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Abstract Gene flow between cultivated and wild gene pools is common in the contact zone between agricultural lands and natural habitats and can be used to study the development of adaptations and selection of novel varieties. This is likely the case in the northern Adriatic region, where centuries-old cultivated orchards of sweet chestnut (Castanea sativa Mill.) are planted within the natural distribution area of the species. Thus, we investigated the population structure of several orchards of sweet chestnuts. Furthermore, the genetic background of three toponymous clonal varieties was explored. Six genomic simple sequence repeat (gSSR) and nine EST-derived SSR (EST-SSR) loci were utilized in this research, and both grafted and non-grafted individuals were included in this study. Five closely related clones were identified, which represent a singular, polyclonal marron variety, found in all three cultivation areas. Furthermore, many hybrids, a result of breeding between cultivated and wild chestnuts, have been found. Analyzed semi-wild orchards defined by a diverse genetic structure, represent a hotspot for further selection and could result in creation of locally adapted, high-yielding varieties.

IntroductionForest ecosystems are highly threatened by the simultaneous effects of climate change and invasive pathogens. Chestnut blight, caused by the invasive phytopathogenic fungus Cryphonectria parasitica, has caused severe damage to European chestnut groves and catastrophic dieback of American chestnut in North America. Within Europe, the impacts of the fungus are widely mitigated through biological control that utilizes the RNA mycovirus: Cryphonectria hypovirus 1 (CHV1). Viral infections, similarly to abiotic factors, can cause oxidative stress in their hosts leading to physiological attrition through stimulating ROS (reactive oxygen species) and NOx production.MethodsTo fully understand the interactions leading to the biocontrol of chestnut blight, it is vital to determine oxidative stress damage arising during CHV1 infection, especially considering that other abiotic factors, like long-term cultivation of model fungal strains, can also impact oxidative stress. Our study compared CHV1-infected C. parasitica isolates from two Croatian wild populations with CHV1-infected model strains (EP713, Euro7 and CR23) that have experienced long-term laboratory cultivation.Results and DiscussionWe determined the level of oxidative stress in the samples by measuring stress enzymes’ activity and oxidative stress biomarkers. Furthermore, for the wild populations, we studied the activity of fungal laccases, expression of the laccase gene lac1, and a possible effect of CHV1 intra-host diversity on the observed biochemical responses. Relative to the wild isolates, the long-term model strains had lower enzymatic activities of superoxide dismutase (SOD) and glutathione S-transferase (GST), and higher content of malondialdehyde (MDA) and total non-protein thiols. This indicated generally higher oxidative stress, likely arising from their decades-long history of subculturing and freeze–thaw cycles. When comparing the two wild populations, differences between them in stress resilience and levels of oxidative stress were also observed, as evident from the different MDA content. The intra-host genetic diversity of the CHV1 had no discernible effect on the stress levels of the virus-infected fungal cultures. Our research indicated that an important determinant modulating both lac1 expression and laccase enzyme activity is intrinsic to the fungus itself, possibly related to the vc type of the fungus, i.e., vegetative incompatibility genotype.

Blora Regency is a barren area in Indonesia and has drought disaster potential, to address those problems Indonesian Governments launched Randugunting Dam’s construction program. The research area was located at Block 37 – 40, Radugunting Dam, Japah District, Blora Region. Dam as a water retention building should have foundation rocks that are resistant towards seepage potential, therefore curtain grouting was conducted to seal geological fractures on the foundation rocks. The purpose of these research was to understand the seepage potential on foundation rocks, foundation rocks curtain grouting requirement, and curtain grouting effectiveness. Research method used were water pressure test (WPT) data analysis, in the form of water flow discharge data processed through calculation to produces Lugeon value (Lu). The Lugeon value (Lu) would be further analyzed to determine the curtain grouting effectiveness value. The result of the analysis shows that around 18.15% of the research area have seepage potential that was shown by medium (3 < Lu < 10) – high (Lu > 10) Lugeoun (Lu) value. After the curtain grouitng was conducted, it effectiveness value on Block 37 was 88.34% (good), on Block 38 was 81.43% (good), on Block 39 was 70,62% (good), and on Block 40 was 79.41% (good).

Cryphonectria parasitica is an invasive fungal pathogen that causes blight disease on chestnut trees. Its destructive effect can be controlled with naturally occurring mycovirus Cryphonectria hypovirus 1 (CHV1). To date, the spread of C. parasitica and CHV1 in Europe is fairly well documented, but there are still several unexplored regions. Thus, we sampled blight cankers from four sweet chestnut populations in Bay of Kotor and Lake Skadar regions in Montenegro. We determined vegetative compatibility (vc) type and mating-type diversity using molecular vic and MAT1 genotyping, as well as confirming the presence of CHV1 by RT-PCR. We identified 11 vc types, with EU-12 being the dominant one represented by 58.2% of all fungal isolates. The Shannon diversity index ranged from 0.93 to 1.47. Both mating types of C. parasitica were found in all four populations. The prevalence of CHV1 ranged from 15% to 40%. All CHV1 isolates belonged to Italian subtype I of CHV1 and were closely related, with relatively recent common ancestors. Our results indicate a longer presence of C. parasitica and CHV1 in Montenegro than previously thought. Natural biocontrol with CHV1 seems to be well established. However, it has the potential for deterioration; thus, close monitoring is required.

Laccase activity reduction in the chestnut blight fungus Cryphonectria parasitica usually accompanies the hypovirulence caused by the infection of fungus with Cryphonectria hypovirus 1 (CHV1). However, the different methods utilized for assessing this phenomenon has produced varied and often conflicting results. Furthermore, the majority of experimental setups included only one prototypic system, further confounding the results. Considering the diversity of fungal isolates, viral strains, and variability of their effects on the phytopathogenic process observed in nature, our goal was to ascertain if laccase activity variability is affected by (1) different C. parasitica isolates infected with several CHV1 strains, and (2) growth conditions. We have demonstrated that some CHV1 strains, contrary to previous assumptions, increase the activity of C. parasitica laccases. The specific fungal isolates used in the experiments and culture conditions also affected the results. Furthermore, we showed that two commonly used laccase substrates, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and 2,4-dimethoxyphenol, cannot be used interchangeably in C. parasitica laccase activity measurements. Our results illustrate the importance of conducting this type of study in experimental systems and culture conditions that resemble natural conditions as much as possible to be able to infer the most relevant conclusions applicable to natural populations.

This paper showcases the development of plant virology in Croatia at the University of Zagreb, Faculty of Science, from its beginning in the 1950s until today, more than 70 years later. The main achievements of the previous and current group members are highlighted according to various research topics and fields. Expectedly, some of those accomplishments remained within the field of plant virology, but others make part of a much-extended research spectrum exploring subviral pathogens, prokaryotic plant pathogens, fungi and their viruses, as well as their interactions within ecosystems. Thus, the legacy of plant virology in Croatia continues to contribute to the state of the art of microbiology far beyond virology. Research problems pertinent for directing the future research endeavors are also proposed in this review.

