the lone source of ATP production for the bloodstream form (BSF) parasite in the glucose-rich environment of the host blood. Recently, phosphonate inhibitors of human enolase (ENO), the enzyme responsible for the interconversion of 2-phosphoglycerate (2-PG) to phosphoenolpyruvate (PEP) in glycolysis or PEP to 2-PG in gluconeogenesis, have been developed for the treatment of glioblastoma multiforme (GBM). Here, we have tested these agents against T. brucei ENO (TbENO) and found the compounds to be potent enzyme inhibitors and trypanocides. For example, (1-hydroxy-2-oxopyrrolidin-3-yl) phosphonic acid (deoxy-SF2312) was a potent enzyme inhibitor (IC50 value of 0.60 ± 0.23 µM), while a six-membered ring-bearing phosphonate, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX), was less potent (IC50 value of 2.1 ± 1.1 µM). An analog with a larger seven-membered ring, (1-hydroxy-2-oxoazepan-3-yl) phosphonic acid (HEPTA), was not active. Molecular docking simulations revealed that deoxy-SF2312 and HEX had binding affinities of -6.8 and -7.5 kcal/mol, respectively, while the larger HEPTA did not bind as well, with a binding of affinity of -4.8 kcal/mol. None of these compounds were toxic to BSF parasites; however, modification of enzyme-active phosphonates through the addition of pivaloyloxymethyl (POM) groups improved activity against T. brucei, with POM-modified (1,5-dihydroxy-2-oxopyrrolidin-3-yl) phosphonic acid (POMSF) and POMHEX having EC50 values of 0.45 ± 0.10 and 0.61 ± 0.08 µM, respectively. These findings suggest that HEX is a promising lead against T. brucei and that further development of prodrug HEX analogs is warranted.

Insight Into the Dynamics of the Ixodes ricinus Nymphal Midgut Proteome

Ticks are ectoparasites that feed on blood and have an impressive ability to consume and process enormous amounts of host blood, allowing extremely long periods of starvation between blood meals. The central role in the parasitic lifestyle of ticks is played by the midgut. This organ efficiently stores and digests ingested blood and serves as the primary interface for the transmission of tick-borne pathogens. In this study, we used a label-free quantitative approach to perform a novel dynamic proteomic analysis of the midgut of Ixodesricinus nymphs, covering their development from unfed to pre-molt stages. We identified 1534 I.ricinus-specific proteins with a relatively low proportion of host proteins. This proteome dataset, which was carefully examined by manual scrutiny, allowed precise annotation of proteins important for blood meal processing and their dynamic changes during nymphal ontogeny. We focused on midgut molecules related to lipid hydrolysis, storage, and transport, opening a yet unexplored avenue for studying lipid metabolism in ticks. Further dynamic profiling of the tick's multi-enzyme digestive network, protease inhibitors, enzymes involved in redox homeostasis and detoxification, antimicrobial peptides, and proteins responsible for midgut colonization by Borrelia spirochetes promises to uncover new targets for targeting tick nymphs, the most critical life stage for transmission the pathogens that cause tick-borne diseases.

SAT041 18F-FDG PET/CT Mapping Of Functional Microbial Niches To Understand Host Glucose Regulation After BCG Vaccinations

