Role of peptidoglycan recycling enzymes AmpD and AnmK in Acinetobacter baumannii virulence features

Acinetobacter baumannii is an important causative agent of hospital acquired infections. In addition to acquired resistance to many currently-available antibiotics, it is intrinsically resistant to fosfomycin. It has previously been shown that AmpD and AnmK contribute to intrinsic fosfomycin resistance in A. baumannii due to their involvement in the peptidoglycan recycling pathway. However, the role that these two enzymes play in the fitness and virulence of A. baumannii has not been studied. The aim of this study was to characterize several virulence-related phenotypic traits in A. baumannii mutants lacking AmpD and AnmK. Specifically, cell morphology, peptidoglycan thickness, membrane permeability, growth under iron-limiting conditions, fitness, resistance to disinfectants and antimicrobial agents, twitching motility and biofilm formation of the mutant strains A. baumannii ATCC 17978 ΔampD::Kan and ΔanmK::Kan were compared to the wild type strain. Our results demonstrate that bacterial growth and fitness of both mutants were compromised, especially in the ΔampD::Kan mutant. In addition, biofilm formation was decreased by up to 69%, whereas twitching movement was reduced by about 80% in both mutants. These results demonstrate that, in addition to increased susceptibility to fosfomycin, alteration of the peptidoglycan recycling pathway affects multiple aspects related to virulence. Inhibition of these enzymes could be explored as a strategy to develop novel treatments for A. baumannii in the future. Furthermore, this study establishes a link between intrinsic fosfomycin resistance mechanisms and bacterial fitness and virulence traits.ML-S was supported by the Sara Borrell Program of the Instituto de Salud Carlos III (CD17CIII/00017), and AT was supported by the Garantıa Juvenil Program of the Comunidad ́Autónoma de Madrid (PEJ2018-004820-A -MPY 387/19), is currently supported by a FPU grant (FPU20/03261) and PhD student in Biomedical Sciences and Public Health, Universidad Nacional de Educación a Distancia (UNED), Madrid, Spain ([email protected]). MM is supported by grants from the Instituto de Salud Carlos III (MP 516/19 and MPY 380/18).S

Synthesis of slow-wave structures based on capacitive-loaded lines through aggressive space mapping (ASM)

