Omicron - The new SARS-CoV-2 challenge?

SARS-CoV-2 virus has infected nearly 300 M people worldwide and has been associated with over 6 M deaths by March 2022. Since the virus emergence in December 2019 in Wuhan, several new mutations have been described. The World Health Organization has developed a working name for these emerging variants according to their impact on the worldwide population. In this context a high alert has been paid to variants of concern (VOC) due to their high infectiousness and transmissibility patterns. The most recent VOC, Omicron (B.1.1.529), has become dominant in the shortest time ever and has placed Europe under an overwhelming and unprecedented number of new cases. This variant has numerous mutations in regions that are associated with higher transmissibility, stronger viral binding, affinity and antibody escape. Moreover, the mutations and deletions present in the spike protein suggest that the SARS-CoV-2 specific attachment inhibitors may not be the best option for Omicron therapy. Omicron is the dominant variant circulating worldwide and, at the end of February 2022, it was responsible for nearly all sequences reported to GISAID. Omicron is made up of several sublineages, where the most common are BA.1 and BA.2 (or Nextstrain clade 21K and 21L, respectively). At a global level, it is possible to say that the proportion of BA.2 has been increasing relative to BA.1 and in some countries it has been replacing it at high rates. In order to better assess the Omicron effectiveness on antibody escape, spread and infectious ability it is of the highest relevance to maintain a worldwide tight surveillance. Even though this variant has been associated with a lower death rate, it is important to highlight that the number of people becoming infected is concerning and that further unpredictable mutations may emerge as the number of infected people rises.

Innovation results of IAM planning in urban water services.

The requirement to provide urban water services continuously while infrastructures are ageing, imposes the need for increasingly sustainable infrastructure asset management (IAM). To achieve and maintain adequate levels of service, the AWARE-P IAM methodology has been applied in collaborative projects launched by the National Civil Engineering Laboratory, in partnership with IST (Technical University of Lisbon), Addition (software company) and several water utilities. The objective of these projects is to support urban water utilities in the development, implementation and maintenance of IAM plans. To guarantee the success of IAM planning, following the AWARE-P IAM methodology, utilities are required to: consider that the infrastructure has system behaviour and lifespan is indefinite and guarantee the full-alignment of IAM planning with organisation objectives. By analysing the strategic and tactical plans of participating utilities, the proposed methodology principles are discussed and supported. The main innovation results from the implementation of IAM planning are also presented and discussed, including the challenges of setting up an IAM process, together with the major benefits and drawbacks that come up when developing IAM plans. The results were demonstrated by the effective implementation of 16 strategic and 14 tactical IAM plans by the participating utilities.

Alternatives to classic solvents for the isolation of bioactive compounds from Chrysochromulina rotalis.

This paper demonstrates a sequential partitioning method for isolating bioactive compounds from Chrysochromulina rotalis using a polarity gradient, replacing classic and hazardous solvents with greener alternatives. Seventeen solvents were evaluated based on their Hansen solubility parameters and for having a similar polarity to the solvents they would replace, four of which were selected as substitutes in the classic fractionation process. Considering the fatty acid and carotenoid recovery yields obtained for each of the solvents, it has been proposed to replace hexane (HEX), toluene (TOL), dichloromethane (DCM) and n-butanol (BUT) with cyclohexane, chlorobenzene, isobutyl acetate and isoamyl alcohol, respectively. In addition, cytotoxic activity was observed when the TOL and DCM solvent extracts were tested against tumour cell lines, demonstrating the antiproliferative potential of compounds containing, for example, fucoxanthin, fatty acids, peptides, isoflavonoids or terpenes, among others.

Urban water infrastructure asset management - a structured approach in four water utilities.

Water services are a strategic sector of large social and economic relevance. It is therefore essential that they are managed rationally and efficiently. Advanced water supply and wastewater infrastructure asset management (IAM) is key in achieving adequate levels of service in the future, particularly with regard to reliable and high quality drinking water supply, prevention of urban flooding, efficient use of natural resources and prevention of pollution. This paper presents a methodology for supporting the development of urban water IAM, developed during the AWARE-P project as well as an appraisal of its implementation in four water utilities. Both water supply and wastewater systems were considered. Due to the different contexts and features of the utilities, the main concerns vary from case to case; some problems essentially are related to performance, others to risk. Cost is a common deciding factor. The paper describes the procedure applied, focusing on the diversity of drivers, constraints, benefits and outcomes. It also points out the main challenges and the results obtained through the implementation of a structured procedure for supporting urban water IAM.