Ngempon Temple is one of the historical sites around the Mount Ungaran area. Ngempon Temple is the result of Hindu civilization in Indonesia which was built in the VIII-IX century. This research was conducted in the Ngempon area because it is necessary to conserve historical sites associated with geological sites in the study area. This study aims to determine the geological aspects that have the potential to become geotourism sites in the research area. The methods used are geological mapping for field data collection and petrographic analysis to determine rock characteristics. There are several sites including historical sites and geological sites in the vicinity of Ngempon Temple, namely Ngempon Temple, Diwak Waterfall, and Derekan Hot Springs. The lithologies found in the research area are claystone, tuff sandstone, basaltic andesite breccia, and andesite breccia. The volcanic rock in the research area is formed by Mount Ungaran activity during Lower Pleistocene until Holocene.

The research area was the exposed extruded igneous lava rock in Clering which became the mining location of PT Semarang Mineral Pembangunan in Clering, Donorejo, Jepara with an area of ± 11 ha. This study aims to identify and determine the composition of andesite minerals by macroscopic and microscopic, the main oxide compound data, the use of andesite in industrial sector. The lithology consisted of andesite lava textured, plagioclase, leucite, clinopyroxene, sanidine as phenocryst and ground mass in the form of microlithic andesite lava as a trachytic texture. The hardness of lithology tends to be more moderate, it is considered unsuitable for building foundations. Andesite and tuff in the study area are currently being mined to be used as raw material for glass because of high silica content. In the industrial sector, the feldspar minerals for flux glass, and ceramic raw materials with standard of PT Semarang Mineral Pembangunan and SNI ISO 14703: 2011, 1147-1984 and SNI ISO 12543: 2011. The lithology tuff can be utilized in the manufacturing of ceramics due to high silica and felspar content. The lithology andesite can be utilized as rocks flour in the manufacturing of fertilizer due to high natrium, potassium, and magnesium content.

Kayen District, Pati Regency is a karst morphology area composed of karstified limestone with geotourism potential, established as Karst Landscape Area. As basis for infrastructure development in geotourism area, a database of geosites need to be built to make sure it is available for government and public. The method used is field observation, interviews and SWOT analysis. Morphology of this area is divided into conical karst hills and the karst alluvium plains. The lithology of the research area are reef limestone, clastic limestone, and alluvial deposit. Inventory managed to identify potential geosites such as hills, cliffs, caves, and springs; five existing geosites have been developed and six locations are new geosite recommendations. From the SWOT analysis, it is known that this location has the strength that is the uniqueness of morphology from karstification results, the weakness is geotourism concept is still not yet understood by local residents, the opportunity is the minister's decision regarding the Sukolilo Karst Landscape Area and the threat is lack of geological protection at the research site. The geotourism and geotrack map will serve as geospatial information and further recommendation for government to prioritize the infrastructure development in this area.

The research area is a part of South Serayu Mountain zone, specifically Bregada Kulon Progo. The Bregada Kulon Progo is a product of volcanic activity in the past, which can be seen by the volcanic rock called Old Andesite Formation. The end of the volcanic activity is characterized by the deposition of carbonate rocks which were part of Jonggrangan Formation and Sentolo Formation. The lower part of Sentolo Formation is composed by the mix of carbonate and volcanic materials. The research is carried out by two methods. The first is field method, which is implemented by geological mapping and rock sampling. The second is laboratory method, which is implemented by petrographic method. The petrographic method is implemented by making thin section of the rocks, then observing the thin section with polarization microscope to determine the composition of the rocks. The rock samples are mainly from the lower and middle part of Sentolo Formation, which probably still got the influence of volcanic activity. The benefit of the research is knowing the influence of volcanic activity in the carbonate rocks of Sentolo Formation. The presence of volcanic materials in the lower part of Sentolo Formation, can be predicted by two processes. First, the volcanic activity supplied those materials directly, and second, from the rework of the volcanic materials then mixed with carbonat materials of Sentolo Formation.

This work analyzed exhaustion markers in CD8+ T-cell subpopulations in 21 samples of peripheral blood mononuclear cells (PBMCs) from individuals with ocular toxoplasmosis ( n = 9), chronic asymptomatic toxoplasmosis ( n = 7), and non-infected people ( n = 5) by using RT-qPCR and flow cytometry techniques. The study found that gene expression of PD-1 and CD244, but not LAG-3, was higher in individuals with ocular toxoplasmosis versus individuals with asymptomatic infection or uninfected. Expression of PD1 in CD8+ central memory (CM) cells was higher in nine individuals with toxoplasmosis versus five uninfected individuals ( p = .003). After ex vivo stimulation, an inverse correlation was found between the exhaustion markers and quantitative clinical characteristics (lesion size, recurrence index, and number of lesions). A total exhaustion phenotype was found in 55.5% (5/9) of individuals with ocular toxoplasmosis. Our results suggest that the CD8+ exhaustion phenotype is involved in the pathogenesis of ocular toxoplasmosis.

Background: Toxoplasmosis is potentially avoidable, treatable, and curable by simple and direct preventive measures. Knowledge, attitudes, and practices (KAP) assessments concerning gestational toxoplasmosis were evaluated in a cohort of pregnant women from Armenia-Quindío (Colombia, South America)., Methods: This cross-sectional descriptive KAP-type study was performed with informed consent between October 2021 and March 2022. The intervention involved a ten-minute talk administered by prenatal clinic nurses to pregnant women. This took place in the public health clinic RedSalud and the private clinic Happy Maternity with a post-KAP survey after pregnancy., Results: The findings of the initial KAP survey revealed that approximately 42.8 % of the 250 mothers surveyed had IgG anti-T. gondii antibodies present. A strong correlation was observed between a lower frequency of antibodies and a higher level of education. Following an educational intervention, 73 seronegative women demonstrated a significant improvement in their knowledge and behavior. Among the 111 mothers who received the intervention, 42 (37 %) were followed until delivery. Unfortunately, their level of compliance with prenatal serological follow-up was lower compared to previous historical records of cohort of mothers in the same health center during pre-pandemic periods. No seroconversion occurred, although the small number of cases makes the outcome inconclusive with respect to statistical significance., Conclusions: Education plays a crucial role in imparting valuable knowledge and fostering effective practices. It holds significant potential to prevent toxoplasmosis in pregnant seronegative mothers. Prenatal check-ups have proven to be a critical determinant in leveraging the benefits of education for seronegative mothers. Reporting and observed behaviors differed, identifying areas for improvement., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Women respond more favorably to biventricular pacing (BIVP) than men. Sex differences in atrioventricular and interventricular conduction have been described in BIVP studies. Left bundle branch area pacing (LBBAP) offers advantages due to direct capture of the conduction system. We hypothesized that men could respond better to LBBAP than BIVP., Objectives: This study aims to describe the sex differences in response to LBBAP vs BIVP as the initial cardiac resynchronization therapy (CRT)., Methods: In this multicenter prospective registry, we included patients with left ventricular ejection fraction ≤35% and left bundle branch block or a left ventricular ejection fraction ≤40% with an expected right ventricular pacing exceeding 40% undergoing initial CRT with LBBAP or BIVP. The composite primary outcome was heart failure-related hospitalization and all-cause mortality. The primary safety outcome included all procedure-related complications., Results: There was no significant difference in the primary outcome when comparing men and women receiving LBBAP (P = 0.46), whereas the primary outcome was less frequent in women in the BIVP group than men treated with BIVP (P = 0.03). The primary outcome occurred less frequently in men undergoing LBBAP (29.9%) compared to those treated with BIVP (46.5%) (P = 0.004). In women, the incidence of the primary endpoint was 24.14% in the LBBAP group and 36.2% in the BIVP group; however, this difference was not statistically significant (P = 0.23). Complication rates remained consistent across all groups., Conclusions: Men and women undergoing LBBAP for CRT had similar clinical outcomes. Men undergoing LBBAP showed a lower risk of heart failure-related hospitalizations and all-cause mortality compared to men undergoing BIVP, whereas there was no difference between LBBAP and BIVP in women., Competing Interests: Funding Support and Author Disclosures Dr Diaz has received speaker honoraria and has been a proctor for Medtronic for conduction system pacing. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose., (Copyright © 2024 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.)

