Impacts of Covid-19 interventions on air quality: evidence from Brazilian metropolitan regions.

The Covid-19 pandemic has negatively disrupted the way our economy and society functions. Nonetheless, there have also been some positive externalities of the pandemic on the environment. This paper aims to evaluate the concentration of nitrogen dioxide in Brazilian metropolitan regions after the policies adopted to confront Covid-19. In terms of methodological approach, the study employs cross-sectional quantitative analyses to compare the period of 36 days, i.e., 12 March to 16 April-before (in 2019) and after (in 2020) the pandemic declaration. The data were obtained from the Sentinel 5-P low-Earth polar satellite concerning Brazilian metropolitan regions (n = 24). Thorough spatial and statistical analyses were undertaken to identify the pre- and during pandemic nitrogen dioxide concentrations. Complementarily, Spearman's correlation test was performed with variables that impact air quality. The study results a fall in nitrogen dioxide concentration levels in 21 of the 24 metropolitan regions which was observed. The Spearman's correlation coefficient between the nitrogen dioxide variation and the vehicle density was 0.485, at a significance level of 0.05. With these findings in mind, the paper advocates that while the pandemic has a significant negative consequence on the health of population globally, a series of measures that result in a new social organization directly interfere in the reduction of air pollution that contributes to the quality of the air we breathe.

Interference of fixatives and fixation period on the morphologic analysis of ovarian preantral follicles.

Ovine ovarian fragments (3 × 3 × 1 mm) were fixed in neutral buffered formalin (NBF), Carnoy's solution (CAR), Davidson's solution (DAV), or paraformaldehyde (PFA) for 12 h or 24 h. After this fixation time, each fragment was prepared for histological analysis. Although fixative and fixation period did not affect follicular and stromal cells density, the percentages of morphologically normal primordial and primary follicles was affected by the fixative type and period of fixation. Paraformaldehyde was not indicated as a fixative for ovarian fragments. Formalin was a suitable fixative only when the period of fixation was 12 h, while Carnoy was efficient after a fixation period of 24 h. In conclusion, the most indicated fixative for the morphological evaluation of ovarian preantral follicles was DAV, regardless of the fixation period, that is 12 or 24 h.

Heterotopic autotransplantation of equine ovarian tissue using intramuscular versus subvulvar grafting sites: Preliminary results.

Ovarian tissue transplantation (OTT) is a technique well established and successfully applied in humans using mainly orthotopic or heterotopic transplantation sites. In livestock, OTT is still in its infancy and, therefore, different aspects of the technique, including the efficiency of different heterotopic OTT sites as well as the potential effect of age (i.e., young vs. old mares) in the ovarian graft quality, need to be investigated. The present study investigated the efficacy of the intramuscular (IM) or the novel subvulvar mucosa (SV) heterotopic autotransplantation sites to maintain the survivability of the grafts for 3 and 7 days post-OTT. Ovarian biopsy fragments were obtained in vivo and distributed to the following treatments: Fresh control group (ovarian fragments immediately fixed), SV-3, IM-3, SV-7, and IM-7. During and after graft harvesting, the macroscopic characteristics of the grafts (i.e., adherence, morphology, and bleeding) were scored, and the percentages of morphologically normal and developing preantral follicles as well as the follicular and stromal cell densities of the grafts were evaluated. The results were that similar (P > 0.05) macroscopic scores were observed between both transplantation sites 7 days post-OTT, with positive correlations (P < 0.01) found among adherence, morphology, and bleeding of the grafts. A lower (P < 0.05) percentage of morphologically normal follicles was found 7 days post-OTT in the SV site (82%) compared with the Fresh control group (99%) and IM site (95%); however, the percentages of developing follicles were similar (P > 0.05) between both transplantation sites 7 days post-OTT (30-43%). Although similar (P > 0.05) follicular densities were found in both transplantation sites in young and old mares at 3 and 7 days post-OTT, large individual variation in the follicular depletion rate was observed regardless of transplantation site. The Fresh control group and SV-7 treatments had higher (P < 0.05) stromal cell densities in young and old mares compared with both IM-7 treatments. When comparing transplant sites between young and old mares, the follicular density in old mares and the stromal cell density in young mares were greater (P < 0.05) in the SV than in the IM site. In conclusion, even though the transplantation sites differentially affected some end points, overall comparable findings of the OTT technique using both heterotopic autotransplantation sites (i.e., IM and SV) for equine ovarian tissue were observed.

