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The cattle fever tick, Rhipicephalus (Boophilus) microplus, is the most economically important tick worldwide. Infestations with this tick can lead to direct damage and cattle mortality due to the transmission of potentially deadly pathogens. Management of this tick species has been focused on the use of synthetical acaricides; however, the emergence of acaricide resistance to single or multiple active ingredients has resulted in a need for novel acaricide compounds. Among potential avenues for the discovery of novel acaricides are plant-derived compounds. The efficacy of five organic compounds (nootkatone, Stop the Bites®, BioUD®, lavender oil, and cedarwood oil) was evaluated using larval immersion tests (LITs), repellency assays, and adult immersion tests (AITs). The results from the LITs indicate that three of the organic compounds (NootkaShield™, Stop the Bites, BioUD) led to significant mortalities at low concentrations (0.2, 0.02, and 0.08%, respectively). By comparison, lavender and cedar oil led to around 90% mortality at 10 and 1% concentrations, respectively. Similarly, NootkaShield, Stop the Bites, and BioUD had strong repellent properties with over 90% repellency at the two highest concentrations tested. Using the FAO 2004 guidelines, we evaluated the effectiveness of these organic compounds at reducing the fecundity of R. (B.) microplus and show that Nootkatone, Stop the Bites, and BioUD may significantly decrease tick populations (Drummond’s index > 90% at concentrations of 5%), highlighting their potential as alternatives to synthetic acaricides for the control of cattle fever ticks.

Herein, we present the case of a 58 y–o Caucasian man with unremarkable cardiovascular history and a strong family history for cancer. He was diagnosed with lung adenocarcinoma IVb (pleura, pleural effusion, lymph nodes), PDL1: 2% TPS and absence of actionable driver mutations on DNA and RNA NGS (only ERBB2 mutation). Started first–line chemotherapy with cisplatin–pemetrexed regimen and immunotherapy with pembrolizumab. He developed common/superficial femoral and popliteal deep vein thrombosis (DVT) and pulmonary embolism (PE). The patient started low–molecular–weight heparin (LMWH) namely enoxaparin 1 mg/kg x 2/d (weight: 65 Kg). After 1 mo, CT showed PE resolution. Due to DVT persistence and patient’s preference for oral administration, LMWH was replaced with direct oral anticoagulant (DOAC) namely edoxaban 60 mg 1 cp/d. After 2 mo, resolution of common femoral and popliteal vein thrombosis with partial recanalization of superficial femoral DVT were detected. However, he developed brain metastases treated with WB radiotherapy. We decided for edoxaban full dose long–term anticoagulant treatment with monthly cardioncological surveillance. After 6 mo since cancer diagnosis, the patient was still adherent to DOAC therapy, although in a maintenance combined regimen pemetrexed–pembrolizumab based, without both drug–drug interactions and brain bleeding. Conclusions Thromboembolism is the second leading cause of death in malignancy. In this clinical case, a man with advanced lung cancer developed extended cancer–associated thrombosis (CAT). However, the appearance of brain metastases has posed a clinical dilemma: to continue or to withdraw the anticoagulant therapy and, also, which drug to choose between LMWH and DOAC. Literature is limited on the use of DOACs in cancer patients with brain metastases. Besides, current guidelines neither advocate contraindication to DOAC, nor suggest caution as requested for gastrointestinal and genitourinary sites of cancer. We decided to proceed with edoxaban therapy, despite chemotherapy combined to immunotherapy were still ongoing and the concurrent appearance of brain metastases, with good results in efficacy, patient’s adherence and, mostly, safety. In conclusion, in regard to our clinical experience, we point out DOAC as a reliable and manageable therapeutical choice in CAT, also in active cancer patients with brain metastases receiving chemo–immunotherapy.

Massive bone loss is a serious problem in the elderly. Joint replacement with modular prostheses represents the most common reconstructive technique after oncological and non-oncological resections. Megaprostheses are broadly available, adaptable and versatile and allow early mobilization and rehabilitation. Although segmental endoprosthetic implants are now widely used and despite innovations, complications remain far high. Our purpose is to establish what happens to megaprostheses of a specific anatomical site in the long term in a population with oncologic and non-oncologic indications treated at a single center solely by a few skilled surgeons.We retrospectively reviewed our institutional database. We collected 35 patients who underwent endoprosthetic reconstruction exclusively of the proximal femur for neoplastic and non-neoplastic disease between 2008 and 2021. The minimum follow-up was 12 months. Complications were collected and classified, and also adapted to the non-oncological setting.Taking into consideration the entire population, 94% of this survived the follow-up at 6 months, subsequently 85% at 1 year and 82% at 2 years. At follow-ups after 5 years, 79% of megaprostheses showed no mechanical failure. Analyzing prosthetic survival in the two groups, this was50% at 24 months after surgery in both groups, with better survival for the oncological one.Proximal femur replacement can be a valid option in treatment of oncological and non-oncological cases. Due to the high complication rate, only selected cases should undergo this kind of surgical procedure.

Proximal femur fractures are among the most common type of trauma in elderly patients, and Euthyroid sick syndrome has already been related to fractures and trauma. The evidence of a consistent inflammatory state occurring during hip fracture, made us consider as a potential marker also the neutrophil-to-lymphocyte ratio (NLR), which is already in use to measure the prognosis and guide the therapeutic management in various conditions.A retrospective observational analysis on patients affected by proximal femur fractures was conducted. Patients were divided between affected and non-affected by Euthyroid Sick Syndrome (ESS). Standard follow up was conducted at 1, 3, 6 and 12 months.79 patients were enrolled in this study. There were 19 males and 60 females, the mean age was 83.8 ± 6.5 y.o., and 44 patients were affected by ESS. Affected patients showed higher NLR values (10.2 ± 9.4 vs. 6.9 ± 3.9; p= 0.001) and higher decrease in fT3 values in the 1st post-operative day (1.8 ± 0.4 vs. 2.2 ± 0.3; p= 0.001), higher values of PTH (97.9 ± 46.2 vs. 70.1 ± 36.2; p=0.004) and lower levels of Vitamin D (18.8 ± 7.8 vs. 23.5 ± 12.9; p= 0.04). As regards complications, we found them in 27% of patients in group A, while only in 8% in Group B, with a statistically significant difference (p= 0.03).ESS and NLR are promising prognostic markers in PFF in the elderly patients. If used together, they could help in the pre- and post-operative management of the patients.