Assessing the environmental performance of urban wastewater systems using the INSA model: application to the Algés-Alcântara wastewater system, in Portugal.

Although the application of complex integrated models to wastewater systems is useful, it is often difficult to implement and not always suitable for the design of new systems or for their rehabilitation. Integrated simple approaches that allow assessing the environmental performance of urban wastewater systems may be advantageous, especially during the initial phases of the system planning process. This paper presents an original, straightforward approach that can be used for planning, design and operation of urban wastewater systems. The INtegrated Simplified Approach (INSA) combines the concepts of performance indicators with mass balances and can be applied to wastewater systems as a management support tool, particularly in situations where there is lack of data, economic limitations or time constraints. The INSA was applied to the Algés-Alcântara wastewater system to evaluate its environmental performance and to simulate the individual or combined impact of the rehabilitation measures proposed, thus defining their priority. The results clearly indicate that, despite the investment already made upgrading the wastewater treatment plant (WWTP), the proposed interventions must be implemented to ensure an acceptable environmental performance of the system. In addition, the results demonstrate the significant pollution loads present in stormwater, frequently higher than the pollution loads discharged into receiving waters during dry weather.

Dietary fructose, fruits, fruit juices and glucose tolerance status in Japanese-Brazilians.

BACKGROUND AND AIMS: Evidence suggests that fructose and sweetened beverages may be a risk factor for obesity and type 2 diabetes, but the role of sweetened fruit juices in glucose disturbances has been minimally explored. The aim of this study was to examine the association of total fructose, fresh fruit and sweetened fruit juice intake with glucose tolerance homeostasis in Japanese-Brazilians. METHODS AND RESULTS: A total of 475 men and 579 women aged >or=30 years were evaluated in a cross-sectional population-based survey with a standardized protocol including a 2-h oral glucose tolerance test (WHO criteria). Habitual food consumption was obtained using a validated food frequency questionnaire for Japanese-Brazilians. After adjustments for potential confounding variables, the odds ratio (OR; 95%CI) for impaired glucose tolerance was 2.1 (1.0-4.5; P for trend=0.05) for the highest as compared to the lowest tertile intake of total fructose and 2.3 (1.1-5.1; P for trend=0.05) for the highest as compared to the lowest tertile intake of sweetened fruit juices. CONCLUSION: Our results showed that high intakes of dietary fructose and sweetened fruit juices, but not whole fresh fruits, were associated with impaired glucose tolerance among genetically susceptible individuals.

Dietary practices and nutritional status of 0-24-month-old children from Brazilian Amazonia.

OBJECTIVE: To assess the nutritional status and dietary practices of 0-24-month-old children living in Brazilian Amazonia. DESIGN: Cross-sectional study. Information on children's dietary intakes was obtained from diet history data. Weight and length were measured for anthropometric evaluation. Fe status was assessed using fasting venous blood samples; Hb, serum ferritin and soluble transferrin receptor concentrations were measured. SETTING: The towns of Assis Brasil and Acrelândia in the state of Acre, north-west Brazil. SUBJECTS: A total of sixty-nine randomly selected 0-24-month-old children. RESULTS: Of these children, 40.3 % were anaemic, 63.1 % were Fe-deficient, 28.1 % had Fe-deficiency anaemia and 11.6 % were stunted. Breast-feeding was initiated by 97.1 % of mothers, followed by early feeding with complementary foods. The dietary pattern reflected a high intake of carbohydrate-rich foods and cow's milk, with irregular intakes of fruit, vegetables and meat. All infants and 92.3 % of toddlers were at risk of inadequate Fe intakes. Fe from animal foods contributed on average 0.5 % and 14.3 % to total dietary Fe intake among infants and toddlers, respectively. CONCLUSIONS: Poor nutritional status and inadequate feeding practices in this study population reinforce the importance of exclusive breast-feeding during the first 6 months of life. Greater emphasis is required to improve the bioavailability of dietary Fe during complementary feeding practices.