Toxicological in vitro and subchronic evaluation of LASSBio-596.

LASSBio-596, 2-[4-(1,4-tiazinan-4-ylsulfonyl) phenylcarbamoyl] benzoic acid, is an achiral compound containing a subunit carboxylic amide, was capable of preventing induced mechanical and morphological changes in the lungs that commonly caused the onset of asthma. Previous studies to determine the acute toxicity of oral LASSBio-596 at dose of 2000mg/kg caused no deaths in any of the tested animals. To further evaluate the safety of LASSBio-596, in vitro and in vivo tests were carried out. Regarding to in vitro test were used renal, hepatic, pulmonary, cardiac, neurologic and intestinal cell lines. They were evaluated using neutral red (NR) and [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assays. Micronuclei also was performed. Concerning to in vivo was performed subchronic on Wistar rats at doses of 10, 50, and 250mg/kg and zebrafish test. The in vitro tests results showed the safety of LASSBio-596. However, subchronic toxicity study results revealed changes in the blood parameters of amylase, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose and creatine kinase (CK) which is used for cardiotoxicity evaluation, although, did not identify any histopathological alterations. However, zebrafish test demonstrated cardiac damage. It was impossible to estimate the no-observed-adverse-effect-levels and lowest observed-adverse-effect level due to the presence of cardiotoxicity in all tested doses.

Evaluation of the biodegradability of petroleum in microcosm systems by using mangrove sediments from Camamu Bay, Bahia, Brazil.

We investigated the biodegradability of oil in mangrove sediment from Camamu Bay and measured its effect on the bacterial community. Microcosms of mangrove sediment were contaminated with 0.1, 0.5, 1, 2, and 5% (w/v) oil, and the microbial activity was compared to that in uncontaminated sediment. The evolution of CO2 and gas chromatography showed the mineralization of oil compounds, which could reach 100%. Bacterial diversity was determined by polymerase chain reaction using a set of primers for the V3 and V6-V8 regions of 16S rDNA. The band profile obtained by denaturing gradient gel electrophoresis of the amplicons that were obtained for the V3 region showed a negative correlation between band number and oil concentration, whereas that of the V6-V8 region showed a positive correlation between band numbers and oil concentration. The latter also gave similar results for microcosms that were contaminated with 2 and 5% oil. These results demonstrate the mangrove sediment's capacity to recover from oil contamination (in vitro) and suggest that native mangrove microorganisms contain enzymes necessary for the catabolism of oil.

Construction and validation of metagenomic DNA libraries from landfarm soil microorganisms.

Landfarming biodegradation is a strategy used by the petrochemical industry to reduce pollutants in petroleum-contaminated soil. We constructed 2 metagenomic libraries from landfarming soil in order to determine the pathway used for mineralization of benzene and to examine protein expression of the bacteria in these soils. The DNA of landfarm soil, collected from Ilhéus, BA, Brazil, was extracted and a metagenomic library was constructed with the Copy Control(TM) Fosmid Library Production Kit, which clones 25-45-kb DNA fragments. The clones were selected for their ability to express enzymes capable of cleaving aromatic compounds. These clones were grown in Luria-Bertani broth plus L-arabinose, benzene, and chloramphenicol as induction substances; they were tested for activity in the catechol cleavage pathway, an intermediate step in benzene degradation. Nine clones were positive for ortho-cleavage and one was positive for meta-cleavage. Protein band patterns determined by SDS-polyacrylamide gel electrophoresis differed in bacteria grown on induced versus non-induced media (Luria-Bertani broth). We concluded that the DNA of landfarm soil is an important source of genes involved in mineralization of xenobiotic compounds, which are common in gasoline and oil spills. Metagenomic library allows identification of non-culturable microorganisms that have potential in the bioremediation of contaminated sites.