Toxoplasmosis is the most prevalent parasitic zoonosis worldwide, causing ocular and neurological diseases. No vaccine has been approved for human use. We evaluated the response of peripheral blood mononuclear cells (PBMCs) to a novel construct of Toxoplasma gondii total antigen in maltodextrin nanoparticles (NP/TE) in individuals with varying infectious statuses (uninfected, chronic asymptomatic, or ocular toxoplasmosis). We analyzed the concentration of IFN-γ after NP/TE ex vivo stimulation using ELISA and the immunophenotypes of CD4 + and CD8 + cell populations using flow cytometry. In addition, serotyping of individuals with toxoplasmosis was performed by ELISA using GRA6-derived polypeptides. Low doses of NP/TE stimulation (0.9 μg NP/0.3 μg TE) achieved IFN-γ-specific production in previously exposed human PBMCs without significant differences in the infecting serotype. Increased IFN-γ expression in CD4 + effector memory cell subsets was found in patients with ocular toxoplasmosis with NP/TE but not with TE alone. This is the first study to show how T-cell subsets respond to ex vivo stimulation with a vaccine candidate for human toxoplasmosis, providing crucial insights for future clinical trials., (© 2024 García-López et al.)

Background: Intracardiac echocardiography (ICE) is increasingly used during left atrial appendage occlusion (LAAO) as an alternative to transesophageal echocardiography (TEE). The objective of this study is to evaluate the impact of ICE versus TEE guidance during LAAO on procedural characteristics and acute outcomes, as well the presence of peri-device leaks and residual septal defects during follow-up., Methods: All studies comparing ICE-guided versus TEE-guided LAAO were identified. The primary outcomes were procedural efficacy and occurrence of procedure-related complications. Secondary outcomes included lab efficiency (defined as a reduction in in-room time), procedural time, fluoroscopy time, and presence of peri-device leaks and residual interatrial septal defects (IASD) during follow-up., Results: Twelve studies (n = 5637) were included. There were no differences in procedural success (98.3% vs. 97.8%; OR 0.73, 95% CI 0.42-1.27, p = .27; I 2  = 0%) or adverse events (4.5% vs. 4.4%; OR 0.81 95% CI 0.56-1.16, p = .25; I 2  = 0%) between the ICE-guided and TEE-guided groups. ICE guidance reduced in in-room time (mean-weighted 28.6-min reduction in in-room time) without differences in procedural time or fluoroscopy time. There were no differences in peri-device leak (OR 0.93, 95% CI 0.68-1.27, p = 0.64); however, an increased prevalence of residual IASD was observed with ICE-guided versus TEE-guided LAAO (46.3% vs. 34.2%; OR 2.23, 95% CI 1.05-4.75, p = 0.04)., Conclusion: ICE guidance is associated with similar procedural efficacy and safety, but could result in improved lab efficiency (as established by a significant reduction in in-room time). No differences in the rate of periprocedural leaks were found. A higher prevalence of residual interatrial septal defects was observed with ICE guidance., (© 2023 Wiley Periodicals LLC.)

Background: Infection is the most dreaded complication of cardiac implantable electronic devices (CIEDs), particularly in patients undergoing high-risk procedures (eg, generator change, device upgrade, lead/pocket revision)., Objective: The purpose of this study was to describe the impact of chlorhexidine gluconate (CHG) pocket lavage in high-risk procedures., Methods: Patients from a prospective multicenter registry undergoing high-risk procedures were included. CHG lavage was performed by irrigating the generator pocket with 20 cc of 2% CHG without alcohol followed by and normal saline (NS) irrigation. Only NS irrigation was performed in the comparison group. The primary efficacy outcome was CIED-related infection at 12 months. The primary safety outcome was any CHG-associated adverse event. The secondary outcome was CIED infection during long-term follow-up. Propensity score matching (PSM) analysis was performed for the primary efficacy outcome., Results: A total of 1504 patients were included. At 12-month follow-up, the primary efficacy outcome occurred in 4 of 904 CHG (0.4%) and 14 of 600 NS (2.3%) subjects (log-rank P = .005). On multivariate analysis, the use of CHG irrigation was associated with a lower risk of infection at 1-year follow-up (Cox proportional hazard ratio [HR] 0.138; 95% confidence interval [CI] 0.04-0.45; P = .001). This effect persisted during long-term follow-up. PSM demonstrated a significant reduction in CIED-related infection for the CHG group (0.2% vs 2.5%; Cox proportional HR 0.08; 95% CI 0.01-0.59; P = .014). No adverse events were associated with the use of CHG., Conclusion: CHG lavage during high-risk procedures was associated with a reduction in CIED-related infections without any adverse events reported. The benefits of CHG lavage were observed even during long-term follow up and in PSM analysis., (Copyright © 2023 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Background: Tranexamic acid (TXA) is used to reduce bleeding in body contouring procedures; however, there are no studies that show the effectiveness of TXA when it is also used in the immediate postoperative period., Methods: A controlled, randomized, parallel, and open-label clinical trial was carried out in adult patients undergoing liposculpture and/or abdominoplasty. A control group administering presurgical TXA and a study group with presurgical and postsurgical TXA were formed. The decrease in hemoglobin and the incidence of blood transfusions between both groups were compared as well as the possible adverse effects of TXA., Results: Four hundred twenty-seven subjects were included, 208 (48.7%) in the control group and 219 (51.3%) in the study group. The median age was 34 years (interquartile range 28-42). Median postoperative hemoglobin levels at 24 hours were similar in both groups (study 11.3 g/dL versus control 11.1 g/dL, P = 0.07); however, at 72 hours, postoperative hemoglobin was higher in the study group versus control (10.8 versus 10.0 g/dL, P ≤ 0.001). The incidence of transfusions at 72 hours was 1.8% in the study group and 8.6% in the control group, for a risk ratio of 0.21 (95% confidence interval 0.07-0.61). There were no adverse or thromboembolic events., Conclusion: TXA proved to be more effective in reducing intra- and postsurgical bleeding and the need for transfusions, when used preoperatively and continued for 48 hours after surgery, than when used only preoperatively, without reporting adverse or thromboembolic effects., Competing Interests: The authors have no financial interest to declare in relation to the content of this article., (Copyright © 2023 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of The American Society of Plastic Surgeons.)