Antifungal suscepitibility profile of candida spp. oral isolates obtained from denture wearers.

Denture stomatitis is an inflammatory condition that occurs in denture wearers and is frequently associated with Candida yeasts. Antifungal susceptibility profiles have been extensively evaluated for candidiasis patients or immunosupressed individuals, but not for healthy Candida carriers. In the present study, fluconazole, itraconazole, voriconazole, terbinafine and 5-flucytosin were tested against 109 oral Candida spp. isolates. All antifungal agents were effective against the samples tested except for terbinafine. This work might provide epidemiological information about Candida spp. drug susceptibility in oral healthy individuals.

Effect of folate, vitamin B6, and vitamin B12 intake and MTHFR C677T polymorphism on homocysteine concentrations of renal transplant recipients.

Plasma hyperhomocysteinemia (HHcy) is considered a risk factor for chronic allograft dysfunction (CAD), the main cause of functional loss in transplant recipients. Genetic polymorphisms that alter enzymes involved in homocysteine (Hcy) metabolism, such as methylenetetrahydrofolate reductase (MTHFR), and vitamin deficiency can result in HHcy. The objectives of this study were to investigate the relationship between HHcy and CAD development, and to evaluate the effect of intake of folate and vitamins B6 and B12 as well as MTHFR C677T polymorphism on Hcy concentrations. Ninety-eight renal transplant recipients including 48 showing CAD and 50 with normal renal function (NRF), were included in this cross-sectional study. Peripheral blood samples were collected for plasma Hcy quantification by liquid chromatography/sequential mass spectrometry (LC-MS/MS), and for MTHFR polymorphism analysis using polymerase chain reaction-restriction fragment length polymorphism. Dietary intake was evaluated using a nutritional questionnaire. HHcy (P=.002) and higher mean concentrations of Hcy (P=.029) were associated with CAD. An association was observed between HHcy and 677T variant allele in the CAD group (P=.0005). There was no correlation between Hcy concentration and folate, vitamin B6 or vitamin B12 intake in the CAD group. However, a negative correlation was observed between Hcy concentration and folate intake (P=.043), and also between Hcy concentration and vitamin B6 intake (P=.030) in the NRF group. According to our study, HHcy is associated with CAD development. In patients with CAD, MTHFR polymorphism seems to have a greater effect on the Hcy concentration than the vitamin intake. Increased folate and vitamin B6 intakes seem to reduce Hcy concentrations among transplant recipients with NRF, and could contribute to reducing the risk of CAD development.

Decellularized Amniotic Membrane Scaffold as a Pericardial Substitute: An In Vivo Study.

BACKGROUND: In the development of new biomaterials for pericardium substitute, acellular amniotic membrane (AAM) presents potential for new applications in regenerative medicine. We studied an AAM as a pericardial substitute to achieve a suitable, cost effective, abundant matrix for the purpose of using it as graft for tissue repair. METHODS: Twenty Wistar rats were randomly divided into 2 groups (n = 10/group) and had their pericardiums excised. In the experimental group, the excised pericardium segment was substituted by a 7-mm-diameter patch of decellularized AAM sutured to the lesion area. After 4 weeks, the heart's outer layer of both groups was evaluated. The structure and component characteristics of the scaffold were determined with the use of hematoxylin and eosi, Alizarin Red S, and immumohistochemical staining and scanning electron microscopy. RESULTS: Histopathologic examination of the AAM patches revealed that the integrity of the AAM was preserved, and no calcification was observed on the surface of the myocardium. We also observed thicker pericardium repair tissue in the AAM group compared with the control group. AAM patches, by virtue of their low immunogenicity, evoked minimal host-versus-graft reaction. CONCLUSIONS: We conclude that AAM appears to be an ideal substitute for pericardium lesions, because it is integrated into the biologic tissue owing to its low immunogenicity and its ability to diminish the occurrence of adhesions and scarring, increasing the pericardium thickness.

Peculiar sequence organization of kinetoplast DNA minicircles from Trypanosoma cruzi.