Application of denaturing gradient gel electrophoresis for detection of bacterial and yeast communities along a salinity gradient in the estuary of the Cachoeira River in Brazil.

An estuary is a transition zone between freshwater and marine ecosystems, resulting in dilution of seawater. Estuaries are also considered environments of intense biological activity related to the processes of nutrient cycling. The aim of this study was to evaluate the microbial community composition along a salinity gradient in the estuary of the Cachoeira River, located in southern Bahia, Brazil. The analysis of bacterial and yeast communities was performed by determining the denaturing gradient gel electrophoresis band richness. Formation of zones with similar profiles of bands was observed, and the increasing richness at the intermediate zone demonstrated a clear spatial distinction of communities depending on salinity. In addition, the dissolved oxygen content, temperature, pH, salinity, and dissolved inorganic nutrient contents (NH3(+), NO2(-), NO3(-), PO4(-)) were determined. Nutrients were distributed in similar patterns, with decreasing concentrations as the salinity increases.

Detection by denaturing gradient gel electrophoresis of ammonia-oxidizing bacteria in microcosms of crude oil-contaminated mangrove sediments.

Currently, the effect of crude oil on ammonia-oxidizing bacterium communities from mangrove sediments is little understood. We studied the diversity of ammonia-oxidizing bacteria in mangrove microcosm experiments using mangrove sediments contaminated with 0.1, 0.5, 1, 2, and 5% crude oil as well as non-contaminated control and landfarm soil from near an oil refinery in Camamu Bay in Bahia, Brazil. The evolution of CO(2) production in all crude oil-contaminated microcosms showed potential for mineralization. Cluster analysis of denaturing gradient gel electrophoresis-derived samples generated with primers for gene amoA, which encodes the functional enzyme ammonia monooxygenase, showed differences in the sample contaminated with 5% compared to the other samples. Principal component analysis showed divergence of the non-contaminated samples from the 5% crude oil-contaminated sediment. A Venn diagram generated from the banding pattern of PCR-denaturing gradient gel electrophoresis was used to look for operational taxonomic units (OTUs) in common. Eight OTUs were found in non-contaminated sediments and in samples contaminated with 0.5, 1, or 2% crude oil. A Jaccard similarity index of 50% was found for samples contaminated with 0.1, 0.5, 1, and 2% crude oil. This is the first study that focuses on the impact of crude oil on the ammonia-oxidizing bacterium community in mangrove sediments from Camamu Bay.

Rapid sample preparation procedure for As speciation in food samples by LC-ICP-MS.

This paper describes a rapid method for arsenic (As) speciation by LC-ICP-MS in several types of food samples. Prior to analysis, samples were milled and the As species extracted from biological tissues by sonication in only 2 min with a solution containing MeOH (10%, v/v) plus HNO3 (2%, v/v). As species were separated by LC using an anion exchange column. Method detection limits for AsB, As³âº, DMA, MMA and As5⁺ were 1.3, 0.9, 0.6, 0.7 and 0.8 ng g⁻¹, respectively. Method accuracy and precision were traceable to Certified Reference Materials SRM1577 bovine liver from the National Institute of Standards and Technology, CE278 mussel tissue from the Institute of Reference Materials and Measurements and DOLT-3 dogfish liver tissue and DORM-3 fish protein from the National Research Council of Canada. Finally, the method was applied to speciate As in food samples (egg, fish muscle, beef and chicken) purchased in Brazilian markets.

PP175. Gestational hypertensive syndromes: Risk factors identification and complications in a primiparous population.