Ocular toxoplasmosis (OT) is characterized by inflammation within the eye and is the most recognized clinical manifestation of toxoplasmosis. The objective of this study was to identify new single-nucleotide polymorphisms (SNPs) in the P2RX7 gene that may have significance in the immune response to OT in Colombian patients. A case-control study was conducted to investigate the associations between SNPs (rs1718119 and rs2230912) in the P2RX7 gene and OT in 64 Colombian patients with OT and 64 controls. Capillary electrophoresis was used to analyze the amplification products, and in silico algorithms were employed to predict deleterious SNPs. Stability analysis of amino acid changes indicated that both mutations could lead to decreased protein structure stability. A nonsynonymous SNP, Gln460Arg, located in the long cytoplasmic tail of the receptor, showed a significant association with OT (Bonferroni correction (BONF) = 0.029; odds ratio OR = 3.46; confidence interval CI: 1.05 to 11.39), while no significant association between rs1718119 and OT risk was observed. Based on the 3D structure analysis of the P2RX7 protein trimer, it is hypothesized that an increase in the flexibility of the cytoplasmic domain of this receptor could alter its function. This SNP could potentially serve as a biomarker for identifying Colombian patients at risk of OT.

Background: Left bundle branch area pacing (LBBAP) for cardiac resynchronization therapy (CRT) is an alternative to biventricular pacing (BiVp)., Objectives: The purpose of this study was to compare the outcomes between LBBAP and BiVp as an initial implant strategy for CRT., Methods: In this prospective multicenter, observational, nonrandomized study, first-time CRT implant recipients with LBBAP or BiVp were included. The primary efficacy outcome was a composite of heart failure (HF)-related hospitalization and all-cause mortality. The primary safety outcomes were acute and long-term complications. Secondary outcomes included postprocedural New York Heart Association functional class and electrocardiographic and echocardiographic parameters., Results: A total of 371 patients (median follow-up of 340 days [IQR: 206-477 days]) were included. The primary efficacy outcome occurred in 24.2% in the LBBAP vs 42.4% in the BiVp (HR: 0.621 [95% CI: 0.415-0.93]; P = 0.021) group, driven by a reduction in HF-related hospitalizations (22.6% vs 39.5%; HR: 0.607 [95% CI: 0.397-0.927]; P = 0.021) without significant difference in all-cause mortality (5.5% vs 11.9%; P = 0.19) or differences in long-term complications (LBBAP: 9.4% vs BiVp: 15.2%; P = 0.146). LBBAP resulted in shorter procedural (95 minutes [IQR: 65-120 minutes] vs 129 minutes [IQR: 103-162 minutes]; P < 0.001) and fluoroscopy times (12 minutes [IQR: 7.4-21.1 minutes] vs 21.7 minutes [IQR: 14.3-30 minutes]; P < 0.001), shorter QRS duration (123.7 ± 18 milliseconds vs 149.3 ± 29.1 milliseconds; P < 0.001), and higher postprocedural left ventricular ejection fraction (34.1% ± 12.5% vs 31.4% ± 10.8%; P = 0.041)., Conclusions: LBBAP as an initial CRT strategy resulted in a lower risk of HF-related hospitalizations compared to BiVp. A reduction in procedural and fluoroscopy times, shorter paced QRS duration, and improvements in left ventricular ejection fraction compared with BiVp were observed., Competing Interests: Funding Support and Author Disclosures Dr Diaz has received speaker honoraria and is a proctor for Medtronic for conduction system pacing. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose., (Copyright © 2023 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.)

Background: Epicardial access (EA) has emerged as an increasingly important approach for the treatment of ventricular arrhythmias and to perform other interventional cardiology procedures. EA is frequently underutilized because the current approach is challenging and carries a high risk of life-threatening complications., Objective: The purpose of this study was to determine the efficacy and safety of the SAFER (Sustained Apnea for Epicardial Access With Right Ventriculography) epicardial approach., Methods: Consecutive patients who underwent EA with the SAFER technique were included in this multicenter study. The primary efficacy outcome was the successful achievement of EA. The primary safety outcomes included right ventricular (RV) perforation, major hemorrhagic pericardial effusion (HPE), and bleeding requiring surgical intervention. Secondary outcomes included procedural characteristics and any complications. Our results were compared with those from previous studies describing other EA techniques to assess differences in outcomes., Results: A total of 105 patients undergoing EA with the SAFER approach from June 2021 to February 2023 were included. EA was used for ventricular tachycardia ablation in 98 patients (93.4%), left atrial appendage closure in 6 patients (5.7%), and phrenic nerve displacement in 1 patient (0.9%). EA was successful in all subjects (100%). The median time to EA was 7 minutes (IQR: 5-14 minutes). No cases of RV perforation, HPE, or need of surgical intervention were observed in this cohort. Comparing our results with previous studies about EA, the SAFER epicardial approach resulted in a significant reduction in major pericardial bleeding., Conclusions: The SAFER epicardial approach is a simple, efficient, effective, and low-cost technique easily reproducible across multiple centers. It is associated with lower complication rates than previously reported techniques for EA., Competing Interests: Funding Support and Author Disclosures All authors have reported that they have no relationships relevant to the contents of this paper to disclose., (Copyright © 2023 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.)