This article is focused on the automated synthesis of slow-wave structures based on microstrip lines loaded with patch capacitors. Thanks to the presence of the shunt capacitors, the effective capacitance of the line is enhanced, and the phase velocity of the structure can be made significantly smaller than the one of the unloaded line. The target is to achieve the layout of the slow-wave structure able to provide the required slow-wave ratio, characteristic (Bloch) impedance and electrical length (i.e., the usual specifications in the design of slow-wave transmission lines). To this end, a two-step synthesis method, based on the aggressive space mapping (ASM) algorithm, is proposed for the first time. Through the first ASM algorithm, the circuit schematic providing the target specifications is determined. Then, the second ASM optimizer is used to generate the layout of the structure. To illustrate the potential of the proposed synthesis method, three application examples are successfully reported. The two-step ASM algorithm is able to provide the layout of the considered structures from the required specifications, without the need of an external aid in the process.This work has been supported by MINECO-Spain (projects TEC2010-17512 METATRANSFER, TEC2010-21520-C04-01, TEC2013-47037-C5-1-R, CONSOLIDER EMET CSD2008-00066, TEC2013-40600-R and TEC2013-49221-EXP), Generalitat de Catalunya (project 2014SGR-157), Institucio Catalana de Recerca i Estudis Avancats (who has awarded Ferran Martin) and FEDER Funds. Marco Orellana acknowledges the support of the Universidad de Costa Rica, MICITT and CONICIT to study at the Univesitat Autonoma de Barcelona.Orellana, M.; Selga, J.; Sans, M.; Rodriguez Perez, AM.; Boria Esbert, VE.; Martín Antolín, JF. (2015). Synthesis of slow-wave structures based on capacitive-loaded lines through aggressive space mapping (ASM). International Journal of RF and Microwave Computer-Aided Engineering. 25(7):629-638. https://doi.org/10.1002/mmce.20901S629638257F. Martín J. Bonache M. Durán-Sindreu J. Naqui F. Paredes G. Zamora 1 25Gorur, A. (1994). A novel coplanar slow-wave structure. IEEE Microwave and Guided Wave Letters, 4(3), 86-88. doi:10.1109/75.275589Sor, J., Qian, Y., & Itoh, T. (2001). Miniature low-loss CPW periodic structures for filter applications. IEEE Transactions on Microwave Theory and Techniques, 49(12), 2336-2341. doi:10.1109/22.971618Shau-Gang Mao, & Ming-Yi Chen. (2001). A novel periodic electromagnetic bandgap structure for finite-width conductor-backed coplanar waveguides. IEEE Microwave and Wireless Components Letters, 11(6), 261-263. doi:10.1109/7260.928932Martín, F., Falcone, F., Bonache, J., Lopetegi, T., Laso, M. A. G., & Sorolla, M. (2003). New CPW low-pass filter based on a slow wave structure. Microwave and Optical Technology Letters, 38(3), 190-193. doi:10.1002/mop.11011Garcia-Garcia, J., Bonache, J., & Martin, F. (2006). Application of Electromagnetic Bandgaps to the Design of Ultra-Wide Bandpass Filters With Good Out-of-Band Performance. IEEE Transactions on Microwave Theory and Techniques, 54(12), 4136-4140. doi:10.1109/tmtt.2006.886155Bandler, J. W., Biernacki, R. M., Shao Hua Chen, Grobelny, P. A., & Hemmers, R. H. (1994). Space mapping technique for electromagnetic optimization. IEEE Transactions on Microwave Theory and Techniques, 42(12), 2536-2544. doi:10.1109/22.339794Bandler, J. W., Biernacki, R. M., Shao Hua Chen, Hemmers, R. H., & Madsen, K. (1995). Electromagnetic optimization exploiting aggressive space mapping. IEEE Transactions on Microwave Theory and Techniques, 43(12), 2874-2882. doi:10.1109/22.475649Koziel, S., Cheng, Q., & Bandler, J. (2008). Space mapping. IEEE Microwave Magazine, 9(6), 105-122. doi:10.1109/mmm.2008.929554Koziel, S., & Bandler, J. W. (2007). Space-Mapping Optimization With Adaptive Surrogate Model. IEEE Transactions on Microwave Theory and Techniques, 55(3), 541-547. doi:10.1109/tmtt.2006.890524S. Koziel Q.S. Cheng J.W. Bandler 1995 1998Koziel, S., Bandler, J. W., & Cheng, Q. S. (2010). Robust Trust-Region Space-Mapping Algorithms for Microwave Design Optimization. IEEE Transactions on Microwave Theory and Techniques, 58(8), 2166-2174. doi:10.1109/tmtt.2010.2052666Q.S. Cheng J.W. Bandler N.K. Nikolova S. Koziel 1 4L.J. Rogla J.E. Rayas-Sanchez V.E. Boria J. Carbonell 111 114P.J. Bradley 1 4P.J. Bradley 1 17 2013J. Selga A. Rodríguez V.E. Boria F. MartínJ. Selga A. Rodríguez J. Naqui M. Durán-Sindreu V.E. Boria F. Martín 2013J. Selga M. Sans A. Rodríguez J. Bonache V. Boria F. Martín 1 4Sans, M., Selga, J., Rodriguez, A., Bonache, J., Boria, V. E., & Martin, F. (2014). Design of Planar Wideband Bandpass Filters From Specifications Using a Two-Step Aggressive Space Mapping (ASM) Optimization Algorithm. IEEE Transactions on Microwave Theory and Techniques, 62(12), 3341-3350. doi:10.1109/tmtt.2014.2365477Broyden, C. G. (1965). A class of methods for solving nonlinear simultaneous equations. Mathematics of Computation, 19(92), 577-577. doi:10.1090/s0025-5718-1965-0198670-6Marqus, R., Martn, F., & Sorolla, M. (2007). Metamaterials with Negative Parameters. doi:10.1002/978047019173

Association between feline immunodeficiency virus and Leishmania infantum infections in cats : a retrospective matched case-control study