Abstract Disclosure: H. Dias: None. J.F. Fu: None. T.G. Luck: None. G.E. Wolfe: None. E. Hostetter: None. N.C. Ng: None. H. Zheng: None. W. Kühtreiber: None. J.C. Price: None. C. Catana: None. D.L. Faustman: None. Diverse microbiomes in humans are known to change host energy metabolism. When the bacillus Calmette-Guérin (BCG) vaccine, which consists of live but avirulent Mycobacterium bovis, is introduced as experimental therapy into human hosts with underlying type 1 diabetes (T1D), the BCG bacillus has been shown to gradually shift energy metabolism in host blood lymphoid cells from oxidative phosphorylation to aerobic glycolysis. Aerobic glycolysis draws more glucose out of the blood to fuel intracellular metabolism, and it may underlie the BCG bacillus’ therapeutic benefit: the systemic and long-term reduction of excess glucose from the blood of TID patients. The organ-specific niches where the BCG bacillus similarly alters metabolism and establishes persistent residence are not known. In a human clinical trial, we mapped the organ(s) niches for BCG-induced shift to aerobic glycolysis using fluorine-18 fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) and x-ray computed tomography (CT). This technique allowed us to reveal quantitatively, by Standardized Uptake Value Ratios, the organs with heightened glucose uptake in T1D participants (n=6) over a 2-year time-period after BCG vaccination versus before vaccination, as well as confirm earlier work that BCG vaccination gradually lowers blood sugar levels without any contribution from endogenous insulin. We also tested BALB/c mice (n=17) for the direct presence of BCG colonies in particular organ niches before and after vaccination. Results from both human and murine studies of the BCG microorganism concurred that the major anatomic site of functional metabolic change and residence is the spleen. The BCG bacillus also transiently mapped to the bone marrow, liver, circulating lymphocytes as in the descending aorta, and the lungs. Findings from human and murine studies support the spleen as the niche for the BCG vaccine’s functional improvement of metabolism. The spleen is significant because it is a lymphoid organ massive enough to explain BCG’s systemic benefit of lowering blood sugar to near normal levels in T1D. Presentation: Saturday, June 17, 2023

Acetobacter and lactobacillus alleviate the symptom of insulin resistance by blocking the JNK-JAK/STAT pathway in Drosophila melanogaster

The dysregulation of intestinal microbiota is well-known to be one of the main causes of insulin resistance in both vertebrates and invertebrates. Specially, the acetobacter and lactobacillus have been identified as potentially capable of alleviating insulin resistance. However, the molecular mechanism underlying this effect requires further elucidation. In this study, we employed Drosophila melanogaster (fruit fly) as a model organism to delineate how intestinal microbiota disrupts the host intestinal signaling pathway, contributing to insulin resistance. Our findings demonstrate that a long-term high-sugar diet lead to a reduction in the general diversity of intestinal microbiota in flies, as well as a marked decrease in the abundances of acetobacter and lactobacillus. Furthermore, we observed that symptoms of insulin resistance were alleviated by feeding flies with acetobacter or lactobacillus, indicating that these microorganisms play an essential role in maintaining blood sugar homeostasis in flies. Conversely, when all intestinal microbiota was removed, flies show severe symptoms of insulin resistance, confirming that the critical role of intestinal microbiota in maintaining host blood sugar homeostasis. Our studies suggested that the intestinal but not fat body JNK pathway mediates the communication of intestinal microbiota and host insulin pathway. In flies, downregulation of JNK activity alleviates symptoms of insulin resistance by decreasing the activity of the JAK/STAT pathway. However, this offsets the therapeutic effects of supplying flies with acetobacter or lactobacillus, suggesting that the therapeutic function of these microorganisms is based on their interaction with JNK-JAK/STAT axis. Taken together, our study reveals that acetobacter and lactobacillus alleviate insulin resistance symptoms in a JNK-JAK/STAT pathway-dependent manner, indicating the therapeutic potential of probiotic supplementation and regulation of the activities of JNK-JAK/STAT pathway for diabetes control.

Therapeutic Management of Canine Dirofilariasis in Nagaland, India

Dirofilariasis is one of the most clinically important diseases in dogs. It is also called as heart-worm disease, which is a non-contagious parasitic disease caused by filarial or small thread-like worms, Dirofilaria immitis, and D. repens of the family Onchocercidae (Nelson et al., 2014; Adebayo et al., 2022). It is an arthropod-borne disease. The mosquitoes belonging to the genera Culex, Aedes, Anopheles, and Mansonia are principally responsible for the transmission of this disease (Borthakur et al., 2016). Transmission of the parasite occurs when a potential vector bites dogs during a blood meal. It takes about 6-7 months for the third-stage larvae (L3) to become adult parasites after entering the blood vessels of the hosts (Koutsu et al., 2022). After maturation, the adult worms reproduce and produce microfilariae (pre-L1), which are released in the host blood vessels and taken up by a feeding mosquito (Thilakarathne et al., 2023), and completed the lifecycle of the parasite.