Toxoplasma gondii (T. gondii) is a zoonotic apicomplexan parasite that is an important cause of clinical disability in humans. On a global scale, one third of the human population is infected with T. gondii. Mice and other small rodents are believed to be responsible for transmission of T. gondii to the domestic cat, its definitive host. Interferon-inducible Immunity-Related GTPases (IRG proteins) are important for control of murine T. gondii infections. Virulence differences between T. gondii strains are linked to polymorphic rhoptry proteins (ROPs) that cooperate to inactivate individual IRG family members. In particular, the pseudokinase ROP5 isoform B is critically important in laboratory strains of mice. We identified T. gondii ROP39 in complex with ROP5B and demonstrate its contribution to acute T. gondii virulence. ROP39 directly targets Irgb10 and inhibits homodimer formation of the GTPase leading to an overall reduction of IRG protein loading onto the parasitophorous vacuolar membrane (PVM). Maintenance of PVM integrity rescues the parasite from IRG protein-mediated clearance in vitro and in vivo. This study identifies a novel T. gondii effector that is important for specific inactivation of the IRG resistance system. Our data reveal that yet unknown T. gondii effectors can emerge from identification of direct interaction partners of ROP5B., Competing Interests: The authors declare that they have no conflict of interest., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Two zoonotic protozoan pathogens, Giardia duodenalis and Toxoplasma gondii , are important causes of waterborne infections in the Quindío region in Colombia. No previous data exist on how contamination occurs at the source for drinking water consumed by the human population in this region. Our aim was to describe the frequency of G. duodenalis and T. gondii DNA in 11 sampling points during a five-month period in water and adjacent soil at the Quindío River basin (Andean region in the central western part of Colombia). The study employed nested PCR for T. gondii , using the B1 gene as the amplification target, and single-round PCR for G. duodenalis assemblage A and assemblage B, amplifying the gdh gene, followed by DNA sequencing. In 50 soil samples, 28% (14/50) were positive for T. gondii. For G. duodenalis , distribution was in equal parts for assemblage A (8%; 4/50) and assemblage B (8%, 4/50). Genotyping of T. gondii sequences showed two soil samples with type I strain, another two samples of soil with type III strain, but most samples were of unidentified strains. In water samples, T. gondii was detected in 9.1% (5/55), G. duodenalis assemblage A in 34.5% (19/55), and G. duodenalis assemblage B in 12.7% (7/55). T. gondii DNA positivity was associated with lower soil temperature ( p  = 0.0239). Presence of G. duodenalis and T. gondii was evidenced in soil and water samples in the Quindío River basin, indicating soil as the potential source of contamination for the river that it is destined for human consumption. Monitoring these protozoa in drinking water is necessary to prevent public health risks in human populations., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Toxoplasma gondii, as other apicomplexa, employs adhesins transmembrane proteins for binding and invasion to host cells. Search and characterization of adhesins is pivotal in understanding Apicomplexa invasion mechanisms and targeting new druggable candidates. This work developed a machine learning software called ApiPredictor UniQE V2.0, based on two approaches: support vector machines and multilayer perceptron, to predict adhesins proteins from amino acid sequences. By using ApiPredictor UniQE V2.0, five SAG-Related Sequences (SRSs) were identified within the Toxoplasma gondii proteome. One of those candidates, TgSRS12B, was cloned in plasmid pEXP5-CT/TOPO and expressed in E. coli BL21 DE3. The resulting recombinant protein was purified via affinity chromatography. Co-precipitation assays in CaCo and Muller cells showed interactions between TgSRS12B-His-tagged and the membrane fractions from both human cell lines. In conclusion, we demonstrated that ApiPredictor UniQE V2.0, a bioinformatic free software, was able to identify TgSRS12B as a new adhesin protein., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Aim: To determine the professional profile and the work conditions of nurses working in intensive care units (ICU) in Colombia, Argentina, Peru and Brazil., Background: ICUs require a differentiated professional profile to provide quality care, and appropriate working conditions, leading to a transformation of care and management practices., Design: Descriptive multicentre cross-sectional observational study., Methods: An online survey was applied to identify both the characteristics of the professional profile and the working conditions. 1,427 ICU nursing professionals were included. RStudio statistical software was used for the analysis of the information. Descriptive statistics were used for the presentation of the results. The STROBE checklist for cross-sectional studies was used in this study., Results: Only 33.6% of the professionals had a specialisation degree in intensive care. The skills that were most frequently put into practice were communication (68.5%) and care management (78.5%). The most predominant nurse-to-patient ratios were 1:2, and greater than 1:6. 59.1% of the nurses had an indefinite term contract, 38.8% worked 48 hours per week and 49.8% had rotating shifts. Only 50.4% of them received incentives. The average salary ranged between 348 and 1,500 USD. 64.5% of the participants were satisfied with their job., Conclusion: It is necessary to strengthen nurses' professional profile by promoting both postgraduate education and the development of troubleshooting and teamwork skills. It is necessary to standardise the nurse-to-patient ratio, improve wages and increase incentives to achieve greater job satisfaction., Relevance to Practice: The knowledge and the improvement of both the professional profile and the work conditions of nurses working in intensive care units will improve the quality of the care given to critical patients and, therefore, the quality of health outcomes., (© 2021 The Authors. Journal of Clinical Nursing published by John Wiley & Sons Ltd.)

Aims: To evaluate the biological activity of extracts from cultures of marine bacteria against Toxoplasma gondii and Mycobacterium tuberculosis., Methods and Results: Ethyl acetate extracts obtained from seven marine bacteria were tested against T. gondii GFP-RH and M. tuberculosis H37Rv. The cytotoxicity on HFF-1 cells was measured by a microplate resazurin fluorescent approach, and the haemolytic activity was determined photometrically. The extracts from Bacillus sp. (INV FIR35 and INV FIR48) affected the tachyzoite viability. The extracts from Bacillus, Pseudoalteromonas, Streptomyces and Micromonospora exhibited effects on infection and proliferation processes of parasite. Bacillus sp. INV FIR48 extract showed an minimum inhibitory concentration value of 50 µg ml -1 against M. tuberculosis H37Rv. All the extracts exhibited relatively low toxicity to HFF-1 cells and the primary culture of erythrocytes, except Bacillus sp. INV FIR35, which decreased cell viability under 20%. Liquid chromatography coupled to mass spectrometry analysis of the most active bacterial extract Bacillus sp. INV FIR48 showed the presence of peptide metabolites related to surfactin., Conclusions: The extract from culture of deep-sea Bacillus sp. INV FIR48 showed anti-T. gondii and anti-tuberculosis (TB) biological activity with low cytotoxicity. In addition, peptide metabolites were detected in the extract., Significance and Impact of the Study: Toxoplasmosis and TB are among the most prevalent diseases worldwide, and the current treatment drugs exhibit side effects. This study confirm that marine bacteria are on hand sources of anti-infective natural products., (© 2021 The Society for Applied Microbiology.)

Purpose: To stablish if Blastocystis subtypes influences gastrointestinal symptoms., Methods: Case-control study. We obtained sequencing for Blastocysts subtyping from 13 patients with gastrointestinal symptoms (diarrhea or abdominal pain) and 12 from individuals without symptoms., Results: 12 sequences were from Subtype 2 and one from Subtype 3 in symptomatic individuals and nine samples were from Subtype 1, one from Subtype 2, and two from Subtype 3 in asymptomatic individuals. The prevalence of subtype 2 in symptomatic individuals was vastly different compared to the frequency in asymptomatic individuals (84.6% vs. 16.6%; OR 27.5 95% CI 3.2-233; Fisher exact test p = 0.0010201335). After in vitro culture, 22 isolates were obtained. Significant differences were observed for the 12 isolates from Subtype 2 that get a smaller number of total cells with dominant growth of vacuolar forms, compared with Subtypes 1 and 3, after eight days of culture., Conclusion: Our results suggest that gastrointestinal symptoms in Colombian individuals with Blastocystis infection depend on the infecting subtype with peculiar phenotypic characteristics in in vitro culture., (© 2021. Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Objectives: To determine the frequency of retinochoroidal lesions by ocular toxoplasmosis and their relationships with risk factors, in residents of two districts with high exposure to Toxoplasma, in Armenia-Quindío, Colombia., Methods: Cross-sectional analyses of fundoscopy screening, serological tests, and questionnaires were performed to determine risk factors associated with ocular toxoplasmosis retinochoroidal lesions. Differences in proportions were analyzed using the chi-squared test., Results: Of 161 individuals examined, 17 (10.5%) exhibited retinochoroidal scars suggestive of old inactive Toxoplasma gondii infection. All 17 individuals were seropositive for T. gondii antibodies. Consumption of bottled water was protective against T. gondii infection among individuals in this study. There were no specific epidemiological risk factors associated with ocular toxoplasmosis retinochoroidal lesions., Conclusion: Ocular toxoplasmosis is an important cause of visual impairment in Armenia-Quindío, Colombia. The consumption of boiled or bottled water is a major preventive public health measure to reduce infection by T. gondii and the subsequent onset of OT., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Published by Elsevier Ltd.)