INTRODUCTION: Preeclampsia is a major cause of maternal and perinatal morbidity and mortality in Brazil and worldwide. It is a heterogeneous and multifactorial disorder, with no clear etiology and pathophysiology. The identification of risk factors for its development may help in prevention and early diagnosis of disease clinical onset. OBJECTIVES: To identify risk factors and complications of pre-eclampsia (PE) and gestational hypertension (GH) in a population of primiparas, who had birth care at the Ceilandia Regional Hospital of Brasilia - Federal District. METHODS: Case-control study was conducted from November 2009 to December 2010 in a population of primiparas. We included all primiparas diagnosed with PE/GH, with random choice of normotensive primiparas in a ratio of 1:3. The variables studied as socio-demographic, obstetric, maternal and perinatal complications were collected using pre-tested questionnaire, after informed consent of the pregnant women. Chi-square and Fisher's multiple comparison were used in the data analysis, considering results significant when p <0.05. RESULTS: Among 1264 primiparas analyzed, 972 (76.9%) were normotensive and 292 (23.1%) hypertensive patients, being 64 (22%) with gestational hypertension and 228 (78%) with PE. Risk factors such as maternal age over 35years, women whose mother and/or sister with chronic hypertension history or who developed hypertension during pregnancy, use of male condom as a contraceptive method, obesity and excessive weight gain during pregnancy showed statistically significant correlation with development of the pathology. CONCLUSION: The development of gestational hypertensive syndrome in primiparous women is associated with family history and obesity, factors that can be used as screening for early identification of patients at risk of PE development.

Heparanase expression and activity influences chondrogenic and osteogenic processes during endochondral bone formation.

Endochondral bone formation is a highly orchestrated process involving coordination among cell-cell, cell-matrix and growth factor signaling that eventually results in the production of mineralized bone from a cartilage template. Chondrogenic and osteogenic differentiation occur in sequence during this process, and the temporospatial patterning clearly requires the activities of heparin binding growth factors and their receptors. Heparanase (HPSE) plays a role in osteogenesis, but the mechanism by which it does so is incompletely understood. We used a combination of ex vivo and in vitro approaches and a well described HPSE inhibitor, PI-88 to study HPSE in endochondral bone formation. In situ hybridization and immunolocalization with HPSE antibodies revealed that HPSE is expressed in the peri-chondrium, peri-osteum, and at the chondro-osseous junction, all sites of key signaling events and tissue morphogenesis. Transcripts encoding Hpse also were observed in the pre-hypertrophic zone. Addition of PI-88 to metatarsals in organ culture reduced growth and suggested that HPSE activity aids the transition from chondrogenic to osteogenic processes in growth of long bones. To study this, we used high density cultures of ATDC5 pre-chondrogenic cells grown under conditions favoring chondrogenesis or osteogenesis. Under chondrogenic conditions, HPSE/Hpse was expressed at high levels during the mid-culture period, at the onset of terminal chondrogenesis. PI-88 addition reduced chondrogenesis and accelerated osteogenesis, including a dramatic up-regulation of osteocalcin levels. In normal growth medium, addition of PI-88 reduced migration of ATDC-5 cells, suggesting that HPSE facilitates cartilage replacement by bone at the chondro-osseous junction by removing the HS component of proteoglycans, such as perlecan/HSPG2, that otherwise prevent osteogenic cells from remodeling hypertrophic cartilage.

Enzootiology of Trypanosoma evansi in Pantanal, Brazil.

In order to better understand the enzootiology of trypanosomiasis caused by Trypanosoma evansi in the Brazilian Pantanal we examined domestic and wild mammals by microhematocrit centrifuge technique (MHCT), immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). T. evansi infection was detected in all species sampled with exception of the sheep and the feral pig. High parasitemias were observed in capybaras (5/24), coatis (18/115), horses (31/321) and dogs (3/112). Among these species, only the capybaras did not develop anemia. Low parasitemias, only detected by PCR, were found in buffaloes (18/43), bovines (29/331), marsupials (1/4), small rodents (14/67), bats (7/18), and one armadillo (1/8). The highest prevalence of T. evansi infection was recorded in horses (73%), although no neurological signs in infected horses were observed. Diagnosis through standard parasitological tests and IFAT should be used with caution since they may overlook comprovedly infected horses. The relationship between ranch management and T. evansi infection in horse was investigated. The importance of other transmission mechanisms apart from the tabanids and reservoir hosts are discussed.

Using PCR for unraveling the cryptic epizootiology of livestock trypanosomosis in the Pantanal, Brazil.

