Bioactive compounds and benefits of by-products of Amazon babassu oil production: potential for dietary supplement, biomedical and food applications.

Babassu coconut (Attalea speciosa syn. Orbignya phalerata) contains an oil-rich nut and is primarily found in South America's Amazon region. Future market researchers predict an increase in the babassu oil market from USD 227.7 million in 2022 to USD 347.0 million by 2032, and the yield of babassu oil from babassu-processed waste could reach 90%. Of these, mesocarp flour is an underrated by-product used only for animal feed purposes by local producers. This comprehensive review focuses on advances in knowledge and understanding of phytochemicals from babassu oil by-products considering the mechanisms of action - covering antioxidant, antimicrobial, antiparasitic, anti-inflammatory, antithrombotic, immunomodulatory, and anticancer effects. Babassu coconut fruit contains free fatty acids, (poly)phenols, phytosterols, and triterpenes. Pytochemicals, antiparasitic and antibacterial activities of babassu mesocarp flour were shown, but fungi and viruses can get more attention. Beyond its antioxidant capacity, babassu mesocarp flour showed potential as a dietary food supplement. Aqueous suspensions of mesocarp flour with a higher preference for cancer cells than normal cells and an antithrombotic effect were also identified, probably related to the antioxidant capacity of its secondary metabolites. Mesocarp flour, a starch-rich fraction, is promising for application as biodegradable packaging to improve the oxidative stability of foods. Finally, low-added value fractions can be considered bio-waste/co-products, and their phytochemicals may attract interest for applications in medicine and nutrition. Toxicological concerns, trends, and gaps are discussed for the future of foods and related sciences.

Ensuring vaccine safety: Case studies of falsified influenza vaccines detected in Brazil.

Falsifications related to health technologies-including vaccines-are a growing threat to patient safety and health systems on a global scale and can cause serious harm to the population (especially vulnerable groups). In Brazil, the manufacturing and spread of counterfeit medicines are prevented through joint actions between different government agencies. In this study, we analyzed three cases of influenza vaccines suspected of counterfeiting. The samples were seized by officials and received by the National Institute for Quality Control in Health (INCQS), the national quality control reference laboratory of the Ministry of Health of Brazil, in 2010, 2017, and 2020. We report the results of our analytical investigations and emphasize the importance of strengthening the partnerships between various national agencies. The seized samples were visually inspected, and their information was compared with that of genuine vaccines (as recorded in the INCQS database). The specific analytical tests were based on quality control tests for biological products. Our results confirmed that all seized samples were falsified. We emphasize the importance of fostering international and intra-national collaborations between various national agencies (such as drug regulatory authorities, official laboratories, customs departments, police forces, and civil society). As demonstrated here, such collaborative actions are essential for combating the release of falsified medical products, safeguarding public health, and strengthening health systems.

Verification of the rabies virus glycoprotein lower limit of immunogenicity by serological assay.

Rabies lethality is close to 100% and annually 15 million people receive post-exposure prophylaxis. Testing for vaccines against this zoonosis should ensure its quality. A standardized test by the National Institutes of Health (NIH) test, based on mice immunization and challenge, has been used to determine the potency of vaccine lots. It has several disadvantages, the main one being its significant variability. Several in vitro methods like an Enzyme-linked immunosorbent assay (ELISA) have been proposed based on the quality and quantity of glycoprotein (Glptn) of rabies virus, but may also present limitations such as low sensitivity, instability and imprecision. The estimate of immunogenicity based on neutralizing antibody titer (Nab) evaluated by a serological test (ST) such as the Modified Rapid Fluorescent Focus Inhibition Test (mRFFIT), is not yet efectively applied for human vaccine. Nevertheless, a Nab concentration can be used as a predictor of clinical efficacy of this product in vaccinated humans, so, that can be applied in estimating the vaccine potency. The aim of this study was to verify the lower limit of immunogenicity of the viral Glptn content in mice using mRFFIT. The lower Glptn content by ELISA able to induce Nab response was determined. The results were correlated and demonstrated that ST was able to determine the Glptn immunogenicity lower limit. Our findings suggest that a test based on rabies Nabs may represent an additional alternative for the evaluation of rabies vaccines.

Development and pre-validation of a quantitative multi-dose serological assay for potency testing of inactivated rabies vaccines for human use.

It is mandatory to ensure the quality of biological products used in the prevention of rabies, a zoonosis with nearly 100% lethality. Fifteen million people receive post-exposure prophylaxis yearly. The vaccine batches are assessed by the National Institutes of Health (NIH) test which has several disadvantages such as significant variability and animal welfare issues. The estimation of immunogenicity based on titration of neutralizing antibodies (NA) is not applied to the human vaccine yet. Despite this, a satisfactory concentration of NA (0.5 IU/ml) can be used as a predictor of the clinical efficacy and for estimating rabies vaccine potency. The objective of this study was to develop and pre-validate a Serological Potency Test (SPT) using the modified Rapid Fluorescent Focus Inhibition Test (mRFFIT) to determine the potency of rabies vaccines for human use, demonstrating its relevance and reliability. The results show good agreement between the potencies determined by the SPT and the NIH test. The assay was able to distinguish between potent and sub-potent lots of vaccines. The results demonstrated that SPT is a viable candidate for validation and inclusion in pharmacopeias as a reduction and refinement for the NIH test.