Obesity is the main driver of altered gut microbiome functions in the metabolically unhealthy

ABSTRACT Obesity (OB) and cardiometabolic disease are major public health issues linked to changes in the gut microbiome. OB and poor cardiometabolic health status (CHS) are often comorbid, which hinders efforts to identify components of the microbiome uniquely linked to either one. Here, we used a deeply phenotyped cohort of 408 adults from Colombia, including subjects with OB, unhealthy CHS, or both, to validate previously reported features of gut microbiome function and diversity independently correlated with OB or CHS using fecal metagenomes. OB was defined by body mass index, waist circumference, and body fat; CHS as healthy or unhealthy according to blood biochemistry and anthropometric data. We found that OB, more so than metabolic status, drove associations with gut microbiome structure and functions. The microbiome of obese individuals with and without co-existing unhealthy CHS was characterized by reduced metagenomic diversity, reduced fermentative potential and elevated capacity to respond to oxidative stress and produce bacterial antigens. Disease-linked features were correlated with increased host blood pressure and inflammatory markers, and were mainly contributed by members of the family Enterobacteriaceae. Our results link OB with a microbiome able to tolerate an inflammatory and oxygenated gut state, and suggest that OB is the main driver of microbiome functional differences when poor CHS is a comorbidity.

The neuro-exocrine secretion: A new type of gland in tapeworms?

The phenomenon of exocrine secretion via nervous cells into the host tissue has been discovered in cestodes. In five cestode species of different orders specialized “cup-shaped” free nerve endings located in the tegument have been found. Their ultrastructure is characterized by the presence of a septate junction, a thin support ring and neurosecretory vesicles 90–110 nm in diameter, which are secreted onto the surface of the tegument through a thin pore. The phenomenon is referred to in this article as the neuro-exocrine secretion. We observed a direct relationship between neurosecretory processes in the deep subtegument and free endings in a series of ultrathin sections in two species. The peripheral neurosecretory neurons of species studied are characterized by similar ultrastructural features: size and location; diameter of neurosecretory granules; absence of microtubules and mitochondria in the neurites. The size of neurosecretory granules has been found to decrease from perikaryon towards neurosecretory terminals that lead to the tegument. In two species, we examined the neurosecretion during incubation in the host's blood serum. Depending on the time of incubation we have shown the changes a) in the diameter of the cup-shaped endings, b) in the number of secretory vesicles in the endings; c) changes in number and diameter of neurosecretory vesicles in the processes of neurosecretory neurons in the subtegument. The detected changes differ in D.dendriticus and L.interrupta and, taken together, indirectly confirm the secretory specialization of the cup-shaped endings. Supposed targets for the neurosecretory neurons in the studied cestodes are the following: (a) eccrine frontal gland ducts, especially their terminal regions involved in the release of secretory products; (b) longitudinal and circular muscles in the subtegument region; (c) the basal membrane of the tegument. Besides the discovered secretion vesicles through the cup-shaped terminals, we observed vacuoles derived from the basal membrane of the tegument containing extracellular substances released into the host tissue. Their possible role in the release of neurosecretory substances is discussed. Considering the data acquired via immunocytochemical methods, an assumption about involvement of FMRFamide-like related peptides (FaRPs) in the neuro-exocrine secretion is proposed. Possible functions of the neuro-exocrine secretion are discussed in the context of host-parasite interactions.

