Quinacrine,an Antimalarial Drug with Strong Activity Inhibiting SARS-CoV-2 Viral Replication In Vitro

Quinacrine (Qx), a molecule used as an antimalarial, has shown anticancer, antiprion, and antiviral activity. The most relevant antiviral activities of Qx are related to its ability to raise pH in acidic organelles, diminishing viral enzymatic activity for viral cell entry, and its ability to bind to viral DNA and RNA. Moreover, Qx has been used as an immunomodulator in cutaneous lupus erythematosus and various rheumatological diseases, by inhibiting phospholipase A2 modulating the Th1/Th2 response. The aim of this study was to evaluate the potential antiviral effect of Qx against denominated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Vero E6 cells. The cytotoxicity of Qx in Vero E6 cells was determined by the MTT assay. Afterwards, Vero E6 cells were infected with SARS-CoV-2 at different multiplicities of infections (MOIs) of 0.1 and 0.01 in the presence of Qx (0–30 µM) to determinate the half maximal effective concentration (EC₅₀). After 48 h, the effect of Qx against SARS-CoV-2 was assessed by viral cytotoxicity and viral copy numbers, the last were determined by digital real-time RT-PCR (ddRT-PCR). Additionally, electron and confocal microscopy of Vero E6 cells infected and treated with Qx was studied. Our data show that Qx reduces SARS-CoV-2 virus replication and virus cytotoxicity, apparently by inhibition of viral ensemble, as observed by ultrastructural images, suggesting that Qx could be a potential drug for further clinical studies against coronavirus disease 2019 (COVID-19) infection.     

Epidemiology of urinary tract infections: transmission and risk factors,incidence, and costs

Uropathogenic E. coli have special features that allow them to take advantage of the bladder environment. There UPEC can grow to substantial numbers in pure culture that are shed frequently into the environment and have a high probability of transmission to other hosts. A better understanding of the transmission system and the host and bacterial factors influencing transmission is essential for the identification of effective prevention strategies. Although the risk of severe morbidity among otherwise healthy populations is low, the incidence is high. Further, the potential impact of antibiotic treatment on the emergence of increasingly antibiotic-resistant UPEC (the resistant mechanisms that may be transmitted to other bowel inhabitants) is high. The benefits of successfully preventing UTI are substantial.     

In vivo evaluation of the dopaminergic neurotransmission system using [123I]FP‐CIT SPECT in 6‐OHDA lesioned rats

The 6‐hydroxydopamine (6‐OHDA) rodent model of Parkinson's disease (PD) has been used to evaluate the nigrostriatal pathway. The aim of this work was to explore the relationship between the degree of 6‐OHDA‐induced dopaminergic degeneration and [¹²³I]FP‐CIT binding using single photon emission computed tomography (SPECT). Fourteen rats received a 6‐OHDA injection (4 or 8 µg) into the left medial forebrain bundle. After 3 weeks, magnetic resonance imaging and scans with a small‐animal SPECT system were performed. Finally, the nigrostriatal lesion was assessed by immunohistochemical analysis. Immunohistochemical analysis confirmed two levels of dopaminergic degeneration. Lesions induced by 6‐OHDA diminished the ipsilateral [¹²³I]FP‐CIT binding by 61 and 76%, respectively. The decrease in tracer uptake between control and lesioned animals was statistically significant, as was the difference between the two 6‐OHDA lesioned groups. Results concluded that [¹²³I]FP‐CIT SPECT is a useful technique to discriminate the degree of dopaminergic degeneration in a rat model of PD. Copyright © 2014 John Wiley & Sons, Ltd.     

Distribution of APOE polymorphism in the “Paisa” population from northwest Colombia (Antioquia)

Background: The apolipoprotein E (APOE) gene plays a pivotal role in cholesterol metabolism. Since the discovery of the APOE*2 and APOE*4 as the major susceptibility alleles for several diseases including dyslipidemia, atherosclerosis, coronary heart disease, late-onset and early Alzheimer’s disease, the APOE genotype might be considered as a potential predictive factor for both epidemiological research and diagnosis.

Aim: The aim of this study is to report on the polymorphism of the APOE gene in the “Paisa” population from northwest Colombia (Antioquia) to obtain a population baseline of the existing variation in this locus.

Method: One thousand and one healthy voluntaries were genotyped for the APOE polymorphism using polymerase chain reaction–restriction fragment length polymorphism technique.

Results: The APOE*3/*3 genotype presented the highest frequency (66.33%) and the APOE*4/*4 had the lowest frequency (1.89%). Genotype frequencies comply with Hardy–Weinberg expectations. Allele frequencies obtained for APOE*2, APOE*3 and APOE*4 were 0.075?±?0.005 (95% CI?=?0.063–0.086), 0.814?±?0.009 (0.797–0.831) and 0.111?±?0.007 (0.098–0.125), respectively.

Conclusion: Although globally the high-to-low APOE frequency follows the E*3?>?E*4?>?E*2 trend, the present APOE frequency data is in disagreement with some reports from South-American countries.     

