Neutrophil extracellular traps characterize caseating granulomas.

Tuberculosis (TB) remains one of the top 10 causes of death worldwide and still poses a serious challenge to public health. Recent attention to neutrophils has uncovered unexplored areas demanding further investigation. Therefore, the aim of this study was to determine neutrophil activation and circulatory neutrophil extracellular trap (NET) formation in various types of TB. Sera from TB patients (n = 91) and healthy controls (NHD; n = 38) were analyzed for NE-DNA and MPO-DNA complexes, cell-free DNA (cfDNA), and protease activity (elastase). We show that these NET parameters were increased in TB sera. Importantly, NET formation and NE activity were elevated in TB patients with extensive tissue damage when compared to those with minor damage and in patients with relapse, compared to new cases. We discuss the importance of balancing NET formation to prevent tissue damage or even relapse and argue to analyze circulating NET parameters to monitor the risk of disease relapse. To investigate the tissues for NETs and to find the source of the circulating NET degradation products, we collected sections of granulomas in lung and lymph node biopsies. Samples from other diseases with granulomas, including sarcoidosis (SARC) and apical periodontitis (AP), served as controls. Whereas NET formation characterizes the caseating granulomas, both caseating and non-caseating granulomas harbor DNA with unusual conformation. As TB is associated with hypercoagulation and thromboembolism, we further imaged the pulmonary vessels of TB patients and detected vascular occlusions with neutrophil aggregates. This highlights the dual role of neutrophils in the pathology of TB.

Knowledge about relationship of diets with non-communicable disease and prospects for amelioration by consuming mushrooms in South-East Nigeria.

Background: Knowledge plays a key role in shaping people's attitudes and behaviors to adopt healthy lifestyles, which is important in any program of NCD prevention. There is no data about how much the general public in South-East Nigeria (SE) knows about how their diets may dispose or help them prevent NCDs. Aim: This study aims to assess knowledge available on the relationship between NCDs and diets/foods including mushrooms among the general public of SE Nigeria. Methods: Data was collected using the survey questionnaire and interview method in the five SE States: Abia, Anambra, Ebonyi, Enugu, and Imo States. Simple descriptive statistics were used to analyze data. Results: A total of 846 responses were received from 1000 questionnaires. Proportion of respondents who knew (p^yes) about a relationship between diets and NCDs was 51.5%. p^yes was significantly higher among males than females and increased with age and level of education of respondents. It also varied among the States of SE Nigeria. Knowledge scores based on a scale of 0-5 revealed that respondents knew more about foods, which can reduce (2.3 ± 0.102) e.g., fruits/vegetables, than can increase (1.9 ± 0.096) e.g., sugary/starchy foods, the risk of NCDs. Varying proportions of respondents in all States of the SE knew that mushroom consumption can ameliorate NCDs. Conclusion: This study reveals that half of population of SE Nigeria knows about the relationship between diets and NCDs. A significant proportion also knows that mushroom consumption can ameliorate NCDs suggesting prospects of its utilization in preventing NCDs.

Immuno-informatics design of a multimeric epitope peptide based vaccine targeting SARS-CoV-2 spike glycoprotein.

Developing an efficacious vaccine for SARS-CoV-2 infection is critical to stemming COVID-19 fatalities and providing the global community with immune protection. We have used a bioinformatic approach to aid in designing an epitope peptide-based vaccine against the spike protein of the virus. Five antigenic B cell epitopes with viable antigenicity and a total of 27 discontinuous B cell epitopes were mapped out structurally in the spike protein for antibody recognition. We identified eight CD8+ T cell 9-mers and 12 CD4+ T cell 14-15-mer as promising candidate epitopes putatively restricted by a large number of MHC I and II alleles, respectively. We used this information to construct an in silico chimeric peptide vaccine whose translational rate was highly expressed when cloned in pET28a (+) vector. With our In silico test, the vaccine construct was predicted to elicit high antigenicity and cell-mediated immunity when given as a homologous prime-boost, triggering of toll-like receptor 5 by the adjuvant linker. The vaccine was also characterized by an increase in IgM and IgG and an array of Th1 and Th2 cytokines. Upon in silico challenge with SARS-CoV-2, there was a decrease in antigen levels using our immune simulations. We, therefore, propose that potential vaccine designs consider this approach.

Attenuated Subcomponent Vaccine Design Targeting the SARS-CoV-2 Nucleocapsid Phosphoprotein RNA Binding Domain: In Silico Analysis.