Occurrence, Activity and Control Options Against Biting Midges (Diptera: Culicoides) in Horses

Abstract Biting midges are insects which annoy horses as they cause discomfort and by the sucking of blood, they cause painful lesions that, in certain cases, can trigger acute allergic reactions. They also act as potential vectors of many pathogens of bacterial, viral and parasitic origin. The aim of this study was to investigate seasonal dynamics, abundance and species composition of biting midges, and implementing protection options at the Equestrian centre of the University of Veterinary Medicine and Pharmacy (UVMP) in Košice in the years 2021 and 2022. During this time period (of 2021 and 2022), we captured 4797 biting midges. Our results showed that in 2021 the most frequently captured species were C. obsoletus/C. scoticus (prevalence 86.6 %), followed by C. punctatus and C. pulicaris (prevalence 5.37 % and 3.86 %, resp.). In addition, C. lupicaris, C. newsteadi, C. furcillatus, C. festivipennis, C. slovacus, C. tauricus, C. clastrieri, C. dewulfi, C. fagineus and C. circumscriptus were identified as the minority species. In this year, the highest abundance of these insects (1758 biting midges) was observed at the end of June. In 2022, we detected a clear dominance of C. obsoletus/C. scoticus (prevalence 97.0 %) and the highest number of biting midges (655) was captured in mid-May). In this year, the minority species included, C. pulicaris, C. punctatus and C. lupicaris. The identification of the host blood showed that man (Homo sapiens) was the dominant host, namely of 80 % of the examined biting midges of species C. obsoletus/C. scoticus, C. dewulfi, C. punctatus and C. festivipennis. In one female of species C. fagineus we identified the host blood from horses (Equus caballusi). In 2022, we detected the blood of a domestic rabbit (Oryctolagus cuniculus) in the species of C. obsoletus/C. scoticus.

Modeling human anti-pig xenoimmune responses in a pig artery tissue grafted humanized mouse model.

Blood vessels that contain endothelial cells (ECs) on the surface are in direct contact with host blood and are the first target of xenograft rejection. Currently, our understanding of human anti-pig vessel immune responses is primarily based on in vitro assays using pig ECs. Therefore, it is necessary to develop an animal model that permits in vivo study of human immunological rejection of pig vessels. Pig artery tissues (PAT) were transplanted into human immune system (HIS) mice or immunodeficient NSG mice (as controls). Intragraft human immune cell infiltration and antibody deposition were quantified using histology and immunohistochemistry. Donor antigen-specific immune responses were quantified using a mixed lymphocyte reaction and a complement-dependent killing assay. Pig CD31+ ECs were detected and increased 2-fold from weeks 3 to 5 in PAT xenografts from immunodeficient NSG mice. However, compared with NSG mice, PAT xenografts in HIS mice had significantly lower numbers of porcine CD31+ ECs and showed a marked reduction from week 3 to week 5. PAT xenograft rejection in HIS mice is associated with intensive infiltration of human immune cells, deposition of human IgM and IgG antibodies, and the formation of a tertiary lymphoid structure. Robust donor pig antigen-specific human T cells and antibody responses were detected in PAT-transplanted HIS mice. We have developed a humanized mouse model to evaluate human anti-pig xenoimmune responses by PAT transplantation in vivo. This model is expected to facilitate the refinement of pig gene-editing strategies (the expression on EC surface) and the testing of local immunosuppressive strategies for clinical pig organ xenotransplantation.

Abstract 086: Examining The Role Of Bacterial Extracellular Vesicles In Host Blood Pressure Regulation

