Generation and characterization of avian IgY antibodies for detecting beta-casein A1 in bovine milk.

Beta-casein is a primary milk protein that constitutes approximately 30% of the casein in bovine milk, with the two most common types in cattle being A1 and A2. The A2 protein differs from the A1 version due to a mutation in the codon at position 67, resulting in a histidine to proline substitution. However, the bioactive peptide, beta-casomorphine-7 (BCM7), which originates from partial proteolysis of the A1 variant, has been linked to several gastrointestinal disorders in humans. Production of A1 beta casein-free products is increasing demand in the milk market, worldwide. This study generated and characterized a polyclonal IgY antibody that specifically recognizes the A1 beta-casein protein present in cow's milk. A commercially available IgY anti-A1 antibody was used as a positive control, and the sensitivity and specificity of both the commercial and produced anti-A1 antibodies were evaluated. The results showed 100% sensitivity and specificity of 100% of the commercial IgY anti-A1. The in-house produced anti-A1 antibody demonstrated a sensitivity of 95.2% and a specificity of 100%, indicating its potential as a reliable and cost effective tool for detecting A1 beta-casein protein in milk samples.

In Vitro Diagnostic Assay to Detect SARS-CoV-2-Neutralizing Antibody in Patient Sera Using Engineered ACE-2 Mini-Protein.

The recent development and mass administration of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) vaccines allowed for disease control, reducing hospitalizations and mortality. Most of these vaccines target the SARS-CoV-2 Spike (S) protein antigens, culminating with the production of neutralizing antibodies (NAbs) that disrupt the attachment of the virus to ACE2 receptors on the host cells. However, several studies demonstrated that the NAbs typically rise within a few weeks after vaccination but quickly reduce months later. Thus, multiple booster administration is recommended, leading to vaccination hesitancy in many populations. Detecting serum anti-SARS-CoV-2 NAbs can instruct patients and healthcare providers on correct booster strategies. Several in vitro diagnostics kits are available; however, their high cost impairs the mass NAbs diagnostic testing. Recently, we engineered an ACE2 mimetic that interacts with the Receptor Binding Domain (RBD) of the SARS-2 S protein. Here we present the use of this engineered mini-protein (p-deface2 mut) to develop a detection assay to measure NAbs in patient sera using a competitive ELISA assay. Serum samples from twenty-one patients were tested. Nine samples (42.8%) tested positive, and twelve (57.1%) tested negative for neutralizing sera. The data correlated with the result from the standard commercial assay that uses human ACE2 protein. This confirmed that p-deface2 mut could replace human ACE2 in ELISA assays. Using bacterially expressed p-deface2 mut protein is cost-effective and may allow mass SARS-CoV-2 NAbs detection, especially in low-income countries where economical diagnostic testing is crucial. Such information will help providers decide when a booster is required, reducing risks of reinfection and preventing the administration before it is medically necessary.

Insights into Glucose-6-phosphate Allosteric Activation of ß-Glucosidase A.

Second-generation ethanol production involves the use of agricultural and forestry waste as feedstock, being an alternative to the first-generation technology as it relies on low-cost abundant residues and does not affect food agriculture. However, the success of second-generation biorefineries relies on energetically efficient processes and effective enzyme cocktails to convert cellulose into fermentable sugars. ß-glucosidases catalyze the last step on the enzymatic hydrolysis of cellulose; however, they are often inhibited by glucose. Previous studies demonstrated that glucose-6-phosphate (G6P) is a positive allosteric modulator of Bacillus polymyxa ß-glucosidase A, improving enzymatic efficiency, providing thermoresistance, and imparting glucose tolerance. However, the precise molecular details of G6P-ß-glucosidase A interactions have not yet been described so far. We investigated the molecular details of G6P binding into B. polymyxa ß-glucosidase A through in silico docking using the site identification by ligand competitive saturation technology followed by site-directed mutagenesis studies, from which an allosteric binding site for G6P was identified. In addition, a mechanistic shift toward the transglycosylation reaction as opposed to hydrolysis was observed in the presence of G6P, suggesting a new role of G6P allosteric modulation of the catalytic activity of ß-glucosidase A.

Salivary ß-glucosidase as a direct factor influencing the occurrence of halitosis.