The use of megaprosthetic implants could provide substantial advantages in elderly population affected by complex fractures. The aim of the study was to identify the patients suitable to megaprosthetic implants in the treatment of lower limbs fractures, as well as periprosthetic fractures.From January 1st, 2015, to December 31st, 2021, all patients affected by femoral fractures with severe bone loss or previous surgery failure were retrospectively reviewed. ADL, IADL, SF-12 values pre- and post-operative were recorded. Hemoglobin value, NLR, PLR were recorded pre- and peri-operatively for all patients. Complications were recorded. All patients underwent a radiological follow-up. Significance was set at p ≤ 0.05.23 patients were considered eligible, 10 males and 13 females; the mean age was 72.87 years old (± 12.33), while the mean BMI was 27.2 points (± 5.2). The mean follow-up was 2 years (± 1.4). The mean preoperative ADL and IADL scores were correlated with a positive independence of the patient, while the mean postoperative scores corresponded to a moderate-low independence. Also the mean Mental and Physical SF12 scores saw a decrease in values. NLR values were higher in the first group of patients with complications.A careful multiparametric and multidisciplinary patient selection is required to identify the suitable patient to this treatment.

Anaplasma phagocytophilum, the causative agent of human granulocytic anaplasmosis (HGA), is an obligate intracellular bacterium transmitted by the bite of black-legged ticks, Ixodes scapularis. The main host cells in vertebrates are neutrophils. However, the first site of entry is in the skin during tick feeding. Given that the initial responses within skin are a crucial determinant of disease outcome in vector-borne diseases, we used a non-biased approach to characterize the transcriptional changes that take place at the bite during I. scapularis feeding and A. phagocytophilum transmission. Experimentally infected ticks were allowed to feed for 3 days on C57BL/6J mice to allow bacterial transmission and establishment. Skin biopsies were taken from the attachment site of uninfected ticks and A. phagocytophilum-infected ticks. Skin without ticks (intact skin) was used as baseline. RNA was isolated and sequenced using next-generation sequencing (NGS). The differentially expressed genes were used to identify over-represented pathways by gene ontology (GO) and pathway enrichment (PE). Anaplasma phagocytophilum transmission resulted in the activation of interferon signaling and neutrophil chemotaxis pathways in the skin. Interestingly, it also led to the downregulation of genes encoding extracellular matrix (ECM) components, and upregulation of metalloproteinases, suggesting that A. phagocytophilum delays wound healing responses and may increase vascular permeability at the bite site.

Background Vedolizumab (VDZ) is commonly used as an off-label therapy in children with ulcerative colitis (UC) but no predictive factors of response are available. Vedolizumab binds α4β7 integrin and blocks the migration of T-lymphocytes and other cells, including eosinophils, which are involved in the UC inflammatory process. The aim of our study was to explore the predictive value of pre-therapy mucosal eosinophil count on the response to vedolizumab induction therapy in a paediatric population with UC Methods This is a retrospective, monocentric study, including children with UC treated with VDZ in the Paediatric Gastroenterology Unit of Policlinico Umberto I Hospital – Sapienza University of Rome. Histopathological data were analyzed for eosinophil counts in high power fields (hpf) in the cecum, transverse colon, sigma, and rectum prior to VDZ treatment. Clinical remission and response were evaluated at week 14 based on the PUCAI score. Results Twenty-five UC patients treated with VDZ were enrolled. All patients had failed previous anti-TNF therapy. Median baseline eosinophil count was 21 eo/hpf (5-60) in the cecum, 20 eo/hpf (6-68) in transverse colon, 44 eo/hpf (12-77) in sigma and 31 eo/hpf (9-67) in rectum. Baseline median eosinophil mucosal count in sigma was significantly higher in patients in remission (51[30-82]/ vs. 19[8.5–46] eosinophils/hpf, P23 eosinophils/hpf, with a sensitivity of 85.7% and specificity of 66.7% (Youden’s index 0.83) with an area under the curve (AUC) of 0.778 Conclusion Mucosal eosinophil abundance in sigma may predict remission to vedolizumab induction in paediatric UC. More studies are warranted to confirm these preliminary results and further investigate the role of mucosal eosinophils as a marker of therapeutic response and in the pathogenesis of UC

41 y–o woman, without cardiovascular risk factors, was diagnosed with breast cancer (pT2N1aMx, G3, ER:70%, PgR:30%, HER–2/NEU:2+, Ki67:60%). Treated (2010) with adjuvant antracyclines, taxanes and hormonal therapy. Subsequently, developed (2012) bone lesions treated with radiotherapy and hepatic metastases (2016) treated with systemic infusions of bevacizumab and taxanes. Started (2017) chemoembolization (CE), due to further liver severe progression configuring a main hepatic disease. Started LIVER ARTERIAL INFUSION CHEMOTHERAPY (LAIC) with 5fluoruracil and concomitant venous systemic chemotherapy (CT) with taxanes and bevacizumab. Developed (2020) arterial catheter–related thrombosis (ACRT) recurrent after urokinase, clopidogrel and enoxaparin, in spite of thromboprohylaxis with aspirin/clopidogrel. LAIC was discontinued and CT with taxanes and trastuzumab was administrated by systemic route. Left jugular deep vein thrombosis (DVT) occured (2021), treated for 8 months with low–molecular–weight heparin (LMWH): firstly with enoxaparin 100 IU/kg twice a day and secondly, because of lack of efficacy and patient’s request for a more manageable therapy, with parnaparin 6400 IU once a day. After 8 months the patient refused any more injectable anticoagulant drugs, asking for oral root of administration. LMWH were replaced by direct oral anticoagulant (DOAC), edoxaban 30 mg/die (weight: 43 kg). After 3 months of edoxaban a duplex ultrasound showed an almost total DVT regression; neither bleedings nor dru–drug interactions with the ongoing CT were detected. Instead, the 3 months follow–up CT–scan revealed persistence of the ACRT, despite of edoxaban. Conclusions Thromboembolism is the second leading cause of death in malignancy. Venous thromboembolic events are prevalent compared to the arterial ones. However, in active cancer hypercoagulable burden is common to both kind of thrombosis and sometimes occur simultaneously. In this case, a young woman with advanced breast cancer treated with CE and LAIC developed both DVT and ACRT. According to inefficacy and patient’s preference, we replaced LMWH with edoxaban. Arterial thrombosis was persistent although treated with several antithrombotic agents, whereas edoxaban resulted effective in the DVT regression. The take home message of this case report is that DOACs could be a feasible, efficient and safe anticoagulant choice for venous cancer related thrombosis.

