Development and characterization of a new monoclonal antibody against SARS-CoV-2 NSP12 (RdRp)

SARS-CoV-2 NSP12, the viral RNA-dependent RNA polymerase (RdRp), is required for viral replication and is a therapeutic target to treat COVID-19. To facilitate research on SARS-CoV-2 NSP12 protein, we developed a rat monoclonal antibody (CM12.1) against the NSP12 N-terminus that can facilitate functional studies. Immunoblotting and immunofluorescence assay (IFA) confirmed the specific detection of NSP12 protein by this antibody for cells overexpressing the protein. Although NSP12 is generated from the ORF1ab polyprotein, IFA of human autopsy COVID-19 lung samples revealed NSP12 expression in only a small fraction of lung cells including goblet, club-like, vascular endothelial cells, and a range of immune cells, despite wide-spread tissue expression of spike protein antigen. Similar studies using in vitro infection also generated scant protein detection in cells with established virus replication. These results suggest that NSP12 may have diminished steady-state expression or extensive posttranslation modifications that limit antibody reactivity during SARS-CoV-2 replication.     

Effect of different activations of silver nanoparticle irrigants on the elimination of Enterococcus faecalis

Clinical Oral Investigations - This study aimed to compare the efficacy of silver nanoparticles (AgNPs) irrigating solution alone and following activation with photon-induced photoacoustic...     

American cutaneous leishmaniasis in infancy and childhood

Infant and young child skin diseases are among the most common features of morbidity throughout the tropics. Because the skin is directly exposed to the environment, it is considerably affected by climatic and local conditions such as vectors and microorganisms, as in the case of leishmaniasis. In America the observed magnitude of cutaneous leishmaniasis in children has led to the study of increased risk of exposure of this group due to the possibility of peri‐ and intradomiciliary transmission. The present review pretends to make a concrete approach all through the broad and main figures of this parasitic disease, including the clinical, physiopathological, epidemiological, diagnostic, and therapeutic aspects, in order to be used as a practical source of reference for pediatricians leading with tropical cutaneous pathology in the region.     

Knowledge about contraceptives in a population 15 years or older in a southern Brazilian city

In Brazil, prevalence of contraceptive use is high, but incorrect and inappropriate use is frequent, suggesting limited knowledge about methods. This study thus evaluated the knowledge about the most widely used contraceptive methods, measured through a score (0-10) and related factors. A population-based cross-sectional study was performed, with 3,542 subjects ages 15 or older, residing in the urban area of Pelotas, Rio Grande do Sul State. The mean overall knowledge score was 4.65 (sd = 2.07): 5.02 (sd = 2.10) for women and 4.18 (sd = 1.92) for men. Lower age, higher schooling, a report of unwanted pregnancy, and any lifetime use of contraceptive methods (exclusive or combined) were associated with higher knowledge scores among men, while for women, the determinants of higher knowledge were higher age, living with a partner, higher schooling, higher socioeconomic status, not reporting a religion, and any lifetime use of contraceptive methods (exclusive or combined). Despite high prevalence of any lifetime contraceptive use (75.3%), knowledge is still limited about the most widely used methods, as well as about the menstrual cycle and fertile period.     

The influence of the hemodialysis treatment time under oxidative stress biomarkers in chronic renal failure patients

SUMMARY: Oxidative stress possibly helps to promote the progression and complication of chronic renal failure (CRF). Hemodialysis (HD) may aggravate oxidative stress. In addition long time of treatment may intensify the oxidative stress. Thus, the aim of this study was to evaluate the effect of prolonged HD treatment under parameters of the oxidative stress. METHODS: Plasmatic thiobarbituric acid reactive substances (TBARS), plasmatic malondialdehyde (MDA), blood delta-aminolevulinate dehydratase (ALA-D) activity, ALA-D reactivation index, and erythrocytic reduced glutathione (GSH) were measured into two different groups of HD patients: recent treatment (n=36; HD duration: 17.7+/-1.71 months), and long time of treatment (n=26; HD duration: 82.2+/-6.32 months), and in a control group (n=40). RESULTS: Plasmatic TBARS and MDA levels were both elevated in HD patients. However, only MDA levels had positive correlation with time of HD treatment. Blood ALA-D activity was decreased in HD patients. The ALA-D reactivation index showed increase in HD patients, and it had correlation with the time of HD treatment. Erythrocytic GSH levels were increased in HD patients. CONCLUSIONS: Our results indicated that MDA levels and ALA-D reactivation index may be the better biomarkers to evaluate chronic oxidative stress in comparison with others markers analyzed in this study.     

Butyrylcholinesterase activity is reduced in haemodialysis patients: Is there association with hyperhomocysteinemia and/or oxidative stress?

Objectives:To quantify serum butyrylcholinesterase activity in haemodialysis patients and to evaluate if the homocysteine levels and/or oxidative stress biomarkers have an effect on butyrylcholinesterase.Materials and methods:Blood samples were collected from patients and healthy subjects (control). The plasma homocysteine and TBARS levels; serum butyrylcholinesterase activity; blood δ aminolevulinic acid dehydratase (ALA-D) activity and methahaemoglobin were analyzed. The mortality of the patients was also evaluated after 3 years.Results:The homocysteine was increased and butyrylcholinesterase decreased compared to control (p < 0.05). TBARS and methahaemoglobin were increased and ALA-D decreased (p < 0.05). The following correlations were found: homocysteine with butyrylcholinesterase (? 0.44); methahaemoglobin (0.41); ALA-D (? 0.68); and TBARS (0.66). The partial correlation between homocysteine with butyrylcholinesterase, withdrawn the effect of TBARS, was ? 0.30; all p < 0.05. Moreover, it was observed that 22% of the patients died due to cardiovascular problems.Conclusion:Thus, our findings support a direct association between the reduction of butyrylcholinesterase by the increase of homocysteine and an indirect effect by increase in oxidative stress.     

Diffuse (anergic) cutaneous leishmaniasis responding to amphotericin B

American cutaneous leishmaniasis is an important endemic zoonotic disease in the New World that comprises a spectrum of clinical manifestations. Diffuse cutaneous leishmaniasis (DCL) is a rare form of the disease characterized by antigen‐specific immunodeficiency that often presents with multiple disfiguring non‐ulcerated confluent nodules or plaques that involve large areas of the skin, resembling lepromatous leprosy. Relapse is invariable in advanced stages, despite aggressive chemotherapy, and a plethora of drugs has been tested with unchanging results. We report on a severe an exceptional case that resolved after treatment with amphotericin B, a drug considered only mildly effective, and discuss the therapeutic approach to this disease.