Feline leishmaniosis caused by Leishmania infantum is often associated with feline immunodeficiency virus (FIV) infection; however, the role and clinical significance of this coinfection remain unknown. This study aimed to assess whether FIV is associated with L. infantum infection in cats from canine leishmaniosis endemic areas and to report the clinical signs and hematological alterations associated with coinfection. A retrospective matched case-control study (ratio 1:2) was conducted. Data of clinical examination and complete blood count (CBC) were selected from a cohort of 705 cats examined for epidemiological studies on feline leishmaniosis conducted between 2012 and 2019. Ninety-one FIV seropositive cases and 182 FIV seronegative control cats were selected. Matching was done according to age, sex, lifestyle and geographic provenience of case cats. Rapid ELISA devices were mainly used to detect anti-FIV antibodies. Anti- Leishmania IgG antibodies were detected by indirect-immunofluorescence test (IFAT). Leishmania DNA was searched in blood, oral and conjunctival swabs by quantitative real-time PCR. Feline immunodeficiency virus seropositive cats had no hematological abnormalities suggestive of an advanced stage of FIV infection and were statistically more frequently IFAT positive, and their risk of being L. infantum antibody positive was 2.8 greater than in the FIV seronegatives. The association of FIV seropositivity with L. infantum antibody positivity was confirmed in the univariable model of logistic regression. A multivariate model found FIV infection and L. infantum PCR positivity as predictors of a positive L. infantum IFAT result. Male outdoor cats from rural or suburban areas were at risk for FIV and L. infantum antibody positivity. Clinical signs more frequently associated with the coinfection were oral lesions, pale mucous membranes and low body condition score (BCS). This study documents that FIV seropositive cats with no hematological abnormalities suggestive of an advanced stage of FIV infection are more prone to be L. infantum seroreactive by IFAT in endemic areas. Therefore, FIV seropositive cats should be tested for L. infantum antibodies and treated for preventing sand fly bites. Pale mucous membranes, low BCS and oral lesions but no CBC abnormalities were significantly associated with the coinfection. The online version contains supplementary material available at 10.1186/s13071-022-05230-w

Neuroinflammation and Neurotransmission Mechanisms Involved in Neuropsychiatric Disorders

Some classical psychiatric disorders, such as schizophrenia, autism, major depression, bipolar and obsessive‐compulsive disorders, have been related to neuroinflammatory process, immunological abnormalities, and neurotransmission impairment beyond genetic mutations. Neuroinflammation is mostly regulated by glial cells, which respond to physiological and pathological stimuli by anti‐ and pro‐inflammatory cytokine and chemokine signaling; moreover, recent studies have indicated that glial cells also respond to the neurotransmitters. Neurotransmitters regulate many biological processes, such as cell proliferation and synaptogenesis, which contribute to the formation of functional circuits. Alterations in the neurotransmission can lead to many pathological changes that occur in brain disorders. For example, studies have shown that neuroinflammation can alter the metabolism of glutamate as well as the function of its transporters, resulting in cognitive, behavioral, and psychiatric impairments. Cytokines as IL‐1β and IL‐6 appear to have an important influence in the dopaminergic and serotoninergic neurons. These data together suggest that glial cells via cytokines and abnormal regulation of neurotransmitters can influence psychiatric disorders. The present knowledge about this issue does not allow answering whether neuroinflammation is the cause or the consequence of neurotransmission imbalance and emphasizes the importance to improve in vivo imaging methods and models to elucidate this enigma

Long-lasting insecticidal nets no longer effectively kill the highly resistant Anopheles funestus of southern Mozambique