Trypanosoma vivax and Trypanosoma evansi are livestock parasites of economic importance in Africa, Asia and South America. In the Pantanal, Brazil, they cause economic losses in both cattle and equines. Little is known of their maintenance and spread in nature, particularly in terms of reservoirs and means of mechanical transmission. Here we report for the first time the use of PCR for the detection of T. vivax and T. evansi in bovines, buffaloes and sheep. Whereas parasitological diagnosis detected only two T. vivax infections, one in buffalo and another in a cow, PCR detected infections in 34.8% buffaloes, 44.7% bovines and 37.3% sheep. Trypanozoon primers detected 41.8% infections in buffaloes and 8.1% in cattle. PCR revealed 6.9% mixed infections in buffaloes and 5.3% in cattle. The potential role of cattle and buffaloes as hosts and reservoirs of T. vivax is discussed, as well as the implications of possible extravascular foci in the maintenance of livestock trypanosomosis.

[Triatoma infestans in area under entomological surveillance for Chagas' disease in São Paulo State, Brazil]. / Triatoma infestans em área sob vigilância entomológica para doença de Chagas, Estado de São Paulo, Brasil.

In response to notification of the capture of a winged triatomine bug by local inhabitants, an epidemiological investigation was investigated in the rural area of the Municipality of Paulínia, São Paulo State (Brazil). This led to the collection of 109 specimens of Triatoma infestans from peridomiciliary breeding sites. Local conditions favored colonization by triatomine bugs: many abandoned outbuildings were inhabited by pigeons and sparrows, which meant abundant feed for these bugs. Thus, surveillance is mandatory as long as conditions for triatomine breeding persist. Nevertheless, entomological and serological indicators ruled out immediate concern over vectorial transmission of Chagas' disease in this municipality. All domiciliary units of the region were sprayed with residual-effect insecticides and the triatomine breeding sites were destroyed by the use of mechanical control of bird nests. The importance of reporting the occurrence of triatomine bugs, particularly Triatoma infestans, in this region, is emphasized.

Influence of menstrual cycle on NK activity.

Natural killer (NK) cells are CD3- CD56+ and/or CD16+ cytotoxic lymphocytes that mediate first-line defense against various types of target cells without prior immunization. To assess the effect of the menstrual cycle and gender on NK activity we evaluated 30 healthy women (mean age 28.1 years, range 21-39) in follicular and luteal phases, 29 postmenopausal women (mean age 58.8 years, range 42-72) and 48 healthy men (mean age 31.6 years, range 21-40). In a flow cytometric test of NK activity, peripheral blood mononuclear effector cells were mixed with K562 targets cells labeled with DiO (3,3'-dioctadecyloxacarbocyanine perchlorate) at effector:target cell ratios of 40, 20, 10 and 5:1. Dead cells were stained with propidium iodide and results were expressed as lytic units per 10(7) cells. In addition, progesterone levels were determined in the luteal phase of the menstrual cycle of healthy women by a chemiluminescence assay. Our results showed that (1) NK cytotoxicity was higher in the follicular than in the luteal phase of the menstrual cycle (P < 0.0001); (2) postmenopausal women and men showed NK activity similar to women in the follicular phase but higher than women in the luteal phase of the menstrual cycle (P < 0.05); and (3) there was no correlation between NK activity and levels of progesterone. The data suggest that progesterone does not influence NK activity directly and that other factors may explain the reduction of NK activity in the luteal phase of the menstrual cycle.

[Malaria in the region of Campinas, São Paulo, Brazil, 1980 to 1994]. / Malária na região de Campinas, São Paulo, Brasil, 1980 a 1994.