Characterization of strains of Neisseria meningitidis causing meningococcal meningitis in Mozambique, 2014: Implications for vaccination against meningococcal meningitis.

INTRODUCTION: In sub Saharan Africa, the epidemiology, including the distribution of serogroups of strains of N. meningitidis is poorly investigated in countries outside "the meningitis belt". This study was conducted with the aim to determine the distribution of serogroups of strains of N. meningitidis causing meningococcal meningitis in children and adults in Mozambique. METHODS: A total of 106 PCR confirmed Neisseria meningitidis Cerebrospinal Fluid (CSF) samples or isolates were obtained from the biobank of acute bacterial meningitis (ABM) surveillance being implemented by the National Institute of Health, at three central hospitals in Mozambique, from January to December 2014. Serogroups of N. meningitidis were determined using conventional PCR, targeting siaD gene for Neisseria meningitidis. Outer Membrane Proteins (OMP) Genotyping was performed by amplifying porA gene in nine samples. RESULTS: Of the 106 PCR confirmed Neisseria meningitidis samples, the most frequent serotype was A (50.0%, 53/106), followed by W/Y (18.9%, 20/106), C (8.5%, 9/106), X (7.5%, 8/106) and B (0.9%, 1/106). We found non-groupable strains in a total of 15 (14.2%) samples. PorA genotypes from nine strains showed expected patterns with the exception of two serogroup C strains with P1.19,15,36 and P1.19-36,15 and one serogroup X with P1.19,15,36, variants frequently associated to serogroup B. CONCLUSION: Our data shows that the number of cases of meningococcal meningitis routinely reported in central hospitals in Mozambique is significant and the most dominant serogroup is A. In conclusion, although serogroup A has almost been eliminated from the "meningitis belt", this serogroup remains a major concern in countries outside the belt such as Mozambique.

Interlaboratory study for the establishment of Brazilian Bothrops Reference Venom and Antivenom for potency evaluation of Bothrops antivenom.

A collaborative study was performed for the establishment of the 5th lot of Brazilian Bothrops Reference Venom and the 1st lot of Brazilian Bothrops Reference Antivenom. All Brazilian manufacturers of Antibothrops Immunoglobulins and the National Control Laboratory participated of the study. The declared potency of the 5th lot of the Bothrops Reference Venom is 40.29 µg/0.5 ml, and the potency of the 1st lot of Bothrops Reference Antivenom is 6.51 mg/ml. For the potency evaluation of Bothrops Reference Venom the inter assay precision (gCV) was 3.25% in lab 01; 3.51% in INCQS; 4.71% in lab 03 and 25.11% in lab 02, and the inter laboratory precision was 13.76%. The intra assay precision of Bothrops Reference Antivenom determinations was 4.38% in INCQS; 8.47% in lab 02; 10.51% in lab 03 and 20.05% in lab 01. The inter assay precision was 3.51% in INCQS; 9.65% in lab 02; 18.03% in lab 01 and 20.23% in lab 03. The inter laboratory precision was 15.85%. Despite the high number of invalid results (55.6% for the pharmacopoeial method and 69.4% for the proposed method) the parallel line assay, have better inter laboratorial precision (gCV = 16.62%) than the pharmacopoeial potency assay (gCV = 38.28%).

Validação de ensaio imunoenzimático utilizando-se o Conceito do Erro Total, os Perfis de Exatidão e o Índice de Exatidão como alternativa à abordagem clássica do ICH / Validation of an enzyme-linked immunosorbent assay by using the Total Error Concept, the Accuracy Profiles and the Accuracy Index as alternatives to the traditional ICH approach

O uso do conceito do Erro Total em validação de métodos analíticos é uma abordagem que incorpora a soma da veracidade e precisão. Este método utiliza ainda Perfis de Exatidão baseados em intervalos de tolerância para decidir se um modelo de calibração dará resultados de qualidade, e realiza o controle do risco de aceitar uma metodologia imprópria. Foram aplicados o Conceito do Erro Total, os perfis de Exatidão e o Índice de Exatidão na validação de um ensaio imunoenzimático (ELISA) para determinar o teor residual de ovoalbumina em vacinas. Este estudo testou uma amostra pura e fortificada com três diferentes concentrações (baixa, média e alta). A abordagem foi usada também para avaliar o método de cálculos que renderia resultados mais exatos. Os resultados obtidos no intervalo de concentrações testado com o modelo de cálculos escolhido foram: veracidade - ER % de 1,61 % a 12,15 %; precisão intermediária - CV% de 6,91 % a 9,31 % e o Erro Total de 9,83 % a 19,07 %, obtendo-se dados em conformidade com os guias internacionais. O estudo demonstrou que o método empregado é confiável para avaliar o teor de ovoalbumina, e que a abordagem do Conceito do Erro Total apresenta aplicabilidade na validação de ensaios imunoenzimáticos...