BACKGROUND: Chemical insecticides are crucial to malaria control and elimination programmes. The frontline vector control interventions depend mainly on pyrethroids; all long-lasting insecticidal nets (LLINs) and more than 80% of indoor residual spraying (IRS) campaigns use chemicals from this class. This extensive use of pyrethroids imposes a strong selection pressure for resistance in mosquito populations, and so continuous resistance monitoring and evaluation are important. As pyrethroids have also been used for many years in the Manhica District, an area in southern Mozambique with perennial malaria transmission, an assessment of their efficacy against the local malaria vectors was conducted. METHODS: Female offspring of wild-caught Anopheles funestus s.s. females were exposed to deltamethrin, lambda-cyhalothrin and permethrin using the World Health Organization (WHO) insecticide-resistance monitoring protocols. The 3-min WHO cone bioassay was used to evaluate the effectiveness of the bed nets distributed or available for purchase in the area (Olyset, permethrin LLIN; PermaNet 2.0, deltamethrin LLIN) against An. funestus. Mosquitoes were also exposed to PermaNet 2.0 for up to 8 h in time-exposure assays. RESULTS: Resistance to pyrethroids in An. funestus s.s. was extremely high, much higher than reported in 2002 and 2009. No exposure killed more than 25.8% of the mosquitoes tested (average mortality, deltamethrin: 6.4%; lambda-cyhalothrin: 5.1%; permethrin: 19.1%). There was no significant difference in the mortality generated by 3-min exposure to any net (Olyset: 9.3% mortality, PermaNet 2.0: 6.0%, untreated: 2.0%; p = 0.2). Six hours of exposure were required to kill 50% of the An. funestus s.s. on PermaNet 2.0. CONCLUSIONS: Anopheles funestus s.s. in Manhica is extremely resistant to pyrethroids, and this area is clearly a pyrethroid-resistance hotspot. This could severely undermine vector control in this district if no appropriate countermeasures are undertaken. The National Malaria Control Programme (NMCP) of Mozambique is currently improving its resistance monitoring programme, to design and scale up new management strategies. These actions are urgently needed, as the goal of the NMCP and its partners is to reach elimination in southern Mozambique by 2020

Leishmania infantum Specific Humoral and Cellular Immune Responses in Cats and Dogs: A Comparative Cross-Sectional Study

Dogs are the main reservoir of Leishmania infantum and display different immunological patterns correlating with the progression of infection to disease. Data about feline L. infantum adaptive immune response are scant. This study aimed to compare the prevalence and immune response in cats and dogs from the same endemic area of canine leishmaniosis. Stray cats (109) and rescued dogs (59) from Córdoba (Spain) were enrolled. Data about their exposure to L. infantum were analyzed by detection of parasite DNA, measurements of Leishmania-specific interferon-γ (whole blood assay in 57 cats and 29 dogs), and antibodies (enzyme-linked immunosorbent assay and immunofluorescence antibody test). An overall L. infantum prevalence of 30.5% in dogs and 30% in cats were found according to serology and PCR tests. Prevalence was 44.8% in dogs and 35.1% in cats tested also for interferon-γ production. Dogs showed higher anti-L. infantum antibody levels compared to cats. More than one-third of cats had contact with or were infected by L. infantum and they may contribute to the endemicity of leishmaniosis in the investigated region. The immunopathogenesis of feline L. infantum infection has similarities with dogs but cats show a lower level of adaptive immune response compared to dogs

Community-informed research on malaria in pregnancy in Monrovia, Liberia: a grounded theory study

Background: Liberia is a West African country that needs substantial investment to strengthen its National Malaria Control Programme (NMCP), which was disrupted during the 2014–2016 Ebola epidemic. As elsewhere, Liberian pregnant women are especially vulnerable to malaria. Understanding prevention and treatment-seeking behaviours among the population is crucial to strategize context-specifc and women-centred actions, including locally-led malaria research, to improve women’s demand, access and use of NMCP strategies against malaria in pregnancy. Methods: In 2016, after the Ebola crisis, a qualitative inquiry was conducted in Monrovia to explore populations’ insights on the aetiology, prevention and therapeutics of malaria, as well as the community and health workers’ perceptions on the utility of malaria research for pregnant women. In-depth interviews and focus group discussions were conducted among pregnant women, traditional community representatives and hospital staf (n=38), using a feminist interpretation of grounded theory. Results: The narratives indicate that some Liberians believed in elements other than mosquito bites as causes of malaria; many had a low malaria risk perception and disliked current efective prevention methods, such as insecticide-treated nets; and some would resort to traditional medicine and spiritual care to cure malaria. Access to clinic-based malaria care for pregnant women was reportedly hindered by lack of fnancial means, by unofcial user fees requested by healthcare workers, and by male partners’ preference for traditional medicine. The participants suggested that malaria research in Liberia could help to design evidence-based education to change current malaria prevention, diagnostic and treatment-seeking attitudes, and to develop more acceptable prevention technologies. Conclusion: Poverty, insufcient education on malaria, corruption, and poor trust in healthcare establishment are structural factors that may play a greater role than local traditional beliefs in deterring Liberians from seeking, access‑ ing and using government-endorsed malaria control strategies. To increase access to and uptake of preventive and biomedical care by pregnant women, future malaria research must be informed by people’s expressed needs and constructed meanings and values on health, ill health and healthcare