Ticks are important ectoparasites that can vector multiple pathogens. The salivary glands of ticks are essential for feeding as their saliva contains many effectors with pharmaceutical properties that can diminish host immune responses and enhance pathogen transmission. One group of such effectors are microRNAs (miRNAs). miRNAs are short non-coding sequences that regulate host gene expression at the tick-host interface and within the organs of the tick. These small RNAs are transported in the tick saliva via extracellular vesicles (EVs), which serve inter-and intracellular communication. Vesicles containing miRNAs have been identified in the saliva of ticks. However, little is known about the roles and profiles of the miRNAs in tick salivary vesicles and glands. Furthermore, the study of vesicles and miRNAs in tick saliva requires tedious procedures to collect tick saliva. This protocol aims to develop and validate a method for isolating miRNAs from purified extracellular vesicles produced by ex vivo organ cultures. The materials and methodology needed to extract miRNAs from extracellular vesicles and tick salivary glands are described herein.

Ehrlichia ruminantium is a tick-borne intracellular pathogen of ruminants that causes heartwater, a disease present in Sub-saharan Africa, islands in the Indian Ocean and the Caribbean, inducing significant economic losses. At present, three avirulent strains of E. ruminantium (Gardel, Welgevonden and Senegal isolates) have been produced by a process of serial passaging in mammalian cells in vitro, but unfortunately their use as vaccines do not offer a large range of protection against other strains, possibly due to the genetic diversity present within the species. So far no genetic basis for virulence attenuation has been identified in any E. ruminantium strain that could offer targets to facilitate vaccine production. Virulence attenuated Senegal strains have been produced twice independently, and require many fewer passages to attenuate than the other strains. We compared the genomes of a virulent and attenuated Senegal strain and identified a likely attenuator gene, ntrX, a global transcription regulator and member of a two-component system that is linked to environmental sensing. This gene has an inverted partial duplicate close to the parental gene that shows evidence of gene conversion in different E. ruminantium strains. The pseudogenisation of the gene in the avirulent Senegal strain occurred by gene conversion from the duplicate to the parent, transferring a 4 bp deletion which is unique to the Senegal strain partial duplicate amongst the wild isolates. We confirmed that the ntrX gene is not expressed in the avirulent Senegal strain by RT-PCR. The inverted duplicate structure combined with the 4 bp deletion in the Senegal strain can explain both the attenuation and the faster speed of attenuation in the Senegal strain relative to other strains of E. ruminantium. Our results identify nrtX as a promising target for the generation of attenuated strains of E. ruminantium by random or directed mutagenesis that could be used for vaccine production.

PURPOSE: To determine whether preoperative endothelial cell density (ECD) and/or postoperative ECD after Descemet stripping automated endothelial keratoplasty (DSAEK) are associated with late endothelial graft failure (LEGF) in the Cornea Preservation Time Study (CPTS). DESIGN: Cohort study within multicenter randomized clinical trial. PARTICIPANTS: 1,007 individuals (1,223 study eyes), mean age 70 years, undergoing DSAEK for Fuchs’ dystrophy (94% of eyes) or pseudophakic/aphakic corneal edema (PACE) (6% of eyes) and followed for up to 5 years. METHODS: Central ECD was determined by a central image analysis reading center. Preoperative ECD was determined for 1209 eyes that did not fail and 14 eyes that experienced LEGF. ECD at 6 and 12 months after DSAEK, the change in ECD from preoperative to 6 and 12 months, surgeon-reported operative complications, and postoperative graft dislocation, were investigated for an association with LEGFs unrelated to other postoperative events. Univariable and multivariable Cox proportional hazards regression models were used to assess associations. RESULTS: The cumulative probability of LEGF was 1.3% (95% CI 0.8% to 2.4%). Median (IQR) preoperative ECDs were similar for eyes with LEGF (2523 (2367, 3161) cells/mm(2)) and eyes without failure (2727 (2508, 2973) cells/mm(2)) (p=0.34). ECD at 6 months was associated with LEGF (p