We have demonstrated that disruption of the normal gut microbiota, termed gut dysbiosis, is an underlying cause of hypertension (HT). However, the mechanisms by which the microbiota influences host blood pressure (BP) are largely unknown. We hypothesized that extracellular vesicles (EVs) released by gut microbiota gain access to the circulation, deliver cargo to host tissues involved in BP regulation, inducing inflammation and ultimately HT. Bacterial EVs (bEVs) were isolated from cecal content of spontaneously hypertensive stroke prone rats (SHRSP) or Wistar-Kyoto rats (WKY) by size exclusion chromatography. BEV cargo including small RNAs, LPS, and 16s rRNA, was assessed. To directly examine the role of bEVs on BP, purified WKY and SHRP bEVs were transplanted to recipient rats of the same and opposite strain by oral gavage (1.5x10 10 EVs/rat) every other day for 4 weeks. The inflammatory status of ileum and brain were assessed by flow cytometry. Global analysis of small RNA cargo of bEVs was found to be significantly different between WKY and SHRSP (n=6, p=0.001). BEVs from SHRSP contained significantly greater LPS as compared to those from WKY (49 vs 38 EU/ml, n=6, p=0.01). EVs isolated from plasma of both strains contained bacterial 16s rRNA, demonstrating bEVs gain access to host circulation. Finally, as compared to WKY rats receiving WKY bEVs, WKY rats transplanted with SHRSP bEVs had 1) significant elevations in BP at 4 weeks (176 mmHg vs 150 mmHg, n=6, p&lt;0.01), 2) significant reduction of T regulatory cells in ileum (49% vs 58%, n=6, p&lt;0.05) and brain (9% vs 18%, n=6, p&lt;0.05), and 3) significant increases in macrophages in ileum (39% vs 6%, n=6, p&lt;0.001) and microglia in brain (24% vs 8%, n=6, p&lt;0.05). SHRSP rats transplanted with WKY bEVs had significantly lower BP at 4 weeks (160 mmHg vs 175 mmHg, n=6, p&lt;0.05), reduced macrophages in ileum (5% vs 42%, n=6, p&lt;0.001) and microglia in brain (5% vs 29%, n=6, p&lt;0.01), as compared to SHRSP receiving SHRSP bEVs. In conclusion, BEVs originating from the dysbiotic gut of hypertensive SHRSP rats carry distinct cargo, reach the host circulation, and are involved in host gut- and neuroinflammation and HT. These studies identify bEVs as a previously unrecognized mechanism for microbiota-host interaction involved in BP regulation.

C-reactive protein binds to short phosphoglycan repeats of Leishmania secreted proteophosphoglycans and activates complement.

Human C-reactive protein (CRP) binds to lipophosphoglycan (LPG), a virulence factor of Leishmania spp., through the repeating phosphodisaccharide region. We report here that both major components of promastigote secretory gel (PSG), the filamentous proteophosphoglycan (fPPG) and the secreted acid phosphatase (ScAP), are also ligands. CRP binding was mainly associated with the flagellar pocket when LPG deficient Leishmania mexicana parasites were examined by fluorescent microscopy, consistent with binding to secreted material. ScAP is a major ligand in purified fPPG from parasite culture as demonstrated by much reduced binding to a ScAP deficient mutant fPPG in plate binding assays and ligand blotting. Nevertheless, in sandfly derived PSG fPPG is a major component and the major CRP binding component. Previously we showed high avidity of CRP for LPG ligand required multiple disaccharide repeats. ScAP and fPPG only have short repeats but they retain high avidity for CRP revealed by surface plasmon resonance because they are found in multiple copies on the phosphoglycan. The fPPG from many species such as L. donovani and L. mexicana bound CRP strongly but L. tropica and L. amazonensis had low amounts of binding. The extent of side chain substitution of [-PO4-6Galβ1-4Manα1-] disaccharides correlates inversely with binding of CRP. The ligand for the CRP on different species all had similar binding avidity as the half maximal binding concentration was similar. Since the PSG is injected with the parasites into host blood pools and phosphoglycans (PG) are known to deplete complement, we showed that CRP makes a significant contribution to the activation of complement by PSG using serum from naive donors.