The sequences of two minicircles from the kinetoplast DNA of the CL strain and one of the Y strain of Trypanosoma cruzi are reported. These 1.4 kb molecules have a peculiar sequence organization, the most distinctive feature being the occurrence of a 120 bp sequence repeated four times, located at 0, 90, 180 and 270 degrees along each circle. We have termed these conserved regions in this species 'minirepeats'. Minirepeats have a 3-fold higher concentration of cytosine residues in comparison with the variable regions and contain the universal 12-mer motif GGGGTTGGTGTA present in all sequenced minicircles and which was shown to be involved in DNA replication. A consensus sequence of T. cruzi minirepeats was determined using the 20 minirepeats present in five known T. cruzi minicircle sequences. This consensus sequence contains regions which have been remarkably well preserved in strains which show great biological diversity. In addition a low level of intraminicircle sequence similarity was also observed within the variable region, but this similarity did not extend between strains. The abundance of conserved minirepeat sequences containing invariant restriction sites in T. cruzi cells may prove valuable for the development of new direct diagnostic methods for Chagas' disease based on DNA probe technology.

Nitric oxide-induced apoptotic cell death in the acute phase of Trypanosoma cruzi infection in mice.

Production of gamma-interferon (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in Trypanosoma cruzi-infected mice results in the activation of inducible nitric oxide synthase (iNOS) and in elevated nitric oxide (NO) synthesis, which is important for the macrophage trypanocidal activity. However, NO has been shown to be involved in suppression of host immunity. In the present investigation, we studied the role of NO in inducing apoptosis in cells from BALB/c mice acutely infected by T. cruzi. Splenocytes from infected mice had a reduced cell viability and elevated levels of spontaneous apoptosis after 48 h in culture. Inhibition of NO production by the addition of the L-arginine analog NG-monomethyl-L-arginine (L-NMMA), or of monoclonal antibodies (mAbs) to IFN-gamma or TNF-alpha spleen cells, partially restored cell viability and caused a decrease in the levels of apoptosis in splenocytes from infected animals. Spleen cells from T. cruzi-infected mice had an apoptosis-specific pattern of internucleosomal DNA fragmentation which was most marked at the ninth day after infection when the plasma NO levels and parasitemia were increased. Treatment of infected mice with L-NMMA, anti-TNF-alpha, or anti-IFN-gamma mAbs caused reduction of both NO production and the amount of apoptotic cells, suggesting that NO plays a direct role in the induction of apoptosis in vivo. Taken together, these data support the hypothesis that, as well as modulating immunosuppression, NO produced by IFN-gamma and TNF-alpha activated macrophages plays a role in apoptosis induction during the acute phase of experimental T. cruzi infection.

Use of a simplified polymerase chain reaction procedure to detect Trypanosoma cruzi in blood samples from chronic chagasic patients in a rural endemic area.

The feasibility of using DNA amplification by the polymerase chain reaction (PCR) for specific detection of Trypanosoma cruzi in human blood specimens was investigated. One hundred blood samples were collected in an endemic area of Minas Gerais, Brazil. They were submitted to DNA extraction and PCR amplification with kinetoplast DNA-specific primers using a simplified boiling procedure that linearized most minicircle molecules without the aid of chemical reagents. Samples that gave negative results were checked for possible inhibition of amplification using primers derived from a human-specific sequence, and those showing some level of inhibition were retested after a new DNA extraction. Of 86 patients previously diagnosed as chagasic by serologic techniques, 83 were positive in our PCR test (sensitivity = 96.5%), including all the xenodiagnosis-positive patients and 21 (87.5%) of 24 xenodiagnosis-negative individuals. In addition, four of six patients with doubtful serologic results were confirmed as positive by PCR. Our results suggest that the PCR may be a useful complement to serology in the diagnosis of Chagas' disease, and that it is the most powerful technique available for parasite detection in patients with chronic disease.

High correlation between Chagas' disease serology and PCR-based detection of Trypanosoma cruzi kinetoplast DNA in Bolivian children living in an endemic area.

The detection of Trypanosoma cruzi kinetoplast DNA by polymerase chain reaction (PCR) amplification is a potentially powerful tool for the parasitological diagnosis of Chagas' disease. We have applied this technique in a field situation in Bolivia, where 45 children from a primary school were subjected to serological testing, buffy coat analysis and PCR diagnosis. 26 of the 28 serology-positive individuals were also positive by PCR. In addition, two serology-negative children gave a positive result by PCR, including one who was positive in the buffy coat test. These results suggest that PCR detection of T. cruzi DNA in blood can be a very useful complement to serology in Chagas' disease diagnosis in Bolivia.