Avaliação comparativa da atividade biológica de heparinas não fracionadas pelas metodologias de inibição da coagulação do plasma ovino (ICPO) e de tempo de tromboplastina parcial ativada (TTPA) / Comparative study on the biological activity of non-fractionated heparin by using sheep plasma coagulation inhibition assay (SPCIA) and activated partial thromboplastin time (APTT) methodologies

O objetivo deste trabalho foi realizar o estudo comparativo entre os métodos ICPO e TTPA, para avaliar a eficácia da implantação do TTPA como método para a avaliação da segurança e eficácia de heparinas não fracionadas em produtos farmacêuticos. Foram avaliados, comparativamente, cinco lotes de diferentes fabricantes de heparinas não fracionadas (polissacarídeo sulfatado usado como droga anticoagulante), de origem suína ou bovina, testadas com base no 5º Padrão Internacional de Heparina. Esses produtos foram provenientes de coletas efetuadas pelas autoridades sanitárias para análise no Instituto Nacional de Controle de Qualidade em Saúde (INCQS). As amostras foram analisadas quanto à pureza e potência anticoagulante, por meio de duas metodologias: inibição da coagulação do plasma ovino (ICPO) e tempo de trombo plastina parcial ativada (TTPA). Houve boa concordância entre as duas metodologias, sendo que a técnica TTPA apresentou ser mais simples, rápida e objetiva, quando da utilização do coagulômetro para a medição do tempo de formação de coágulos, em detrimento da leitura subjetiva dos graus de coagulação no ensaio de ICPO. A implantação e a execução do TTPA em paralelo à utilização do ICPO garantirão o aumento de sensibilidade técnica na avaliação da segurança e eficácia de heparinas não fracionadas.

Multiplex-PCR serotyping of Listeria monocytogenes isolated from human clinical specimens

The genus Listeria is composed of six species of which Listeria monocytogenes is considered the single pathogenic species that causes listeriosis in humans. Of the 13 serovars of L. monocytogenes, 1/2a, 1/2b and 4b are responsible for the majority of clinical cases. The aim of this work was to detect L. monocytogenes in the cerebrospinal fluid sample of premature newborns and to characterize this sample using biotyping, serotyping and molecular typing. The results indicated the presence of L. monocytogenesin the clinical sample studied. Moreover, the isolate was identified as the 4b serovar that was characterized by the presence of a unique 691 bp band after analysis using the Multiplex-PCR technique. The results of repeated Multiplex-PCR and sequencing have indicated that the L. monocytogenes isolate was an atypical 4b serovar, which is the first time this finding has been reported.

Multiplex-PCR serotyping of Listeria monocytogenes isolated from human clinical specimens.

The genus Listeria is composed of six species of which Listeria monocytogenes is considered the single pathogenic species that causes listeriosis in humans. Of the 13 serovars of L. monocytogenes, 1/2a, 1/2b and 4b are responsible for the majority of clinical cases. The aim of this work was to detect L. monocytogenes in the cerebrospinal fluid sample of premature newborns and to characterize this sample using biotyping, serotyping and molecular typing. The results indicated the presence of L. monocytogenesin the clinical sample studied. Moreover, the isolate was identified as the 4b serovar that was characterized by the presence of a unique 691 bp band after analysis using the Multiplex-PCR technique. The results of repeated Multiplex-PCR and sequencing have indicated that the L. monocytogenes isolate was an atypical 4b serovar, which is the first time this finding has been reported.

Antimicrobial susceptibility of Haemophilus influenzae isolates collected from 4 centers in Brazil (1990-2003).

Antimicrobial susceptibility was determined for 174 Haemophilus influenzae strains collected from patients with infection before and after vaccination against Hib (1990-1999 and 2000-2003, respectively) from 4 public health -laboratories in 3 Brazilian states. All strains were characterized for serotype and beta-lactamase production and in vitro activity of the following antimicrobial agents: -ampicillin, amoxicillin/clavulanate, ceftriaxone, rifampin, chloramphenicol, and trimethoprim/sulfamethoxazole (TMP-SMX). Minimum inhibitory concentrations were determined according to the guidelines of the National Committee for Clinical Laboratory Standards. Overall, ampicillin resistance was observed in 29 strains (17%), all beta-lactamase producers. All isolates were susceptible to amoxicillin/clavulanate and ceftriaxone. The prevalence of TMP-SMX-resistant isolates increased from 32.6% in the period 1990-1999 to 65.8% during the period 2000-2003. Among these isolates, 10.0% and 12.5% were resistant to ampicillin and chloramphenicol, respectively. Resistance to rifampin was detected in 8.2% and 9.7% of the strains, in 2 periods, respectively. Continued surveillance is necessary to monitor trends with the H. influenzae disease in Brazil.