We report diagnosis, treatment and evolution of cases of ocular toxocariasis in specialized consultation in Quindío, Colombia. No cases were seen during the 2000-17 period, but five cases were confirmed from November 2017 to March 2019; two children resulted with definitive loss of vision on the affected eye. Studies in contacts found that 12 of 19 (63%) family members and 15 of 25 (60%) children <15 years of age living on the same street were positive for IgG Toxocara antibodies. Epidemiological studies are necessaries to establish the reasons for the increase in cases at this region., (© The Author(s) [2020]. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Introduction: Toxoplasma gondii infections have been reported for many warm-blooded animals around the world including chiropterans. However, in Colombia, the country that holds the highest taxonomic richness of this order of mammals in the Neotropics, up to date there are no reports of T. gondii in bats (Carollia brevicauda)., Purpose: The objective of the present study was to detect T. gondii DNA from internal bat organs from Quindío, Colombia., Results: We report the first detection of T. gondii DNA from internal bat organs in the department of Quindio, Central Andes of Colombia. Out of three silky short tail bat (Carollia brevicauda) specimens collected at the natural reserve "La Montaña del Ocaso", organs were recovered (lungs, liver, heart, kidneys, small and large intestine) and tested for T. gondii through PCR for B1 sequence, with 1/3 (33.3%) positive result for the presence of T. gondii DNA in bat kidney tissues., Conclusion: Taking into consideration the high diversity of bat species in Colombia, and the complexity of the ecological and functional relationships that these organisms establish in the ecosystems they inhabit, we discuss on the urgent need for more detailed research and surveys for Toxoplansma in bats and other mammalian wild species.

Toxoplasma gondii is a parasite that can invade any cell in the human body. Here, we implemented and described an ex vivo model with human peripheral blood mononuclear cells (PBMCs) without using culture supplements/antibiotics and without cryopreserved cells (EXMOWS) to study the interactions between T. gondii and human cells. To establish the EXMOWS, three independent tests were carried out. Firstly, blood samples from 5 individuals were included to assess the viability and adherence of PBMCs in plate culture. In a second trial, blood samples from three seropositive and two seronegative individuals for T. gondii were used to evaluate human PBMCs cells: parasites, multiplicity of infection (MOI) 1:1, 1:3 and 1:5 at different times post infection (1 h, 6 h and 24 h). The possible immunomodulatory effect of the infection for this EXMOWS were evaluated in a third trial where HFF cells were infected with T. gondii and co-cultured with PBMCs obtained from anti-Toxoplasma IgG positive and IgG negative individuals. One hour was enough time for T. gondii infection of human PBMCs and 2 h was the minimum incubation time to guarantee adherence before carrying out any infection assay. A minimum of 1:3 MOI was necessary to guarantee efficient infection in human PBMCs with T. gondii RH-GFP. All protocols, including PBMCs isolation and stimulation, should be conducted the same day. This EXMOWS can be adapted to study the early stages of interaction with other microorganisms of human interest, without need of using cryopreservation and supplements/antibiotics., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Toxoplasma gondii , one of the world's most common parasites, can infect all types of warm-blooded animals, including one-third of the world's human population. Most current routine diagnostic methods are costly, time-consuming, and labor-intensive. Although T. gondii can be directly observed under the microscope in tissue or spinal fluid samples, this form of identification is difficult and requires well-trained professionals. Nevertheless, the traditional identification of parasites under the microscope is still performed by a large number of laboratories. Novel, efficient, and reliable methods of T. gondii identification are therefore needed, particularly in developing countries. To this end, we developed a novel transfer learning-based microscopic image recognition method for T. gondii identification. This approach employs the fuzzy cycle generative adversarial network (FCGAN) with transfer learning utilizing knowledge gained by parasitologists that Toxoplasma is banana or crescent shaped. Our approach aims to build connections between microscopic and macroscopic associated objects by embedding the fuzzy C-means cluster algorithm into the cycle generative adversarial network (Cycle GAN). Our approach achieves 93.1% and 94.0% detection accuracy for ×400 and ×1,000 Toxoplasma microscopic images, respectively. We showed the high accuracy and effectiveness of our approach in newly collected unlabeled Toxoplasma microscopic images, compared to other currently available deep learning methods. This novel method for Toxoplasma microscopic image recognition will open a new window for developing cost-effective and scalable deep learning-based diagnostic solutions, potentially enabling broader clinical access in developing countries. IMPORTANCE Toxoplasma gondii , one of the world's most common parasites, can infect all types of warm-blooded animals, including one-third of the world's human population. Artificial intelligence (AI) could provide accurate and rapid diagnosis in fighting Toxoplasma So far, none of the previously reported deep learning methods have attempted to explore the advantages of transfer learning for Toxoplasma detection. The knowledge from parasitologists is that the Toxoplasma parasite is generally banana or crescent shaped. Based on this, we built connections between microscopic and macroscopic associated objects by embedding the fuzzy C-means cluster algorithm into the cycle generative adversarial network (Cycle GAN). Our approach achieves high accuracy and effectiveness in ×400 and ×1,000 Toxoplasma microscopic images., (Copyright © 2020 Li et al.)

Introduction: Cancer patient satisfaction with the healthcare team is of great relevance for assessing the quality of the care provided by the health system. In Mexico, no valid and reliable tool is available to assess this construct., Objective: To validate the Functional Assessment of Chronic Illness Therapy-Treatment Satisfaction-Patient Satisfaction (FACIT-TS-PS) instrument, version 4, in cancer patients., Method: Cross-sectional design, non-probability convenience sampling. The sample consisted of 200 cancer-diagnosed patients, with mean age of 45.86 ± 15.01 years. Exploratory and confirmatory factor analyses were conducted., Results: The exploratory factor analysis identified four factors, with a Cronbach alpha of 0.945, and an explained variance of 68.15 %. The confirmatory factor analysis indicated that the proposed theoretical model adjusts to the data with an error close to zero and, in addition, it is balanced and carefully measures overall patient satisfaction with the treatment., Conclusion: FACIT-TS-PS was shown to be a valid and reliable instrument for use in clinical care and research in Mexican cancer patients. Its use is recommended in the evaluation of oncology multidisciplinary healthcare teams in Mexico., (Copyright: © 2019 Permanyer.)

Toxoplasma gondii ROP16 and ROP18 proteins have been identified as important virulence factors for this parasite. Here, we describe the effect of ROP16 and ROP18 proteins on peripheral blood mononuclear cells (PBMCs) from individuals with different clinical status of infection. We evaluated IFN-γ, IL-10, and IL-1β levels in supernatants from PBMCs cultures infected with tachyzoites of the T. gondii wild-type RH strain or with knock-out mutants of the rop16 and rop18 encoding genes (RHΔ rop16 and RHΔ rop18 ). Cytokine secretion was compared between PBMCs obtained from seronegative individuals ( n = 10), with those with chronic asymptomatic ( n = 8), or ocular infection ( n = 12). We also evaluated if polymorphisms in the genes encoding for IFN -γ, IL-10, IL-1 β, Toll-like receptor 9 ( TLR9 ), and purinoreceptor P2RX7 influenced the production of the encoded proteins after ex vivo stimulation. In individuals with chronic asymptomatic infection, only a moderate effect on IL-10 levels was observed when PBMCs were infected with RHΔ rop16 , whereas a significant difference in the levels of inflammatory cytokines IFN-γ and IL-1β was observed in seronegative individuals, but this was also dependent on the host's cytokine gene polymorphisms. Infection with ROP16-deficient parasites had a significant effect on IFN-γ production in previously non-infected individuals, suggesting that ROP16 which is considered as a virulence factor plays a role during the primary infection in humans, but not in the secondary immune response., (Copyright © 2019 Hernández-de-los-Ríos, Murillo-Leon, Mantilla-Muriel, Arenas, Vargas-Montes, Cardona, de-la-Torre, Sepúlveda-Arias and Gómez-Marín.)