Paleoparasitology of Chagas disease revaled by infected tissues from Chilean mummies.

Mummified tissues were sampled from bodies stored at the Museo Arqueologico de San Pedro de Atacama, northern Chile, dated from 2000 years BP-1400 AD, and Trypanosoma cruzi DNA was recovered using polymerase chain reaction (PCR) methodology. Amplification of the conserved region of the minicircle molecule of T. cruzi was achieved in four of the six samples tested. Amplified products corresponding to genetic fragments of the parasite were tested by hybridization experiments with positive results for T. cruzi specific molecular probe. The origin and dispersion of T. cruzi human infection is discussed as well as the molecular paleoparasitological approach, and what it may represent in an evolutionary perspective.

Anaemia, iron deficiency and malaria in a rural community in Brazilian Amazon.

OBJECTIVE: To assess the incidence of anaemia, iron deficiency and malaria in a malaria-endemic community. DESIGN: Three consecutive cross-sectional surveys (A, B and C) of the whole population made at 6-month intervals and malaria surveillance between the surveys. SETTING: Urupá, a rural community in Western Brazilian Amazon. SUBJECTS: 133 people of all age groups present in at least two cross-sectional surveys. INTERVENTIONS: Anaemic patients received ferrous sulphate during 3 months. Patients parasitized by intestinal nematodes were given mebendazole and parasitologically proven Plasmodium falciparum and P. vivax malaria attacks were treated with quinine or chloroquine plus primaquine. RESULTS: Anaemia (haemoglobin concentrations [Hb] below the cut-off values proposed by the World Health Organization) was diagnosed in respectively 10.0% (13 of 130) subjects in survey A, 9.2% (10 of 109) in B and 29.7% (27 of 91) in C. Depleted iron stores [serum ferritin (SF) < 12 micrograms/l] were detected in 10.0% subjects in survey A, 10.1% in B but in only 8.8% subjects in survey C. Concomitant anaemia and low SF was detected in 5.4% subjects in survey A, 3.7% in B and 6.6% in C. Mean Hb from anaemic patients diagnosed and treated during the study (n = 17) raised 1.2 g/dl after iron therapy and most of them (13 of 17, 76.5%) became non-anaemic. The highest malaria transmission was observed between surveys B and C. People who suffered at least one malaria attack during this period (27 of 63) were at a slightly greater risk of subsequent anaemia (odds ratio = 2.85, 95% confidence interval 0.81-10.28). CONCLUSIONS: Both malaria and iron deficiency could be considered as important causes of anaemia in this population. SPONSORSHIP: Supported by grants from the UNDP/World Bank/World Health Organization Special Programme for Research and Training in Tropical Diseases (no. 890245), the Ministére des Affaires Etrangeres, France, and from the Fundação de Amparo à Pesquisa do Estado de São Paulo (no. 92/1336-4). M.A.C. was supported by a doctoral fellowship from the Conselho Nacional de Desenvolvimento Científico e Tecnológico.

Structural studies on fucoidans from the brown seaweed Sargassum stenophyllum.

The brown seaweed Sargassum stenophyllum biosynthesizes two different sets of fucoidans. One of them is characterized by higher percentages of glucuronic acid and fewer sulfate groups, which are situated on different sugar units. alpha-L-Fucose was the major component but other sugars like beta-D-galactose, beta-D-mannose, alpha-D-glucuronic acid, alpha-D-glucose and beta-D-xylose were also in substantial amounts. Fucoidans from the other set contain small amounts of alpha-D-glucuronic acid and high percentages of sulfate groups, which are concentrated on the fucose residues, with only fucose and galactose as major components. Structural studies of one fucoidan from each set suggest that these products have a general basic structure that has a formal resemblance to that of the fucosylated chondroitin sulfates from the body wall of sea cucumbers, namely, a linear core (formed mainly by (1-->6)-beta-D-galactose and/or (1-->2)-beta-D-mannose units) with branched chains of 'fucans' (formed by (1-->3) and/or (1-->4)-alpha-L-fucose, (1-->4)-alpha-D-glucuronic acid, terminal beta-D-xylose and, sometimes, (1-->4)-alpha-D-glucose). In fucoidans from the second set, the 'core' is reduced to short galactan chains.