PURPOSE: To examine the association of donor, recipient, and operative factors on graft dislocation after Descemet stripping automated endothelial keratoplasty (DSAEK) in the Cornea Preservation Time Study (CPTS) as well as the effects of graft dislocation and elevated IOP on graft success and endothelial cell density (ECD) 3 years postoperatively. DESIGN: Cohort study within a multi-center, double-masked, randomized clinical trial. METHODS: 1,090 individuals (1,330 study eyes), median age 70 years, undergoing DSAEK for Fuchs endothelial corneal dystrophy (94% of eyes) or pseudophakic or aphakic corneal edema (6% of eyes). Recipient eyes receiving donor corneal tissue randomized by preservation time (PT) of 0-7 days (N=675) or 8-14 days (N=655) were monitored for early or late graft failure through 3 years. Donor, recipient, operative, and postoperative parameters were recorded including graft dislocation (GD), partial detachment, and pre- and post-operative IOP. Pre- and postoperative central donor ECD were determined by a central image analysis reading center. Proportional hazards, mixed effects, and logistic regression models estimated risk ratios and {99% confidence intervals}. RESULTS: Three independent predictive factors for GD were identified: a history of donor diabetes (odds ratio {OR}: 2.29 {1.30, 4.02}), increased pre-lamellar dissection central corneal thickness (OR: 1.13 {1.01, 1.27} per 25μ increase), and operative complications (OR: 2.97 {1.24, 7.11}). Among 104 (8%) eyes with GD, 30 (28.9%) developed primary donor or early failure and 5 (4.8%) developed late failure vs. 15 (1.2%; p

Extracellular vesicles are thought to facilitate pathogen transmission from arthropods to humans and other animals. Here, we reveal that pathogen spreading from arthropods to the mammalian host is multifaceted. Extracellular vesicles from Ixodes scapularis enable tick feeding and promote infection of the mildly virulent rickettsial agent Anaplasma phagocytophilum through the SNARE proteins Vamp33 and Synaptobrevin 2 and dendritic epidermal T cells. However, extracellular vesicles from the tick Dermacentor andersoni mitigate microbial spreading caused by the lethal pathogen Francisella tularensis. Collectively, we establish that tick extracellular vesicles foster distinct outcomes of bacterial infection and assist in vector feeding by acting on skin immunity. Thus, the biology of arthropods should be taken into consideration when developing strategies to control vector-borne diseases., Extracellular vesicles have been implicated in the transmission of pathogens from the arthropod to the human host. Here the authors show that tick-derived extracellular vesicles play a role in feeding and modulate the outcome of bacterial infection.

Borrelia burgdorferi, the causative agent of Lyme disease, traverses through vastly distinct environments between the tick vector and the multiple phases of the mammalian infection that requires genetic adaptation for the progression of pathogenesis. Borrelial gene expression is highly responsive to changes in specific environmental signals that initiate the RpoS regulon for mammalian adaptation, but the mechanism(s) for direct detection of environmental cues has yet to be identified. Secondary messenger cyclic adenosine monophosphate (cAMP) produced by adenylate cyclase is responsive to environmental signals, such as carbon source and pH, in many bacterial pathogens to promote virulence by altering gene regulation. B. burgdorferi encodes a single non-toxin class IV adenylate cyclase (bb0723, cyaB). This study investigates cyaB expression along with its influence on borrelial virulence regulation and mammalian infectivity. Expression of cyaB was specifically induced with co-incubation of mammalian host cells that was not observed with cultivated tick cells suggesting that cyaB expression is influenced by cellular factor(s) unique to mammalian cell lines. The 3′ end of cyaB also encodes a small RNA, SR0623, in the same orientation that overlaps with bb0722. The differential processing of cyaB and SR0623 transcripts may alter the ability to influence function in the form of virulence determinant regulation and infectivity. Two independent cyaB deletion B31 strains were generated in 5A4-NP1 and ML23 backgrounds and complemented with the cyaB ORF alone that truncates SR0623, cyaB with intact SR0623, or cyaB with a mutagenized full-length SR0623 to evaluate the influence on transcriptional and posttranscriptional regulation of borrelial virulence factors and infectivity. In the absence of cyaB, the expression and production of ospC was significantly reduced, while the protein levels for BosR and DbpA were substantially lower than parental strains. Infectivity studies with both independent cyaB mutants demonstrated an attenuated phenotype with reduced colonization of tissues during early disseminated infection. This work suggests that B. burgdorferi utilizes cyaB and potentially cAMP as a regulatory pathway to modulate borrelial gene expression and protein production to promote borrelial virulence and dissemination in the mammalian host.

Ticks are obligate hematophagous parasites and are important vectors of a wide variety of pathogens. These pathogens include spirochetes in the genus Borrelia that cause Lyme disease, rickettsial pathogens, and tick-borne encephalitis virus, among others. Due to their prolonged feeding period of up to two weeks, hard ticks must counteract vertebrate host defense reactions in order to survive and reproduce. To overcome host defense mechanisms, ticks have evolved a large number of pharmacologically active molecules that are secreted in their saliva, which inhibits or modulates host immune defenses and wound healing responses upon injection into the bite site. These bioactive molecules in tick saliva can create a privileged environment in the host’s skin that tick-borne pathogens take advantage of. In fact, evidence is accumulating that tick-transmitted pathogens manipulate tick saliva composition to enhance their own survival, transmission, and evasion of host defenses. We review what is known about specific and functionally characterized tick saliva molecules in the context of tick infection with the genus Borrelia, the intracellular pathogen Anaplasma phagocytophilum, and tick-borne encephalitis virus. Additionally, we review studies analyzing sialome-level responses to pathogen challenge.

Infection is a significant complication in oncological megaprostheses. The purpose of our study is to indagate the innovations and new trend about the prevention of infection in this kind of surgery. The research focused on the use of antimicrobic prophylaxis, the defensive antibacterial coating and the use of silver coated.

Ticks and tick-borne diseases are a significant economic hindrance for livestock production and a menace to public health. The expansion of tick populations into new areas, the occurrence of acaricide resistance to synthetic chemical treatments, the potentially toxic contamination of food supplies, and the difficulty of applying chemical control in wild-animal populations have created greater interest in developing new tick control alternatives. Plant compounds represent a promising avenue for the discovery of such alternatives. Several plant extracts and secondary metabolites have repellent and acaricidal effects. However, very little is known about their mode of action, and their commercialization is faced with multiple hurdles, from the determination of an adequate formulation to field validation and public availability. Further, the applicability of these compounds to control ticks in wild-animal populations is restrained by inadequate delivery systems that cannot guarantee accurate dosage delivery at the right time to the target animal populations. More work, financial support, and collaboration with regulatory authorities, research groups, and private companies are needed to overcome these obstacles. Here, we review the advancements on known plant-derived natural compounds with acaricidal potential and discuss the road ahead toward the implementation of organic control in managing ticks and tick-borne diseases.