The novel coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has previously never been identified with humans, thereby creating devastation in public health. The need for an effective vaccine to curb this pandemic cannot be overemphasized. In view of this, we designed a subcomponent antigenic peptide vaccine targeting the N-terminal (NT) and C-terminal (CT) RNA binding domains of the nucleocapsid protein that aid in viral replication. Promising antigenic B cell and T cell epitopes were predicted using computational pipelines. The peptides "RIRGGDGKMKDL" and "AFGRRGPEQTQGNFG" were the B cell linear epitopes with good antigenic index and nonallergenic property. Two CD8+ and Three CD4+ T cell epitopes were also selected considering their safe immunogenic profiling such as allergenicity, antigen level conservancy, antigenicity, peptide toxicity, and putative restrictions to a number of MHC-I and MHC-II alleles. With these selected epitopes, a nonallergenic chimeric peptide vaccine incapable of inducing a type II hypersensitivity reaction was constructed. The molecular interaction between the Toll-like receptor-5 (TLR5) which was triggered by the vaccine was analyzed by molecular docking and scrutinized using dynamics simulation. Finally, in silico cloning was performed to ensure the expression and translation efficiency of the vaccine, utilizing the pET-28a vector. This research, therefore, provides a guide for experimental investigation and validation.

Cryptococcus neoformans seropositivity and some haematological parameters in HIV seropositive subjects.

BACKGROUND: Cryptococcus neoformans is an opportunistic fungal pathogen that causes meningitis worldwide and may be fatal in immunocompromised subjects. In Nigeria, cases have been reported with prevalence between 4 and 13.1% in Human Immunodeficiency virus (HIV) patients depending on the study subjects. This study was designed to assess the prevalence of cryptococcosis, CD4+T cell counts and possible effect on haematological parameters in HIV seropositive subject in Nnewi, South-Eastern Nigeria. METHOD: A total of four hundred and twenty-nine (429) subjects were recruited for the study. Of these, two hundred and ninety (290) were HIV positive and one hundred and thirty-nine (139) were HIV seronegative subjects recruited from the voluntary counseling and testing (VCT) unit and HIV care clinic at Nnamdi Azikiwe University Teaching Hospital Nnewi, Anambra State, Nigeria. Their ages were between 18-80 years. One hundred and thirty nine (139) apparently healthy HIV seronegative subjects were recruited as controls. Blood samples were taken for C. neoformans by Antigen lateral flow assay (CrAgLFA), HIV testing, CD4+T cell, platelet and Full blood count (FBC). RESULTS: Our results show that of the two hundred and ninety (290) who were HIV positive subjects investigated for cryptococcosis, 4 (1.4%) tested positive for CrAg of whom 1(25%) were male and 3(75%) were female. All those with cryptococcosis had their CD4 count below 200 cells/µL, three of them were on ART and one was not. There were significant differences in the CD4 counts (P<0.05) between those infected and not infected with C. neoformans. None of the control group tested positive to cryptococcosis. CONCLUSION: Widespread use of anti-retroviral therapy may have reduced C. neoformans infection. However, the threat remains and there may be a possibility that women may be a more vulnerable population.

Evaluation of some haemostatic parameters in falciparum malaria and HIV co-infection.

BACKGROUND: Studies from sub-Saharan Africa where malaria is endemic have observed high incidences of malaria and HIV co-infection. It has long been accepted that malaria causes alterations in haemostatic parameters and that HIV is associated with a wide range of haematological changes. We assessed the effect of the overlap of these infections on routine haemostatic parameters. METHOD: The study involved 337 subjects grouped according to their HIV and malaria status: Group 1 'Asymptomatic HIV seropositive, Plasmodium falciparum positive' (n = 61); Group 2 'Asymptomatic HIV seropositive, P. falciparum negative' (n = 73); Group 3 'Symptomatic HIV seropositive, P. falciparum positive' (n = 49); Group 4 'Symptomatic HIV positive P. falciparum negative' (n = 56); Group 5 'Control HIV negative, P. falciparum positive' (n = 52) and Group 6 'Control HIV negative, P. falciparum negative' (n = 46). Blood samples were taken for HIV testing, diagnosis of falciparum malaria and malaria parasite density counts. Citrated samples were used within one hour of collection for prothrombin time (PT) and activated partial thromboplastin time (APTT). CD4+ T cell counts, platelet count and haematocrit (Hct) were also performed. RESULTS: Our results demonstrate greater alterations in APTT, PT and platelet count with prolongation of APTT, PT and lower platelet counts in HIV and malaria co-infection. In spite of this, the co-infected subjects with mild to moderate parasitaemia did not show a bleeding tendency; however, the risk is higher in severe malaria. CONCLUSION: These results suggest that co-infected subjects with severe malaria have a higher risk of bleeding and would require greater monitoring.