ß-Glucosidases are enzymes present in all living organisms, playing a pivotal role in diverse biological processes. These enzymes cleave ß-glycosidic bonds between carbohydrates, or between a carbohydrate and a non-carbohydrate moiety, which may result in the liberation of volatile aglycones. Released compounds execute diverse physiological roles, while the industry takes advantage of exogenously added ß-glucosidases for aroma enrichment during food and beverage production. ß-Glucosidase enzymatic activity has been reported in human saliva and given the fact that these enzymes are involved in aroma release, we investigated here the correlation between ß-glucosidase activity in human saliva and the occurrence of halitosis. Measurement of salivary enzyme activity of 48 volunteers was performed using p-nitrophenyl-ß-d-glucopyranoside as substrate. Each volunteer was clinically evaluated by a dental surgeon and clinical and laboratorial data were statistically analyzed. Gas-chromatography of saliva headspace allowed the analysis of the direct role of exogenous ß-glucosidase on aromatic /volatile profile of saliva samples. The data demonstrated a positive correlation between halitosis and enzymatic activity, suggesting that the enzyme exerts a direct role in the occurrence of bad breath. Gas-chromatography analysis demonstrated that exogenously added enzyme led to the alteration of volatile organic content, confirming a direct contribution of ß-glucosidase activity on saliva volatile compounds release. Although halitosis is a multifactorial condition, the complete understanding of all governing factors may allow the development of more effective treatment strategies. Such studies may pave the way to the use of ß-glucosidase inhibitors for halitosis clinical management.

Using natural biomass microorganisms for drinking water denitrification.

Among the methods that are studied to eliminate nitrate from drinking water, biological denitrification is an attractive strategy. Although several studies report the use of denitrifying bacteria for nitrate removal, they usually involve the use of sewage sludge as biomass to obtain the microbiota. In the present study, denitrifying bacteria was isolated from bamboo, and variable parameters were controlled focusing on optimal bacterial performance followed by physicochemical analysis of water adequacy. In this way, bamboo was used as a source of denitrifying microorganisms, using either Immobilized Microorganisms (IM) or Suspended Microorganisms (SM) for nitrate removal. Denitrification parameters optimization was carried out by analysis of denitrification at different pH values, temperature, nitrate concentrations, carbon sources as well as different C/N ratios. In addition, operational stability and denitrification kinetics were evaluated. Microorganisms present in the biomass responsible for denitrification were identified as Proteus mirabilis. The denitrified water was submitted to physicochemical treatment such as coagulation and flocculation to adjust to the parameters of color and turbidity to drinking water standards. Denitrification using IM occurred with 73% efficiency in the absence of an external carbon source. The use of SM provided superior denitrification efficiency using ethanol (96.46%), glucose (98.58%) or glycerol (98.5%) as carbon source. The evaluation of the operational stability allowed 12 cycles of biomass reuse using the IM and 9 cycles using the SM. After physical-chemical treatment, only SM denitrified water remained within drinking water standards parameters of color and turbidity.

Improved enzymatic performance of graphene-immobilized ß-glucosidase A in the presence of glucose-6-phosphate.

Optimization of cellulose enzymatic hydrolysis is crucial for cost-effective bioethanol production from lignocellulosic biomass. Enzyme immobilization in solid support allows enzyme recycling for reuse, lowering hydrolysis costs. Graphene is a nanomaterial isolated in 2004, which possesses exceptional properties for biomolecule immobilization. This study evaluates the potential for ß-glucosidase recycling by immobilization on graphene nanosheets. Data reported here demonstrated that graphene-immobilized ß-glucosidase can be recycled for at least eight cycles. Immobilization did not change the optimal temperature of catalysis and improved enzymatic stability upon storage. The role of glucose-6-phosphate on immobilized enzyme was also investigated, demonstrating that glucose-6-phosphate acts as a mixed-type activator and improves storage stability of immobilized enzyme. Complete cellulose hydrolysis using graphene-immobilized ß-glucosidase in the presence of glucose-6-phosphate resulted in greatly improved hydrolysis rates, demonstrating the potential of this strategy for biomass hydrolysis.

Kinetic and biochemical characterization of Trypanosoma evansi nucleoside triphosphate diphosphohydrolase.

Nucleoside triphosphate diphospho-hydrolases (NTPDases) catalyze the hydrolysis of several nucleosides tri and diphosphate playing major roles in eukaryotes including purinergic signaling, inflammation, hemostasis, purine salvage and host-pathogen interactions. These enzymes have been recently described in parasites where several evidences indicated their involvement in virulence and infection. Here, we have investigated the presence of NTPDase in the genome of Trypanosoma evansi. Based on the genomic sequence from Trypanosoma brucei, we have amplified an 1812 gene fragment corresponding to the T. evansi NTPDase gene. The protein was expressed in the soluble form and purified to homogeneity and enzymatic assays were performed confirming the enzyme identity. Kinetic parameters and substrate specificity were determined. The dependence of cations on enzymatic activity was investigated indicating the enzyme is stimulated by divalent cations and carbohydrates but inhibited by sodium. Bioinformatic analysis indicates the enzyme is a membrane bound protein facing the extracellular side of the cell with 98% identity to the T. brucei homologous NTPDase gene.

Substrate specificity and enzyme recycling using chitosan immobilized laccase.

