Culex quinquefasciatus Holobiont: A Fungal Metagenomic Approach

Microorganisms associated with mosquitoes have fundamental roles, not only in their nutrition, but also in physiological and immunological processes, and in their adaptation to the environment as well. Studies on mosquito hologenomes have increased significantly during the last years, achieving important advances in the characterization of the “core bacteriome” of some species of health importance. However, the fungal mycobiome has not been exhaustively researched, especially throughout the life cycle of some hematophagous mosquito species. In this work, the diversity and composition of fungal communities in different developmental stages, sexes, and adult nutrition of Culex quinquefasciatus reared on laboratory conditions were characterized, using internal transcribed spacer high throughput amplicon sequencing. Larvae presented a higher fungal richness, while sucrose-fed males and females showed a similar diversity between them. Blood-fed females presented few operational taxonomic units with an even distribution. Results are consistent with the reduction of larval microbiota after molting, observed for the bacterial microbiome in other mosquito species. The filamentous Ascomycota Penicillium polonicum and Cladosporium sp. were present in all stages of the mosquitoes; in addition, the presence of yeasts in the insects or their subsequent colonization associated with their diet is also discussed. These results suggest that some species of fungi could be essential for the nutrition and development of mosquitoes throughout their life cycle.Fil: Flores, Guillermo Alejandro Máximo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología; ArgentinaFil: Lopez, Rocio de la Paz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología; ArgentinaFil: Cerrudo, Carolina Susana. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Consolo, Verónica Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología; ArgentinaFil: Berón, Corina Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología; Argentin

Antimutageno djelovanje vitamina C na oksidacijske promjene uzrokovane kinolonima

Quinolones are broad-spectrum antibiotics effective against both Gram-positive and Gram-negative bacteria. Reactive oxygen species (ROS) generated by quinolones may damage cell structures and could be a risk to health. The use of vitamin C to reduce such risks may have the opposite effects: vitamin C in the presence of divalent metal ions can induce the Fenton reaction, leading to hydroxyl radical (HO˙) generation and oxidative damage. The purpose of this study is to evaluate the antioxidant and prooxidant properties of vitamin C by measuring its effects on both lipid peroxidation and mutagenesis induced by quinolones nalidixic acid (NLX) or norfloxacin (NOR) in Salmonella typhimurium TA102. Mutagenicity was evaluated by the Ames test and the results were expressed as (histidine+ revertants/ng of quinolone), while lipoperoxidation was measured as thiobarbituric acid reactive substances (μmol malondialdehyde/(mL·h)). The effects of different concentrations of nalidixic acid (10–1000 ng) or norfloxacin (7–700 ng) on S. typhimurium TA102 were studied, employing the S9 mix (liver homogenate from rats pre-treated with Arochlor 1254) in the presence of 10–1000 μg of ascorbic acid (AA) with 0.1 mM FeCl3 or EDTA. Minimal inhibitory concentrations of NOR and NLX against 25 uropathogenic Escherichia coli strains were obtained using the plate dilution method in the presence of vitamin C. Vitamin C (1 mg) together with 0.1 mM FeCl3 showed a prooxidant effect in the S9 mix and enhanced the lipoperoxidation induced by either NOR or NLX. Mutagenic potency was also increased for both NOR and NLX. When metal ions were chelated with EDTA, ascorbate showed both antimutagenic and antioxidant properties. Mutagenic potency and lipoperoxidation were reduced for both NOR and NLX. The addition of vitamin C did not change the minimal in vitro inhibitory concentrations of NLX or NOR against the 25 uropathogenic E. coli strains. The antimutagenic and antioxidant effects of vitamin C were especially marked when the Salmonella strain was exposed to NOR or NLX in the presence of EDTA. In contrast, the vitamin C in the presence of FeCl3 increased ROS generation, enhancing both the mutagenic effect of the quinolones and malondialdehyde production from lipoperoxidation induced in the bacterial membranes. Therapeutic use of quinolones together with vitamin C and divalent cations might induce the Fenton reaction involving norfloxacin and nalidixic acid. However, our results suggest that vitamin C could be a good alternative for reducing the genotoxic risk of these therapeutic drugs if it is carefully handled.Kinoloni su antibiotici širokog spektra koji djeluju na Gram-pozitivne i Gram-negativne bakterije. Stvaraju reaktivne metabolite kisika što mogu oštetiti strukturu stanica i time narušiti zdravlje ljudi. Da bi se taj rizik smanjio, koristi se vitamin C koji međutim može imati i suprotno djelovanje: u prisutnosti dvovalentnih iona metala potiče Fentonovu reakciju, što dovodi do stvaranja hidroksil radikala (HO˙) koji oštećuju stanicu. Svrha je ovoga rada utvrditi antioksidativna i prooksidativna svojstva vitamina C ispitivanjem njegova utjecaja na peroksidaciju lipida i mutagenezu induciranu kinolonima poput nalidiksinske kiseline i norfloksacina iz soja bakterije Salmonella typhimurium TA102. Mutagenost je procijenjena Amesovim testom, a rezultati su izraženi kao broj revertanata histidin+/ng kinolona. Peroksidacija lipida mjerena je reakcijom produkata s tiobarbiturnom kiselinom, te izražena kao μmol malonaldehida/(mL·h). Istražen je utjecaj različitih koncentracija nalidiksinske kiseline (10-1000 ng) ili norfloksacina (7-700 ng) na bakteriju S. typhimurium TA102 pomoću mješavine S9 (homogenat jetre štakora tretiranih aroklorom 1254), 10-1000 μg vitamina C i 0,1 mM FeCl3 ili etilendiamintetraoctene kiseline (EDTA). Najmanje koncentracije norfloksacina i nalidiksinske kiseline, potrebne za inhibiciju 25 uropatogenih sojeva bakterije Escherichia coli, određene su metodom na agaru u prisutnosti vitamina C. Vitamin C (1 mg) uz 0,1 mM FeCl3 imao je prooksidativno djelovanje u mješavini S9, pospješio je peroksidaciju lipida uzrokovanu norfloksacinom ili nalidiksinskom kiselinom, te pojačao mutagenost oba kinolona. Stvaranje kelatnog kompleksa iona metala s EDTA potaknulo je antimutageno i antioksidativno djelovanje vitamina C, te smanjilo mutageno i lipoperoksidacijsko djelovanje norfloksacina i nalidiksinske kiseline. Dodatak vitamina C nije smanjio minimalnu koncentraciju nalidiksinske kiseline i norfloksacina in vitro, potrebnu za inhibiciju 25 uropatogenih sojeva bakterije E. coli. Antimutageno i antioksidativno djelovanje vitamina C bilo je osobito izraženo kada je soj bakterije Salmonella izložen djelovanju norfloksacina ili nalidiksinske kiseline u prisutnosti EDTA. Zajedno s FeCl3 vitamin C je ubrzao stvaranje reaktivnih metabolita kisika, pojačavajući time mutageno djelovanje kinolona te proizvodnju malondialdehida peroksidacijom lipida u membrani bakterija. Uporaba kinolona u kombinaciji s vitaminom C i dvovalentnim kationima pri liječenju bolesti mogla bi pokrenuti Fentonovu reakciju s norfloksacinom i nalidiksinskom kiselinom. Međutim, rezultati autora pokazuju da bi se vitamin C, pravilnom primjenom, mogao upotrijebiti kako bi se smanjila opasnost od genotoksičnog djelovanja tih lijekova