Insulin and Autophagy in Neurodegeneration

Crosstalk in the pathophysiological processes underpinning metabolic diseases and neurodegenerative disorders have been the subject of extensive investigation, in which insulin signaling and autophagy impairment demonstrate to be a common factor in both conditions. Although it is still somewhat conflicting, pharmacological and genetic strategies that regulate these pathways may be a promising approach for aggregate protein clearancing and consequently the delaying of onset or progression of the disease. However, as the response due to this modulation seems to be time-dependent, finding the right regulation of autophagy may be a potential target for drug development for neurodegenerative diseases. In this way, this review focuses on the role of insulin signaling/resistance and autophagy in some neurodegenerative diseases, discussing pharmacological and non-pharmacological interventions in these diseases

Pharmacogenetic Profiling in High-Risk Soft Tissue Sarcomas Treated with Neoadjuvant Chemotherapy

Neoadjuvant chemotherapy based on anthracyclines and ifosfamide for high-risk soft tissue sarcomas (STS) of the extremities and trunk is a controversial treatment option. There are sub-stantial interindividual differences in clinical outcomes in patients treated with neoadjuvant chem-otherapy. The aim of this study was to evaluate, as biomarkers, polymorphisms in genes encoding drug-metabolizing enzymes, drug transporters, or drug targets and their association with toxicity and survival in STS patients treated with neoadjuvant chemotherapy. We analysed variants in genes involved in anthracycline metabolism (ABCB1, ABCC2, NQO1, CBR3, and SLC22A16) and in ifosfamide catabolism (ALDH1A1) in 79 treated patients. Two genes showed significant association after adjusted multivariate analysis: ABCC2 and ALDH1A1. In patients treated with anthracyclines, ABCC2 rs3740066 was associated with risk of febrile neutropenia (p = 0.031), and with decreased overall survival (OS) (p = 0.024). ABCC2 rs2273697 was associated with recurrence-free survival (RFS) (p = 0.024). In patients treated with ifosfamide, ALDH1A1 rs3764435 was associated with RFS (p = 0.046). Our pharmacogenetic study shows for the first time that variants in genes regulating the metabolism of neoadjuvant chemotherapy may be helpful to predict toxicity and survival benefit in high-risk STS treated with neoadjuvant chemotherapy. Further validation studies are needed to es-tablish their clinical utility

Clinical response to pandemic H1N1 influenza virus from a fatal and mild case in ferrets

Methods: Viral strains isolated from a patient showing mild disease-M (A/CastillaLaMancha/RR5661/2009) or from a fatal case-F (A/CastillaLaMancha/RR5911/2009), both without known comorbid conditions, were inoculated in two groups of ferrets and clinical and pathological conditions were analysed. Results: Mild to severe clinical symptoms were observed in animals from both groups. A clinical score distribution was applied in which ferrets with mild clinical signs were distributed on a non-severe group (NS) and ferrets with severe clinical signs on a severe group (S), regardless of the virus used in the infection. Animals on S showed a significant decrease in body weight compared to animals on NS at 4 to 7 days post-infection (dpi). Clinical progress correlated with histopathological findings. Concentrations of haptoglobin (Hp) and serum amyloid A (SAA) increased on both groups after 2 dpi. Clinically severe infected ferrets showed a stronger antibody response and higher viral titres after infection (p = 0.001). Conclusions: The severity in the progress of infection was independent from the virus used for infection suggesting that the host immune response was determinant in the outcome of the infection. The diversity observed in ferrets mimicked the variability found in the human population.The authors kindly thank Dr. Juan Ortín for his scientific contribution. This work was partially supported by Instituto de Salud Carlos III (Programa especial de investigación sobre la gripe pándemica GR09/0023, GR09/0040, GR09/0039), AGL2013-48923-C2-02 and CIBER de Enfermedades Infecciosa