Dramatic Decrease in Prevalence of Soil-Transmitted Helminths and New Insights Into Intestinal Protozoa in Children Living in the Chaco Region,Bolivia

We assessed the prevalence of intestinal parasites among 268 2–12-year-old children living in rural areas, small villages, and semi-urban areas of the Chaco region, south-eastern Bolivia. The overall parasitism was 69%. Only protozoa, helminths, or co-infections were observed in 89.2%, 5.9%, or 4.9% of the positive children, respectively. A significant progressive increase in overall parasite prevalence was found when passing from rural areas to small villages and semi-urban areas. The most commonly found species were Entamoeba coli (38.4%), Giardia intestinalis (37.7%), and Blastocystis spp. (16%). Hymenolepis nana was the most prevalent helminth (5.6%), followed by Ascaris lumbricoides and hookworms (1.5% and 0.4%) evidenced only in rural areas and in villages. Molecular diagnostics identified Blastocystis subtypes 9 and 2, and 5 infections by Entamoeba histolytica and 4 by Entamoeba dispar. The dramatic decrease in prevalence of soil-transmitted helminths with respect to that observed about 20 years ago (> 40%) evidences the success of the preventive chemotherapy intervention implemented in 1986. Health education and improved sanitation should be intensified to control protozoan infections.In developing countries the lack of access to safe water, sanitation, and hygiene are the key factors for the high prevalence of intestinal protozoa that, in infants and children, frequently have the clinical expression of malabsorption syndrome and gastrointestinal morbidity.¹ Moreover, Ascaris lumbricoides may contribute to nutritional deficiencies and even produce intestinal occlusion, whereas other soil-transmitted helminths (STHs) cause chronic intestinal blood loss that results in anemia, and impairing physical growth, cognition, learning and working capacities.²In the Santa Cruz Department (Plurinational State of Bolivia), studies conducted ∼40 years ago showed intestinal parasitism ranging from 85.4% to 99.5%, with 65% of polyparasitism.^3–5 A survey carried out in 1987 detected an overall value of 79%, with prevalence of STHs up to 64%.^6,7 Further investigations conducted in 1990 in children living in two rural communities showed prevalence of STH infections of 41% and 64%, respectively.⁸ Starting in 1986, the Bolivian Ministry of Health developed a Parasitic Disease Control Program based on preventive chemotherapy with mebendazole that is still part of the Integral Attention to Prevalent Childhood Diseases Program (AIEPI).The study reported herein, programmed and carried out in 2011 with the support of the Guaraní political organization (Asamblea del Pueblo Guaraní) and in agreement with the Bolivian Ministry of Health (who gave the Ethical approval), was aimed at evaluating the current prevalence of intestinal parasites in apparently healthy children. Results of the analyses were reported daily, and positive subjects had immediate access to further specific medical check-up and drug treatment.A total of 268 randomly selected children (120 boys and 148 girls) 2–12 years of age were enrolled. Sample collection was performed in rural communities (Añimbo, Arenal, Brecha, Mandiyuti, Taputá, Timboirenda, Uruguay), small villages (San Antonio del Parapetí, Espino, Ivicuati), and semi-urban areas (Boyuibe, La Brecha, Cuevo, Lagunillas), ~3 months after the last delivering of preventive chemotherapy. Stool specimens daily collected, in the afternoon were submitted to microscopic examination, in a drop of iodine solution, of both wet smears and sediments after Ridley concentration.⁹ Parasites were identified on the basis of their morphological features.¹⁰ Samples positive for Entamoeba histolytica complex and for Blastocystis spp. were further analyzed by polymerase chain reaction (PCR) amplification and sequencing to identify species/subtypes involved. Genomic DNA was extracted using the NucleoSpin tissue kit (Macherey-Nagel, Düren, Germany) and a 540-bp (and in the nested PCR a 374-bp) fragment of the 30-kDa surface antigen from E. histolytica complex and the small subunit rRNA gene of Blastocystis spp. were PCR amplified following published protocols.^11,12 Amplicons were purified (SureClean kit, Aurogene, Rome, Italy) and sequenced (Eurofins MWG Operon, Ebersberg, Germany). Sequences obtained were corrected, aligned, and compared with Blastocystis SSU rRNA gene sequences available database (http://www.ncbi.nlm.nih.gov). Subtypes (STs) were identified according to the classification proposed by Stensvold and others in 2007.¹³Statistical analysis (chi-square [χ²] or Fisher''s exact tests) was applied to the results, and P values < 0.05 were considered significant.Intestinal protozoa and/or eggs from helminths were recovered in 185 of 268 children (69.0%). Only protozoa were evidenced in 165 subjects (89.2%). Helminths, and protozoa associated to helminths, were detected in 5.9% and 4.9% of the positive children, respectively. The most commonly found parasite was Entamoeba coli, followed by Giardia intestinalis, and Blastocystis spp. (