Mutational analysis is an efficient approach to identifying microbial gene function. Until recently, lack of an effective tool for Anaplasmataceae yielding reproducible results has created an obstacle to functional genomics, because surrogate systems, e.g., ectopic gene expression and analysis in E. coli, may not provide accurate answers. We chose to focus on a method for high-throughput generation of mutants via random mutagenesis as opposed to targeted gene inactivation. In our search for a suitable mutagenesis tool, we considered attributes of the Himar1 transposase system, i.e., random insertion into AT dinucleotide sites, which are abundant in Anaplasmataceae, and lack of requirement for specific host factors. We chose the Anaplasma marginale tr promoter, and the clinically irrelevant antibiotic spectinomycin for selection, and in addition successfully implemented non-antibiotic selection using an herbicide resistance gene. These constructs function reasonably well in Anaplasma phagocytophilum harvested from human promyelocyte HL-60 cells or Ixodes scapularis tick cells. We describe protocols developed in our laboratory, and discuss what likely makes them successful. What makes Anaplasmataceae electroporation competent is unknown and manipulating electroporation conditions has not improved mutational efficiency. A concerted effort is needed to resolve remaining problems that are inherent to the obligate intracellular bacteria. Finally, using this approach, we describe the discovery and characterization of a putative secreted effector necessary for Ap survival in HL-60 cells.

The E3 ubiquitin ligase X-linked inhibitor of apoptosis (XIAP) acts as a molecular rheostat for the immune deficiency (IMD) pathway of the tick Ixodes scapularis. How XIAP activates the IMD pathway in response to microbial infection remains ill defined. Here, we identified the XIAP enzymatic substrate p47 as a positive regulator of the I. scapularis IMD network. XIAP polyubiquitylates p47 in a lysine 63-dependent manner and interacts with the p47 ubiquitin-like (UBX) module. p47 also binds to Kenny (IKKγ/NEMO), the regulatory subunit of the inhibitor of nuclear factor (NF)- κB kinase complex. Replacement of the amino acid lysine to arginine within the p47 linker region completely abrogated molecular interactions with Kenny. Furthermore, mitigation of p47 transcription levels through RNA interference in I. scapularis limited Kenny accumulation, reduced phosphorylation of IKKβ (IRD5), and impaired cleavage of the NF-κB molecule Relish. Accordingly, disruption of p47 expression increased microbial colonization by the Lyme disease spirochete Borrelia burgdorferi and the rickettsial agent Anaplasma phagocytophilum. Collectively, we highlight the importance of ticks for the elucidation of paradigms in arthropod immunology. Manipulating immune signaling cascades within I. scapularis may lead to innovative approaches to reducing the burden of tick-borne diseases.

The specific interactions of members of tick bacterial microbiota and their effects on pathogen transmission remains relatively unexplored. Here, we introduced a novel Wolbachia infection type into Ixodes scapularis tick cells and examined the antipathogenic effects on the intracellular pathogen Anaplasma phagocytophilum. An increase in A. phagocytophilum replication was observed in Wolbachia-infected tick cells. However, Wolbachia infection densities decreased when cells were serially passaged and ultimately the infection was lost. Host-cell immune response was also examined as an additional factor that could have affected A. phagocytophilum replication in Wolbachia-infected cells. In early passages post-Wolbachia infection, a decreased immune response was observed, but in later passages of cells with low Wolbachia densities, there was no change in the immune response. The results are discussed in relation to the importance of studying the interactions of the tick microbiota, the host cell, and the pathogen and the development of novel tick and tick-borne disease-control approaches.

Background Scarce data have investigated the association between pediatric Inflammatory bowel disease (IBD) and Eosinophilic esophagitis (EoE). We, therefore, aimed to describe the epidemiology and a possible peculiar phenotype and natural history of such association. Methods Case-control study based on the Italian Society for Pediatric Gastroenterology (SIGENP) national registry. All children with a concomitant diagnosis of IBD and EoE were included. Overall prevalence and incidence in 2 periods, 2009–2015 and 2016–2021, were calculated. Cases were matched with IBD only and EoE only patients in a 1:3 ratio. Phenotype and outcomes (courses of steroids, risk of complications, surgery, treatment escalation, and hospitalization) were compared between groups. Results Eleven patient (age 11,27 ± 2,83, Male 91%) with EoE-IBD out of 3090 patients with IBD only were identified, resulting in a prevalence of 0,35% and an incidence of 0,18% for the period 2009–2015 and 0,45% for the period 2016–2021. Treatment escalation rates were significantly higher in patients with IBD compared to EoE-IBD at 12 and 24-month follow-up (0 vs 30%, p = 0.04 and 9% Vs 45,5%, p = 0.03, respectively). Patients with IBD only were at a significantly higher risk of hospitalization than both EoE-IBD and EoE only patients (Log Rank p < 0.001). We found no significant differences in major outcomes related to the EoE course in EoE-IBD patients compared to EoE patients. Conclusion Overall, the incidence and prevalence of EoE in children with IBD are low, although the incidence seems to be rising in the six years. Having an EoE appears to be associated with a milder IBD disease course while having an IBD doesn’t seem to affect the natural history of EoE. More data are needed to better define the phenotype of such condition.