Thymic stromal lymphopoietin: evaluating its role as a new marker related to presence of psoriasis vulgaris and its severity

Background Thymic stromal lymphopoietin (TSLP), a proallergic cytokine, and T cell-derived CD40 ligand (CD40L) may collaborate to increase the production of IL-23 in psoriasis patients. One key cytokine, IL-23, is responsible for the unwarranted immune response in psoriasis sufferers. Aim The current study aims to shed light on the possible role of TSLP as a novel biomarker related to presence of psoriasis vulgaris lesions and its severe forms with exclusion of people with atopic dermatitis, allergic rhinitis, rheumatoid arthritis, graft versus host disease (GVHD), blood diseases and patient on anticoagulant therapy as well as pregnant and lactating females. Patients and methods 40 subjects with psoriasis vulgaris participated in the current case-controlled research, whereas 40 healthy volunteers of similar age and gender served as the control group. The degree and extent of the illness were evaluated utilizing the psoriasis area and severity index (PASI) score. Serum was separated after blood samples from the venous blood of the subjects and control participants were taken. As soon as possible, the serum samples were frozen at −20°C. The Sandwich Enzyme-Linked Immunosorbant Assay (ELISA) was utilized to quantify serum TSLP. Results The case group’s serum TSLP levels rose statistically substantially more than those of the control group. In the cases group, there was a statistically strong positive relation between serum TSLP levels and PASI scores (P: &lt; 0.001). There is a statistically strong positive relation between serum TSLP and patients’ age and illness duration, a statistically substantial rise in blood TSLP values in psoriatic arthritis patients and smokers. Conclusion Patients with psoriasis have higher serum TSLP levels, which are proportional to the disease’s severity.

Effects of antennal segments defects on blood-sucking behavior in Aedes albopictus.

After mating, female mosquitoes need a blood meal to promote the reproductive process. When mosquitoes bite infected people and animals, they become infected with germs such as viruses and parasites. Mosquitoes rely on many cues for host selection and localization, among which the trace chemical cues emitted by the host into the environment are considered to be the most important, and the sense of smell is the main way to perceive these trace chemical cues. However, the current understanding of the olfactory mechanism is not enough to meet the needs of mosquito control. Unlike previous studies that focused on the olfactory receptor recognition spectrum to reveal the olfactory mechanism of mosquito host localization. In this paper, based on the observation that mosquitoes with incomplete antennae still can locate the host and complete blood feeding in the laboratory, we proposed that there may be some protection or compensation mechanism in the 13 segments of antennae flagella, and only when the antennae are missing to a certain threshold will it affect the mosquito's ability to locate the host. Through rational-designed behavioral experiments, we found that the 6th and 7th flagellomeres on the Aedes albopictus antenna are important in the olfactory detection of host searching. This study preliminarily screened antennal segments important for host localization of Ae. albopictus, and provided a reference for subsequent cell biology and molecular biology studies on these segments. Meanwhile, the morphology and distribution of sensilla on each antenna flagellomere were also analyzed and discussed in this paper.

The bioengineering of perfusable endocrine tissue with anastomosable blood vessels

Organ transplantation is a definitive treatment for endocrine disorders, but donor shortages limit the use of this technique. The development of regenerative therapies would revolutionize the treatment of endocrine disorders. As is the case for harvested organs, the ideal bioengineered graft would comprise vascularized endocrine tissue, contain blood vessels that could be anastomosed to host vessels, have stable blood flow, and be suitable for transplantation into various sites. Here, we describe a transplantable endocrine tissue graft that was fabricated by ex vivo perfusion of tricultured cell sheets (islet β-cells, vascular endothelial cells (vECs), and mesenchymal stem cells (MSCs)) on a vascularized tissue flap of in vivo origin. The present study has three key findings. First, mild hypothermic conditions enhanced the success of ex vivo perfusion culture. Specifically, graft construction failed at 37 °C but succeeded at 32 °C (mild hypothermia), and endocrine tissue fabricated under mild hypothermia contained aggregations of islet β-cells surrounded by dense vascular networks. Second, the construction of transplantable endocrine tissue by ex vivo perfusion culture was better achieved using a vascular flap (VF) than a muscle flap. Third, the endocrine tissue construct generated using a VF could be transplanted into the rat by anastomosis of the graft artery and vein to host blood vessels, and the graft secreted insulin into the host’s circulatory system for at least two weeks after transplantation. Endocrine tissues bioengineered using these techniques potentially could be used as novel endocrine therapies.