We assessed the risk for toxoplasmosis in 10 school restaurants in Armenia (Quindio, Colombia). We analyzed the presence of Toxoplasma gondii DNA in the food, water, and living and inert surfaces of school restaurants, and we correlated these findings with the results of food safety inspection scores and with the prevalence of specific anti-T. gondii antibodies in children who ate at these restaurants. Of the 213 samples, 6.1% were positive using PCR to test for T. gondii DNA. Positive samples were found in meat, water, cucumber, guava juice, inert surfaces, and living surfaces. In 60% (6/10) of the public school restaurants, there was at least one PCR T. gondii-positive sample. In 311 serum samples from children who attended the restaurants, 101 (33%) were positive for IgG and 12 (3.9%) for IgM anti-T. gondii. The median of the compound score for the fulfillment of inspection for food safety conditions was of 60.7% (range 50-72). Higher T. gondii PCR positivity in surfaces, food, or water at each restaurant was correlated with lower inspection scores for water supply and water storage conditions. Lower scores in physical infrastructure and disinfection procedures and higher scores in furniture were correlated with a higher prevalence of IgG anti-T. gondii in children who ate at those restaurants. Inspection scores can identify restaurants with a higher risk for the presence of T. gondii.

Toxoplasma gondii engenders the common parasitic disease toxoplasmosis in almost all warm-blooded animals. Being a critical secretory protein, ROP18 is a major virulence factor of Toxoplasma . There are no reports about ROP18 detection in human serum samples with different clinical manifestations. New aptamers against ROP18 protein were developed through Systematic Evolution of Ligands by Exponential enrichment (SELEX). An Enzyme-Linked Aptamer Assay (ELAA) platform was developed using SELEX-derived aptamers, namely AP001 and AP002. The ELAA was used to evaluate total antigen from T. gondii RH strain (RH Ag) and recombinant protein of ROP18 (rROP18). The results showed that the ELAA presented higher affinity and specificity to RH Ag and rROP18, compared to negative controls. Detection limit of rROP18 protein in serum samples was measured by standard addition method, achieving a lower concentration of 1.56 μg/mL. Moreover, 62 seropositive samples with different clinical manifestations of toxoplasmosis and 20 seronegative samples were tested. A significant association between ELAA test positive for human serum samples and severe congenital toxoplasmosis was found ( p = 0.006). Development and testing of aptamers-based assays opens a window for low-cost and rapid tests looking for biomarkers and improves our understanding about the role of ROP18 protein on the pathogenesis of human toxoplasmosis., (Copyright © 2019 Vargas-Montes, Cardona, Moncada, Molina, Zhang and Gómez-Marín.)

Methods to detect protozoa are needed for food safety monitoring. We evaluated protocols to recover Giardia spp. cysts in Brassica oleracea (cabbage) and Lactuca sativa (lettuce) and then detection was performed by concentrating with formalin/ether solutions and microscopy or immunofluorescence or DNA amplification via PCR. To evaluate this methodology, G. duodenalis cysts were inoculated in triplicate (10 cysts) in 35-g samples of lettuce and cabbage. The method obtaining the highest percentage of recovery in cabbage was sulfamic acid solution plus stirring with stomacher (47.7% ± 7.5). For lettuce, the best method was glycine solution plus stirring with stomacher (46.6% ± 5.3). Inter-observer agreement was of 0.99. Giardia was detected by amplifying specific sequences for the DNA coding SSU rRNA. In 27 lettuce samples and 27 cabbage samples, obtained from supermarkets and street vendors, two lettuce samples (7.4%) and one cabbage sample (3.7%) were positive for Giardia via PCR assay and were sequenced, determining that they were two of assemblage B and one of lettuce to assemblage E. This method is proposed to detect Giardia in vegetables by PCR detection, enabling public health authorities to identify genotypes circulating in food, which will help to establish measures that reduce outbreaks of parasitic diseases associated with contaminated food.

Introduction: Toxoplasma gondii infection manifests differently in humans according to their immunity ranging from asymptomatic profiles to severe disease. There are multiple transmission mechanisms including blood transfusions, but little is known about the frequency of T. gondii infection in Colombia’s blood banks. Objective: To determine the prevalence of T. gondii infection in blood donors of a blood bank in the city of Cúcuta by serological and molecular diagnostic techniques. Materials and methods: We identified IgG and IgM antibodies against T. gondii by immunoassay in serum from 348 donors. The frequency of T. gondii DNA was determined by polymerase chain reaction (PCR) in whole blood from seropositive donors and relevant variables were analyzed based on the information obtained from surveys during blood donor selection. Results: Out of the 348 enrolled donors, 134 (38.5%) showed IgG antibodies against T. gondii; two of them (0.6%) had both IgG and IgM, and in two of them (1.5%), parasite DNA was detected in blood samples. A bivariate analysis indicated an association between seropositivity to T. gondii and being over 26 years of age (p=0.020). Conclusions: The prevalence of T. gondii infection found in the blood donors of this study suggests a significant exposure to the infectious agent that becomes relevant when parasitemia is detected.

Direct contact with swine infected by Aujeszky's disease virus (ADV) represents a potential risk of transmission to carnivore species, in which the infection is normally fatal. We assessed exposure to ADV in hunting dogs in an area where the virus is highly endemic in wild boar populations. Two out of 466 (0.43%; 95% CI 0.00-1.02%) hunting dogs analyzed were positive by gE-bELISA, gB-bELISA and the virus neutralization test. The seroprevalence levels detected, as well as the absence of reports of clinical cases in the hunting dog groups tested, indicate limited contact of this species with ADV in the study area. Further studies are warranted to assess the pathogenicity of Aujeszky's disease virus strains infecting wild boar.

Background: Making a definite diagnosis of infectious uveitis is a challenging task because many other infectious, and non-infectious uveitis, may have similar non-specific symptoms and overlapping clinical appearances. Co-infections in immunocompetent patients are not frequently proved with traditional serologic-diagnostic tools., Methods: Descriptive transversal study, in a Uveitis Service of an Ophthalmology Reference Center, in Bogotá, Colombia, from July 2014 to February 2016. Aqueous humor (AH) and/or vitreous fluid, blood and serum samples were collected from consecutive patients suspected of having infectious uveitis. The diagnosis of ocular toxoplasmosis (OT) was confirmed by the Goldmann-Witmer coefficient (GWC) and by polymerase chain reaction (PCR). Differential diagnosis by PCR in AH was done for viral origin such as Cytomegalovirus (CMV), Herpes simplex virus type 1 (HSV1), Herpes simplex virus type 2 (HSV2), Varicella zoster virus (VZV), Epstein-Barr virus (EBV) and Mycobacterium tuberculosis., Results: In 66 Colombian patients with uveitis of presumed infectious origin: 22 (33.3%) were confirmed as OT, 16 (24.2%) as undetermined OT, five (7.5%) as co-infections and 23 (34.8%) as other uveitis. Toxoplasma coinfection with M. tuberculosis was identified in one case by PCR and in four cases with HSV by GWC. The initial clinical diagnosis changed, after laboratory examination, in 21 cases (31.8%)., Conclusions: Clinical diagnosis can be changed by laboratory examination in a significant proportion of cases of uveitis. Diagnosis of OT should combine the use of PCR and GWC to reach the maximum of confirmation of cases. The use of multiple laboratory methods is necessary to identify co-infections and viral infections that can mimic OT in immunocompetent patients.