Serum concentrations of insulin-like growth factor-I and proteoglycan synthesis rates in young rats: a comparative study between the regional diet of São Paulo state and casein diets.

Insulin-like growth factor-I (IGF-I), also known as somatomedin-C, is an important mediator of growth regulation. Serum concentrations of IGF-I and proteoglycan synthesis rates in the tibial epiphysis, an estimate of the biological response to IGF-I in a target tissue, were compared in weanling Wistar rats fed ad libitum (group 1) and with 50% restriction (group 2) with the regional diet of São Paulo State (RDSPS--a mean diet consumed by low-income families with rice, beans, sugar, meat, milk, fruits and other vegetables) and in pair-fed animals fed with casein diets (groups 3 and 4). Data are reported as mean +/- SD for 8 rats in each group. Proteoglycan synthesis rates (cpm/mg) were significantly higher in rats fed with the RDSPS-based diet (groups 1 and 2: 210.8 +/- 58.8, 136.6 +/- 17.6) than in pair-fed animals fed with an 11% casein diet (groups 3 and 4: 62.9 +/- 11.6, 37.7 +/- 13.7) and in control animals fed ad libitum with a 20% casein diet (group 5: 58.1 +/- 22.7). Furthermore, these rates were higher in animals fed ad libitum than in those fed with the same diets but with 50% restriction. However, similar differences between groups 1 to 4 were not observed in serum concentrations (ng/100 microliters) of IGF-I (group 1: 44.1 +/- 7.1; group 2: 40.8 +/- 3.8; group 3: 46.0 +/- 3.6; group 4: 41.6 +/- 3.4, and group 5: 63.2 +/- 7.8). These results suggest that serum IGF-I levels are not reliable indicators of IGF-I status in this experimental model.

A laboratory cage for foster nursing newborn mice.

We describe a cage to be used for foster nursing in order to guarantee that original mother's colostrum is not ingested by the newborn mice. A common (30.5 cm x 19.5 cm x 12.0 cm) mouse cage was fitted with a wire net tray with a mesh (1 cm x 1 cm), which divides the cage into an upper and a lower compartment. Mice born to females placed in the upper compartment pass through the mesh and fall into the lower compartment, where another lactating female with one or two of its own pups are. Of a total of 28 newborn mice of C3H/Hc and Swiss strains, 23 were successfully fostered. Important observations are presented to show that this is a valuable alternative for foster studies without great suffering on the part of the female.

Effect of dietary iron on the course of Plasmodium berghei malaria in young rats.

Clinical and experimental evidence suggests that iron-deficient hosts are less susceptible to severe malaria and that iron supplementation aggravates infection. In the present study, 60 weanling Wistar rats were fed standard diets with different iron concentrations: 21 mg/kg (group 1), 45 mg/kg (group 2) and 113 mg/kg (group 3). Ferrous sulfate (FeSO4 x 7H2O) was added to the normal-iron and iron-supplemented diets (groups 2 and 3, respectively). Data are reported as mean +/- SEM. After 16 days of regimen, eight rats from each group were killed to measure serum iron concentration (SI) and transferrin saturation capacity (TSC). At this moment, rats from group 1 were underweight and their dietary intake was significantly lower than that of animals from the other groups. Severe iron deficiency (SI = 49.2 +/- 4.5 micrograms/100 ml and TSC = 8.3 +/- 0.7%) was observed in rats from group 1, while the animals from the other groups were iron-sufficient (group 2: SI = 186.5 +/- 28.5 micrograms/100 ml and TSC = 27.3 +/- 3.4%; group 3: SI = 137.3 +/- 18.2 micrograms/100 ml and TSC = 21.3 +/- 2.3%). Nine animals from each group were then infected with the malaria parasite Plasmodium berghei, whereas three animals from each group were used as noninfected controls. Parasitemias (% of infected red blood cells) peaked 7 days post-infection in animals from groups 2 and 3 (mean values of 2.4% and 1.7%, respectively), but in animals from group 1 parasitemias increased until the 9th day post-infection (mean at peak, 2.3%) and parasite clearance was significantly slower than in the other groups.(ABSTRACT TRUNCATED AT 250 WORDS)