Prospective study of urinary tract infection surveillance after kidney transplantation

<p>Abstract</p> <p>Background</p> <p>Urinary tract infection (UTI) remains one of the main complications after kidney transplantation and it has serious consequences.</p> <p>Methods</p> <p>Fifty-two patients with kidney transplantation were evaluated for UTI at 3-145 days (mean 40.0 days) after surgery.. Forty-two received a graft from a live donor and 10 from a deceased donor. There were 22 female and 30 male patients, aged 11-47 years. Microscopic examinations, leukocyte esterase stick, and urinary culture were performed every third day and weekly after hospitalization. A positive culture was consider when patients presented bacterial counts up to 10⁵ counts.</p> <p>Results</p> <p>UTI developed in 19/52 (37%) patients at 3-75 days (mean 19.5 days after transplantation. Recurrent infection was observed in 7/52 (13.4%) patients at days 17-65. UTI was more frequent in patients who received deceased grafts compared with live grafts (7/10, 70% <it>vs</it>. 12/42, 28%; p < 0.007). Female patients were more susceptible than male (11/22, 50% <it>vs</it>. 8/22, 36.35%; p < 0.042). Five-year survival rate was 94.5% (49/52 patients). Kidney Graft exit update is 47/52 (90.2%), and there were no significant differences between graft rejection and UTI (p = 0.2518). Isolated bacteria were <it>Escherichia coli </it>(31.5%), <it>Candida albicans </it>(21.0%) and <it>Enterococcus </it>spp. (10.5%), followed by <it>Pseudomonas aeruginosa, Klebsiella pneumoniae, Morganella </it><it>morganii, Enterobacter cloacae </it>and <it>Micrococcus </it>spp. Secondary infections were produced by (7/19, 36.8%). <it>Enterococcus </it>spp. (57%), <it>E. coli </it>(28%) and <it>Micrococcus </it>spp. (14.2%). Antibiotic resistance was 22% for ciprofloxacin and 33% for ampicillin. Therapeutic alternatives were aztreonam, trimethoprim-sulfamethoxazole, netilmicin and fosfomycin.</p> <p>Conclusions</p> <p>Surveillance of UTI for the first 3 months is a good option for improving quality of life of kidney transplantation patients and the exit of graft function especially for female patients and those receiving deceased grafts. Antibiograms provided a good therapeutic alternative to patients who presented with UTIs after receiving a kidney allograft.</p