Levosimendan was first approved for clinic use in 2000, when authorisation was granted by Swedish regulatory authorities for the haemodynamic stabilisation of patients with acutely decompensated chronic heart failure. In the ensuing 20 years, this distinctive inodilator, which enhances cardiac contractility through calcium sensitisation and promotes vasodilatation through the opening of adenosine triphosphate-dependent potassium channels on vascular smooth muscle cells, has been approved in more than 60 jurisdictions, including most of the countries of the European Union and Latin America. Areas of clinical application have expanded considerably and now include cardiogenic shock, takotsubo cardiomyopathy, advanced heart failure, right ventricular failure and pulmonary hypertension, cardiac surgery, critical care and emergency medicine. Levosimendan is currently in active clinical evaluation in the US. Levosimendan in IV formulation is being used as a research tool in the exploration of a wide range of cardiac and non-cardiac disease states. A levosimendan oral form is at present under evaluation in the management of amyotrophic lateral sclerosis. To mark the 20 years since the advent of levosimendan in clinical use, 51 experts from 23 European countries (Austria, Belgium, Croatia, Cyprus, Czech Republic, Estonia, Finland, France, Germany, Greece, Hungary, Italy, the Netherlands, Norway, Poland, Portugal, Russia, Slovenia, Spain, Sweden, Switzerland, UK and Ukraine) contributed to this essay, which evaluates one of the relatively few drugs to have been successfully introduced into the acute heart failure arena in recent times and charts a possible development trajectory for the next 20 years.

Background: The emergence of highly active novel agents has led some to question the role of autologous haematopoietic stem-cell transplantation (HSCT) and subsequent consolidation therapy in newly diagnosed multiple myeloma. We therefore compared autologous HSCT with bortezomib–melphalan–prednisone (VMP) as intensification therapy, and bortezomib–lenalidomide–dexamethasone (VRD) consolidation therapy with no consolidation. Methods: In this randomised, open-label, phase 3 study we recruited previously untreated patients with multiple myeloma at 172 academic and community practice centres of the European Myeloma Network. Eligible patients were aged 18–65 years, had symptomatic multiple myeloma stage 1–3 according to the International Staging System (ISS), measurable disease (serum M protein >10 g/L or urine M protein >200 mg in 24 h or abnormal free light chain [FLC] ratio with involved FLC >100 mg/L, or proven plasmacytoma by biopsy), and WHO performance status grade 0–2 (grade 3 was allowed if secondary to myeloma). Patients were first randomly assigned (1:1) to receive either four 42-day cycles of bortezomib (1·3 mg/m2 administered intravenously or subcutaneously on days 1, 4, 8, 11, 22, 25, 29, and 32) combined with melphalan (9 mg/m2 administered orally on days 1–4) and prednisone (60 mg/m2 administered orally on days 1–4) or autologous HSCT after high-dose melphalan (200 mg/m2), stratified by site and ISS disease stage. In centres with a double HSCT policy, the first randomisation (1:1:1) was to VMP or single or double HSCT. Afterwards, a second randomisation assigned patients to receive two 28-day cycles of consolidation therapy with bortezomib (1·3 mg/m2 either intravenously or subcutaneously on days 1, 4, 8, and 11), lenalidomide (25 mg orally on days 1–21), and dexamethasone (20 mg orally on days 1, 2, 4, 5, 8, 9, 11, and 12) or no consolidation; both groups received lenalidomide maintenance therapy (10 mg orally on days 1–21 of a 28-day cycle). The primary outcomes were progression-free survival from the first and second randomisations, analysed in the intention-to-treat population, which included all patients who underwent each randomisation. All patients who received at least one dose of study drugs were included in the safety analyses. This study is registered with the EU Clinical Trials Register (EudraCT 2009-017903-28) and ClinicalTrials.gov (NCT01208766), and has completed recruitment. Findings: Between Feb 25, 2011, and April 3, 2014, 1503 patients were enrolled. 1197 patients were eligible for the first randomisation, of whom 702 were assigned to autologous HSCT and 495 to VMP; 877 patients who were eligible for the first randomisation underwent the second randomisation to VRD consolidation (n=449) or no consolidation (n=428). The data cutoff date for the current analysis was Nov 26, 2018. At a median follow-up of 60·3 months (IQR 52·2–67·6), median progression-free survival was significantly improved with autologous HSCT compared with VMP (56·7 months [95% CI 49·3–64·5] vs 41·9 months [37·5–46·9]; hazard ratio [HR] 0·73, 0·62–0·85; p=0·0001). For the second randomisation, the number of events of progression or death at data cutoff was lower than that preplanned for the final analysis; therefore, the results from the second protocol-specified interim analysis, when 66% of events were reached, are reported (data cutoff Jan 18, 2018). At a median follow-up of 42·1 months (IQR 32·3–49·2), consolidation therapy with VRD significantly improved median progression-free survival compared with no consolidation (58·9 months [54·0–not estimable] vs 45·5 months [39·5–58·4]; HR 0·77, 0·63–0·95; p=0·014). The most common grade ≥3 adverse events in the autologous HSCT group compared to the VMP group included neutropenia (513 [79%] of 652 patients vs 137 [29%] of 472 patients), thrombocytopenia (541 [83%] vs 74 [16%]), gastrointestinal disorders (80 [12%] vs 25 [5%]), and infections (192 [30%] vs 18 [4%]). 239 (34%) of 702 patients in the autologous HSCT group and 135 (27%) of 495 in the VMP group had at least one serious adverse event. Infection was the most common serious adverse event in each of the treatment groups (206 [56%] of 368 and 70 [37%] of 189). 38 (12%) of 311 deaths from first randomisation were likely to be treatment related: 26 (68%) in the autologous HSCT group and 12 (32%) in the VMP group, most frequently due to infections (eight [21%]), cardiac events (six [16%]), and second primary malignancies (20 [53%]). Interpretation: This study supports the use of autologous HSCT as intensification therapy and the use of consolidation therapy in patients with newly diagnosed multiple myeloma, even in the era of novel agents. The role of high-dose chemotherapy needs to be reassessed in future studies, in particular in patients with undetectable minimal residual disease after four-drug induction regimens including a monoclonal antiboby combined with an immunomodulatory agent and a proteasome inhibitor plus dexamethasone. Funding: Janssen and Celgene. © 2020 Elsevier Ltd

Disc herniation is one of most common causes of spine surgery. Because of the presence of posterior longitudinal ligaments, disc fragments often migrate into the ventral epidural space. A posterior epidural herniation of a disc fragment is a rare occurrence. We report two cases of posterior migrated disc fragments, with, radiological and clinical findings. Because of the rarity of a posterior migration of the intervertebral disc fragments, a differential diagnosis can be challenging. This painful syndrome associated with neurological lower limb deficits can be confused initially, with other posterior epidural space-occupying lesions such as tumours, abscess or hematomas. A gadolinium-enhanced MRI scan is the gold standard for a correct diagnosis. Early surgical decompression of the spine with a posterior approach remains the optimal technique in ensuring the best possible outcome for the patient.