Background: Thiazolidinone derivatives show inhibitory activity (IC 50 ) against the Toxoplasma gondii parasite, as well as high selectivity with high therapeutic index. To disclose the target proteins of the thiazolidinone core in this parasite, we explored in silico the active sites of different T. gondii proteins and estimated the binding-free energy of reported thiazolidinone molecules with inhibitory effect on invasion and replication of the parasite inside host cells. This enabled us to describe some of the most suitable structural characteristics to design a compound derived from the thiazolidinone core., Results: The best binding affinity was observed in the active site of kinase proteins, we selected the active site of the T. gondii ROP18 kinase, because it is an important factor for the virulence and survival of the parasite. We present the possible effect of a derivative of thiazolidinone core in the active site of T. gondii ROP18 and described some characteristics of substituent groups that could improve the affinity and specificity of compounds derived from the thiazolidinone core against T. gondii., Conclusions: The results of our study suggest that compounds derived from the thiazolidinone core have a preference for protein kinases of T. gondii, being promising compounds for the development of new drugs with potential anti-toxoplasmosis activity. Our findings highlight the importance of use computational studies for the understanding of the action mechanism of compounds with biological activity.

Globally, congenital toxoplasmosis remains a significant cause of morbidity and mortality, and outbreaks of infection with T. gondii represent a significant, emerging public health burden, especially in the developing world. This parasite is a threat to public health. Disease often is not recognized and is inadequately managed. Herein, we analyze the status of congenital toxoplasmosis in Morocco, Colombia, the United States, and France. We identify the unique challenges faced by each nation in the implementation of optimal approaches to congenital toxoplasmosis as a public health problem. We suggest that developed and developing countries use a multipronged approach, modeling their public health management protocols after those in France. We conclude that education, screening, appropriate treatment, and the development of novel modalities will be required to intervene successfully in caring for individuals with this infection. Gestational screening has been demonstrated to be cost-effective, morbidity-sparing, and life-saving. Recognition of the value and promise of public health interventions to prevent human suffering from this emerging infection will facilitate better patient and societal outcomes.

Introduction: Cases of toxoplasmosis present in South America tend to be more severe than that found in other continents. Here, we present our clinical experience of ocular and ganglionar toxoplasmosis in the use of PCR, and of the treatment to prevent ocular involvement., Methodology: Retrospective analysis of clinical charts of patients with ocular and lymphadenitic toxoplasmosis at the parasitology and tropical medicine consultation in the "Universidad del Quindio" in Colombia. In total, 91 records of cases with ocular toxoplasmosis and 17 with lymphadenitis that underwent PCR analysis for B1 repeated sequence in blood, were compared to the results of 104 people with chronic asymptomatic toxoplasmosis. In addition, 41 clinical records were included from patients with confirmed toxoplasmic lymphadenitis: 10 untreated, 6 that begun treatment after four months of symptoms, and 25 that were treated during the first four months of symptoms and had a follow-up during at least one year., Results: Patients with ocular toxoplasmosis or lymphadenitis had a higher probability of PCR positivity in peripheral blood than chronic asymptomatic people. There were no cases of retinochoroiditis in 25 patients with toxoplasmic lymphadenitis treated before 4 months of symptoms and followed during at least 12 months. In four out of ten untreated cases, new lesions of retinochoroiditis presented after the symptoms of lymphadenitis., Conclusions: Toxoplasmosisin South America exhibits different clinical behavior and this influences the laboratory results as well as the need for treatment in the case of lymphadenitis. Clinicians should be aware of the geographical origin of the infection in order to adopt different therapeutic and diagnostic approaches., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In the past decade, the spread of emerging zoonotic flaviviruses (genus Flavivirus, family Flaviviridae) has been reported in many regions worldwide, representing a threat to both human and animal health. A serosurvey was carried out to assess exposure and risk factors associated with antigenically related flaviviruses, particularly West Nile virus (WNV), Usutu virus (USUV) and tick-borne encephalitis virus (TBEV), in dogs in Spain. Flavivirus antibodies were detected in 39 of 815 dogs (4.8%; 95% CI: 3.3-6.3) by bELISA. Significantly higher seropositivity was observed in hunting dogs compared to pet dogs. Virus neutralization tests confirmed WNV-specific and TBEV-specific antibodies in 11 and 14 bELISA-positive dogs, respectively. This is the first serosurvey of WNV and TBEV in dogs in Spain and the first report of TBEV circulation in this country. The seropositivity obtained indicates widespread, but not homogeneous, distribution of WNV and TBEV in dogs in Spain. In 2013 and 2015, WNV-seropositive dogs were detected in those areas of Andalusia where the highest number of WNV outbreaks were reported in both horses and humans. Antibodies against TBEV have been found in dogs sampled in two different periods and regions in Spain. Serosurveillance in dogs could be a complementary way of monitoring the activity of emerging flaviviruses in Spain., (© 2018 Blackwell Verlag GmbH.)

Toxoplasmosis is caused by infection with the protozoan parasite Toxoplasma gondii , which has the capacity to infect all warm-blooded animals worldwide. Toxoplasmosis is a major cause of visual defects in the Colombian population; however, the association between genetic polymorphisms in cytokine genes and susceptibility to ocular toxoplasmosis has not been studied in this population. This work evaluates the associations between polymorphisms in genes coding for the cytokines tumor necrosis factor alpha (TNF-α) (rs1799964, rs1800629, rs1799724, rs1800630, and rs361525), interleukin 1β (IL-1β) (rs16944, rs1143634, and rs1143627), IL-1α (rs1800587), gamma interferon (IFN-γ) (rs2430561), and IL-10 (rs1800896 and rs1800871) and the presence of ocular toxoplasmosis (OT) in a sample of a Colombian population (61 patients with OT and 116 healthy controls). Genotyping was performed with the "dideoxynucleotide (ddNTP) primer extension" technique. Functional-effect predictions of single nucleotide polymorphisms (SNPs) were done by using FuncPred. A polymorphism in the IL-10 gene promoter (-1082G/A) was significantly more prevalent in OT patients than in controls ( P = 1.93e-08; odds ratio [OR] = 5.27e+03; 95% confidence interval [CI] = 3.18 to 8.739; Bonferroni correction [BONF] = 3.48e-07). In contrast, haplotype "AG" of the IL-10 gene promoter polymorphisms (rs1800896 and rs1800871) was present at a lower frequency in OT patients ( P = 7e-04; OR = 0.10; 95% CI = 0.03 to 0.35). The +874A/T polymorphism of IFN-γ was associated with OT ( P = 3.37e-05; OR = 4.2; 95% CI = 2.478 to 7.12; BONF = 6.07e-04). Haplotype "GAG" of the IL-1β gene promoter polymorphisms (rs1143634, rs1143627, and rs16944) appeared to be significantly associated with OT ( P = 0.0494). The IL-10, IFN-γ, and IL-1β polymorphisms influence the development of OT in the Colombian population., (Copyright © 2018 American Society for Microbiology.)