Higher COVID-19 pneumonia risk associated with anti-IFN-α than with anti-IFN-ω auto-Abs in children

We found that 19 (10.4%) of 183 unvaccinated children hospitalized for COVID-19 pneumonia had autoantibodies (auto-Abs) neutralizing type I IFNs (IFN-alpha 2 in 10 patients: IFN-alpha 2 only in three, IFN-alpha 2 plus IFN-omega in five, and IFN-alpha 2, IFN-omega plus IFN-beta in two; IFN-omega only in nine patients). Seven children (3.8%) had Abs neutralizing at least 10 ng/ml of one IFN, whereas the other 12 (6.6%) had Abs neutralizing only 100 pg/ml. The auto-Abs neutralized both unglycosylated and glycosylated IFNs. We also detected auto-Abs neutralizing 100 pg/ml IFN-alpha 2 in 4 of 2,267 uninfected children (0.2%) and auto-Abs neutralizing IFN-omega in 45 children (2%). The odds ratios (ORs) for life-threatening COVID-19 pneumonia were, therefore, higher for auto-Abs neutralizing IFN-alpha 2 only (OR [95% CI] = 67.6 [5.7-9,196.6]) than for auto-Abs neutralizing IFN-. only (OR [95% CI] = 2.6 [1.2-5.3]). ORs were also higher for auto-Abs neutralizing high concentrations (OR [95% CI] = 12.9 [4.6-35.9]) than for those neutralizing low concentrations (OR [95% CI] = 5.5 [3.1-9.6]) of IFN-omega and/or IFN-alpha 2

Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p &lt; 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Antimutagenic Effects of Vitamin C Against Oxidative Changes Induced by Quinolones

Quinolones are broad-spectrum antibiotics effective against both Gram-positive and Gram-negative bacteria. Reactive oxygen species (ROS) generated by quinolones may damage cell structures and could be a risk to health. The use of vitamin C to reduce such risks may have the opposite effects: vitamin C in the presence of divalent metal ions can induce the Fenton reaction, leading to hydroxyl radical (HO˙) generation and oxidative damage. The purpose of this study is to evaluate the antioxidant and prooxidant properties of vitamin C by measuring its effects on both lipid peroxidation and mutagenesis induced by quinolones nalidixic acid (NLX) or norfloxacin (NOR) in Salmonella typhimurium TA102. Mutagenicity was evaluated by the Ames test and the results were expressed as (histidine+ revertants/ng of quinolone), while lipoperoxidation was measured as thiobarbituric acid reactive substances (μmol malondialdehyde/(mL·h)). The effects of different concentrations of nalidixic acid (10–1000 ng) or norfloxacin (7–700 ng) on S. typhimurium TA102 were studied, employing the S9 mix (liver homogenate from rats pre-treated with Arochlor 1254) in the presence of 10–1000 μg of ascorbic acid (AA) with 0.1 mM FeCl3 or EDTA. Minimal inhibitory concentrations of NOR and NLX against 25 uropathogenic Escherichia coli strains were obtained using the plate dilution method in the presence of vitamin C. Vitamin C (1 mg) together with 0.1 mM FeCl3 showed a prooxidant effect in the S9 mix and enhanced the lipoperoxidation induced by either NOR or NLX. Mutagenic potency was also increased for both NOR and NLX. When metal ions were chelated with EDTA, ascorbate showed both antimutagenic and antioxidant properties. Mutagenic potency and lipoperoxidation were reduced for both NOR and NLX. The addition of vitamin C did not change the minimal in vitro inhibitory concentrations of NLX or NOR against the 25 uropathogenic E. coli strains. The antimutagenic and antioxidant effects of vitamin C were especially marked when the Salmonella strain was exposed to NOR or NLX in the presence of EDTA. In contrast, the vitamin C in the presence of FeCl3 increased ROS generation, enhancing both the mutagenic effect of the quinolones and malondialdehyde production from lipoperoxidation induced in the bacterial membranes. Therapeutic use of quinolones together with vitamin C and divalent cations might induce the Fenton reaction involving norfloxacin and nalidixic acid. However, our results suggest that vitamin C could be a good alternative for reducing the genotoxic risk of these therapeutic drugs if it is carefully handled