Introduction: The microbiological safety and control of the water used in dental practice has a critical importance for avoiding cross-linked infections in the dental office. The aim of this study was to establish coxsackie virus filtration of the water applied to a dental unit. Methods: A specific water filter system was used, to verify the viral load in the outgoing water. Statistical analysis was performed using the Shapiro-Wilk test and Statview software from SAS Institute. Results: The outcome of the evaluation of the virological tests shows an excellent capability of virus filtration that attested 99.9999% in the volume analysed. A statistical difference was found in the bacterial water contamination before and after filtration. (P=0.000000). Conclusions: According to the tests, medical devices applied to a dental unit are able to filter viruses and therefore reduce risk of contamination in the dental office., Journal of Preventive Medicine and Hygiene, Vol. 61 No. 2 (2020): 2020612

Wide diaphyseal bone defects, above all those infected, encounter into Masquelet technique a suitable treatment. The two-step procedure allows the surgeon to eliminate the infected tissues and then to promote new bone formation. We analyzed the literature about the use of the induced membrane technique in osteomyelitis and the innovations recently suggested.We reviewed some of the most common web databases using the key-words: Masquelet technique, induced membrane, and osteomyelitis. 66 studies were analyzed.Comparing the Masquelet technique to other surgical procedures it shows better functional results in large bone defects due to infection. The induced membrane is like a biological chamber that protects the autograft and induces new bone formation promoting growth factors secretion. Different authors tried to improve one or more steps of the surgical procedure. Some studies focused on polymethyl methacrylate role and the possibility to use different materials instead of cement to induce the membrane. Others analyzed the autograft harvesting and placing techniques trying to reduce the amount of bone essential to fill the gap, like the RIA technique. Moreover, bone substitutes have been used, as beta-tricalcium phosphate, that showed an osteoconductive ability.The survey is not a systematic review. Nevertheless, new concepts are introduced and analyzed identifying 6 areas of interest and induced membrane technique development.

The insect immune deficiency (IMD) pathway resembles the tumour necrosis factor receptor network in mammals and senses diaminopimelic-type peptidoglycans present in Gram-negative bacteria. Whether unidentified chemical moieties activate the IMD signalling cascade remains unknown. Here, we show that infection-derived lipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG) and 1-palmitoyl-2-oleoyl diacylglycerol (PODAG) stimulate the IMD pathway of ticks. The tick IMD network protects against colonization by three distinct bacteria, that is the Lyme disease spirochete Borrelia burgdorferi and the rickettsial agents Anaplasma phagocytophilum and A. marginale. Cell signalling ensues in the absence of transmembrane peptidoglycan recognition proteins and the adaptor molecules Fas-associated protein with a death domain (FADD) and IMD. Conversely, biochemical interactions occur between x-linked inhibitor of apoptosis protein (XIAP), an E3 ubiquitin ligase, and the E2 conjugating enzyme Bendless. We propose the existence of two functionally distinct IMD networks, one in insects and another in ticks., This work was supported by the National Institutes of Health (R01 AI093653 and R01AI116523 to J.H.F.P.) and the University of Maryland, Baltimore School of Medicine. E.E.M. was a trainee under the Institutional Training Grant T32AI007540 from the National Institute of Allergy and Infectious Diseases.

Vector-borne diseases cause over 700,000 deaths annually and represent 17% of all infectious illnesses worldwide. This public health menace highlights the importance of understanding how arthropod vectors, microbes and their mammalian hosts interact. Currently, an emphasis of the scientific enterprise is at the vector–host interface where human pathogens are acquired and transmitted. At this spatial junction, arthropod effector molecules are secreted, enabling microbial pathogenesis and disease. Extracellular vesicles manipulate signaling networks by carrying proteins, lipids, carbohydrates and regulatory nucleic acids. Therefore, they are well positioned to aid in cell-to-cell communication and mediate molecular interactions. This Review briefly discusses exosome and microvesicle biogenesis, their cargo, and the role that nanovesicles play during pathogen spread, host colonization and disease pathogenesis. We then focus on the role of extracellular vesicles in dictating microbial pathogenesis and host immunity during transmission of vector-borne pathogens.