According to epidemiological data, from 1980 to 1994, 2,781 cases of malaria were registered as follows: DIR XII-Campinas (49.3%), DIR XV-Piracicaba (41.3%) and DIR XX-São João da Boa Vista (9.4%). The Plasmodium vivax was found in 70.6% of the patients; Plasmodium falciparum in 25.4% and 4% of the patients presented a mixed infection. The epidemiological classification showed that 95% of the cases came from the States of Rondônia, Mato Grosso and Par. The masculine sex, between the age of 20 to 39 years old, was responsible for 84.3% of the confirmed cases. During the study period, 9 cases of induced malaria were registered: 5 of which through blood transfusion, 3 through the sharing of syringes and infected needles among drug users and one occurrence of congenital malaria. 5 deaths were registered in patients prime-infected by Plasmodium falciparum with late diagnosis. The set of variables studied enabled a greater understanding of the epidemiology of malaria in the region and to subsidize and indicate the process of decentralization of attendance, diagnosis and treatment of the patient, as well as the control and epidemiological surveillance of the endemic disease in Campinas region and in the State of São Paulo.

Factor V leiden and factor II G20210A mutations in patients with recurrent abortion.

Recurrent abortion (RA) represents an intriguing problem in obstetric practice in which genetic and acquired factors may play a role. In the present investigation we sought to assess the possibility that inherited thrombophilia might determine the risk of RA. We therefore investigated the prevalence of two genetic abnormalities frequently associated with venous thrombosis [factor V Leiden (FVL) and factor II G20210A] in 56 patients with primary or secondary abortion and in 384 healthy control women. Polymerase chain reaction amplification followed by digestion with the restriction enzymes MnlI and HindIII was used to define the FVL and FII G20210A genotypes respectively. FVL was found in 4/56 patients (7.1%) and in 6/384 controls (1.6%), yielding an odds ratio (OR) for RA related to FVL of 4.9 [95% confidence interval (CI): 1.3-17.8]. FII G20210A was detected in 2/56 (3.6%) patients and in 4/384 (1%) controls (OR for RA: 3.5, CI: 0.6-19.7). In conclusion, FVL and FII G20210A mutations in patients with RA were more prevalent in comparison with controls. These data support a role for both mutations as determinants of the risk of RA and strengthen the notion that thrombophilia plays a role in this clinical entity.

Hematology of natural bovine trypanosomosis in the Brazilian Pantanal and Bolivian wetlands.

We report hematological changes observed in natural cases of bovine trypanosomosis due to Trypanosoma vivax in beef and dairy cattle from Bolivian wetlands and Pantanal, Brazil. The main hematologic changes produced by T. vivax infections were anemia and severe leucopenia. The cattle presented macrocytic hypochromic anemia. The leukocyte changes were characterized by relative lymphocytosis and monocytosis and decrease in the neutrophil counts. The clinical signs were lachrymation, progressive weakness, marked weight loss, inappetence, diarrhea and abortions during the third trimester of pregnancy.

The seroprevalence of equine trypanosomosis in the pantanal.

Since little information is available on the epizootiological status of Trypanosoma evansi in South America and particularly Brazil, we evaluated equine serum samples collected in 1993, 1994, 1995 and 1997 for the presence of antibodies against this trypanosome species. Our study shows corroborative evidence about the correlation among high T. evansi seroprevalence and the rainy season in the Pantanal, Brazil. The higher seroprevalence was 79.2% in horses from a ranch located in the Nhecolândia sub-region in 1994 and the lower 5.8% in animals from the same ranch in 1997. No seroprevalence was found in 1993. The possible re-introduction of T. evansi in the region as well as the relationship among our results with the outbreaks reported in 1994, are briefly discussed.

Leishmaniose tegumentar americana na regiäo nordeste do Estado de Säo Paulo - Brasil / American tegumentary leishmaniasis in the norheastern state of Säo Paulo - Brasil

Os autores descrevem um surto de leishmaniose tegumentar americana (LTA) instalado na regiao nordeste do Estado de Sao Paulo em 1992. Após a notificaçao de doze casos humanos em uma área rural do município de Itupeva, foi realizado um inquérito epidemiológico, destacando-se 34 por cento de positividade da populaçao à Reaçao Intradérmica de Montenegro. Constatou-se uma grande diversidade da fauna flebotomínica local, com predominância de L. migonei, L. intermedia e L. whitmani, tanto no domicílio, quanto na margem da mata. Ressalta-se a presença na margem da mata de L. longipalpis.