The study aimed to determine the continuous rate infusion of tramadol associated with peri- and postoperative analgesia for orthopedic surgeries in dogs, as well as cardiorespiratory and adverse effects. Thirty dogs aged 4.2±1.2 years and weighing 15.1±0.9kg were enrolled in the study, premedicated intramuscularly with acepromazine (0.04mg kg-1) and tramadol (2mg kg-1); anesthesia was induced with propofol and maintained with isoflurane in oxygen. Three infusion rates were compared, comprising three experimental groups: G2: 2.0mg kg-1 h-1; G2.5: 2.5mg kg-1 h-1; and G3: 3.0mg kg-1 h-1. Surgery was initiated 15 minutes following the start of tramadol infusion. During anesthesia, animals were monitored in predefined time points: immediately after tracheal intubation and start of inhalation anesthesia (T0); surgical incision (TSI); final suture (TFS) and end of tramadol infusion (TEI), which was maintained for at least 120 minutes and prolonged according to the duration of surgery. Postoperative analgesia was evaluated through an interval pain scoring scale and the Melbourne pain scale. The mean time of tramadol infusion was greater than 120 minutes in all groups and no differences were found among them (141±27 minutes in G2, 137±27 minutes in G2.5 and 137±30 minutes in G3). Perioperative analgesia was regarded as short and did not correlate with infusion rates. Tramadol infusion provided adequate analgesia with cardiorespiratory stability Analgesia was not dose-dependent, however, and residual postoperative effects were short-lasting, which warrants proper postoperative analgesia following tramadol infusion. Additional studies are required using higher infusion rates and standardized nociceptive stimulation in order to determine how doses influence tramadol analgesia and whe therthereis a limit to its effect in dogs. RESUMO Objetivou-se determinar a infusão de taxa contínua de tramadol associada à analgesia peri e pós-operatória para cirurgias ortopédicas em cães, além de efeitos cardiorrespiratórios e adversos. Foram utilizados 30 cães, com idade média de 4,2±1,2 anos e pesos médios de 15,1±0,9kg, pré-medicados por via intramuscular com acepromazina (0,04mg/kg) e tramadol (2mg/kg). A anestesia foi induzida com propofol e mantida com isoflurano em oxigênio. Foram comparadas três taxas de infusão, compreendendo três grupos experimentais: G2: 2,0mg/kg; G2,5: 2,5mg/kg1; e G3: 3,0mg/kg. A cirurgia começou 15 minutos após o início da infusão de tramadol. Durante a anestesia, os animais foram monitorados nos seguintes momentos: imediatamente após a intubação traqueal e o início da anestesia inalatória (T0); incisão cirúrgica (TSI); final de sutura (TFS) e final da infusão de tramadol (TEI), que foi mantida por, pelo menos, 120 minutos e prolongada de acordo com a duração da cirurgia. A analgesia pós-operatória foi avaliada por escalas de pontuação de dor, conforme a escala intervalar de avaliação de dor e a escala de contagem variável de avaliação de dor da Universidade de Melbourne, a cada uma hora. O tempo médio de infusão de tramadol foi maior que 120 minutos em todos os grupos, e não foram encontradas diferenças entre elas (141±27 minutos em G2, 137±27 minutos em G2,5 e 137±30 minutos em G3). A analgesia perioperatória foi adequada na maioria dos indivíduos e a pós-operatória foi considerada curta, não correlacionada àquelas com diferentes taxas de infusão. A infusão de tramadol nas taxas estudadas produziu analgesia adequada com estabilidade cardiorrespiratória. A analgesia não foi dose dependente, no entanto os efeitos residuais pós-operatórios foram considerados curtos, o que determina a necessidade de analgesia adequada após infusão contínua de tramadol. Estudos adicionais que utilizam taxas mais elevadas de infusão de tramadol e estimulação nociceptiva padrão são necessários para determinar em que medida as doses influenciam a analgesia de tramadol e se há um limite nos seus efeitos nos cães.

To comprehensively measure the impacts of cataract surgery on patients' activities and mental and physical health, caregivers' well-being, and household incomes in a resource-poor setting Methods: One thousand two hundred thirty-four bilaterally blind older adults in Amhara region, Ethiopia, were interviewed at baseline and 1030 (83%) re-interviewed at follow-up 1 year later. Six hundred ninety three (45%) at baseline were diagnosed with cataracts and offered free surgery, of which 484 (73%) were operated. Difference-in-difference was used to estimate impacts of surgery, using surgery-ineligible, mostly non-cataract blind as controls.For patients, surgery resulted in a 0.31 standard deviation increase in an index of social participation (p 0.001), a 30% proportional increase in ability to perform activities of daily living (p 0.001), and a 17% proportional reduction in Center for Epidemiologic Studies Depression Scale (CES-D) depression score (p 0.001). A small (6%) increase in work participation occurred among men (p = 0.093) in this elderly sample. No change occurred in individual or household food insecurity, household consumption, or assets. Caregivers' mental health improved slightly (7.3% proportional reduction in CES-D; p = 0.024). Estimates show no change in caregiver work participation or social participation; however, subjective responses regarding changes from surgery suggest that reduced caregiving time was an important benefit to households.Cataract surgery significantly improved the vision as well as mental health, social engagement, and physical functioning of older adults. However, increases in work participation were very limited, likely reflecting the advanced age of the patients (mean = 76 years); in addition, possibly, to cumulative impacts of blindness on ability to work or on household assets. Earlier surgery may lead to larger economic effects.

Enterobacteriaceae are the main etiological agents of environmental bovine mastitis and are often resistant to antimicrobials, especially β-lactam class due to the production of β-lactamases. These can be classified into several groups according to the molecular structure and substrate activity. The present study evaluated the profile of pheno-genotypic resistance to β-lactams, with emphasis on the functional detection of β-lactamases. The isolates were obtained from milk samples from farms located in the interior region of the state of Rio de Janeiro. 47.6% (20/42) of the bacterial isolates producing enzymes classified as Group 1 (AmpC type) were detected, and in disk approximation test, 30% were classified as the inducible type and 70% as not inducible. To the best of the authors’ knowledge, this is the first report on AmpC β-lactamase production in enterobacteria involved in mastitis etiology in Brazil. Key words: Raw milk, environmental mastitis, dairy cattle, antimicrobial resistance.

It is estimated that approximately one billion people are at risk of infection with obligate intracellular bacteria, but little is known about the underlying mechanisms that govern their life cycles. The difficulty in studying Chlamydia spp., Coxiella spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp. and Orientia spp. is, in part, due to their genetic intractability. Recently, genetic tools have been developed; however, optimizing the genomic manipulation of obligate intracellular bacteria remains challenging. In this Review, we describe the progress in, as well as the constraints that hinder, the systematic development of a genetic toolbox for obligate intracellular bacteria. We highlight how the use of genetically manipulated pathogens has facilitated a better understanding of microbial pathogenesis and immunity, and how the engineering of obligate intracellular bacteria could enable the discovery of novel signalling circuits in host-pathogen interactions.