The immobilization of laccase (Aspergillus sp.) on chitosan by cross-linking and its application in bioconversion of phenolic compounds in batch reactors were studied. Investigation was performed using laccase immobilized via chemical cross-linking due to the higher enzymatic operational stability of this method as compared to immobilization via physical adsorption. To assess the influence of different substrate functional groups on the enzyme's catalytic efficiency, substrate specificity was investigated using chitosan-immobilized laccase and eighteen different phenol derivatives. It was observed that 4-nitrophenol was not oxidized, while 2,5-xylenol, 2,6-xylenol, 2,3,5-trimethylphenol, syringaldazine, 2,6-dimetoxyphenol and ethylphenol showed reaction yields up 90% at 40 °C. The kinetic of process, enzyme recyclability and operational stability were studied. In batch reactors, it was not possible to reuse the enzyme when it was applied to syringaldazne bioconversion. However, when the enzyme was applied to bioconversion of 2,6-DMP, the activity was stable for eight reaction batches.

Pedofilia: informações médico-legais para o profissional da saúde / Pedophilia: medical legal information for the health personnel

Todas as formas de violência contra a criança produzem consequências nefastas ao desenvolvimento infantil. Existe uma forte influência do trauma no progresso da arquitetura cerebral, da estruturação permanente da personalidade e dos padrões de relacionamento posteriores. Geralmente, os pedófilos iniciam suas atividades durante a adolescência, e a preferência pelo sexo masculino é duas vezes maior do que pelo feminino; sabe-se que as relações sexuais entre adultos e crianças ocorrem desde a antiguidade. Existem várias teorias para explicar a pedofilia: para alguns, a testosterona predispõe aos desvios de comportamento sexual; outros a explicam como resultado de fatores psicossociais. O diagnóstico é estabelecido preenchendo-se os critérios do Manual Diagnóstico e Estatístico de Transtornos Mentais (DSM-IV-TR). O tratamento de escolha é a psicoterapia de longa duração, e o tratamento medicamentoso se baseia em três classes de drogas: hormônios femininos, agonistas do LHRH ou antiandrogênios. A castração cirúrgica é oferecida em casos especiais. A prisão remove os pedófilos da sociedade, mas não os modifica. Além dos dispositivos constitucionais que preveem, de forma ampla e integral, a proteção da criança e do adolescente, o Código Penal e o Estatuto da Criança e do Adolescente estabelecem um sistema de normas imperativas destinadas à repressão de abusos envolvendo a temática em comento. Frente a essas novas revelações, faz-se mister ao profissional da saúde receber noções médico-legais sobre a violência e suas aberrações.(AU)

All forms of violence against children produce disastrous consequences to the infantile development. There is a strong influence of the trauma in the progress of the cerebral architecture, the permanent structure of the personality and the standard of posterior relationships. Pedophiles usually begin their activities during adolescence, and the preference for males is twice as high as for females. It is known that sexual relations between adults and children have occurred since ancient times. There is a handful of theories to explain pedophilia: for some, testosterone predisposes the individual to sexually deviant behaviors; for others, it is the result of psychosocial factors. The diagnosis is established by comparison with the criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR), and the treatment of choice is long-term psychotherapy; drug therapy is based on three classes of drugs: female hormones, LHRH agonists or anti-androgens. Surgical castration is resorted in special cases only. Imprisonment removes pedophiles from society, but does not change or cure them. Along with the constitutional provisions that offer, in a broad and integral manner, the protection of children and adolescents, the Criminal Code and the Statute of the Child and Adolescent establish a system of mandatory regulations for the prosecution of those abusive individuals concerning the matter herein discussed. Due to these new revelations, it is important that health personnel receive medical legal notions about the violence and its aberrations.(AU)

O profissional de saúde e a violência na infância e adolescência / The health professional and the violence in childhood and adolescence

A violência na infância e na adolescência é um grave fenômeno social e de saúde e não pode continuar sendo vista como uma responsabilidade policial. O Estatuto da Criança e do Adolescente reserva aos profissionais de saúde a atuação nos diagnósticos de maus tratos e os procedimentos com a notificação. O não cumprimento dessa responsabilidade ocorre devido à falta de conhecimento da lei ou por não estarem convencidos de que devem exercer esse papel. É importante afirmar que a notificação não é um ato pessoal, mas uma obrigação legal. Diante de uma vítima, o médico precisa seguir algumas normas que, por vezes, geram conflitos entre os preceitos legais e éticos. Na categoria médica é consensual comunicar as autoridades, sempre que se tratarem de menores de 14 anos e alguns casos de adolescentes. A evidência de maus tratos constitui justa causa para a quebra do sigilo. O paciente deve ser informado da possibilidade de um exame pericial no Instituto Médico Legal, capaz de uma avaliação mais criteriosa. Além da violência estrutural devido às desigualdades sociais existentes em nosso meio, as crianças e adolescentes também são violentadas dentro dos seus lares. Todas as formas de violência podem causar danos ao desenvolvimento biopsicossocial em curto, médio e longo prazos

The violence in childhood and adolescence is a serious social and health problem and it must not continue to be seen as the police's responsibility. The Statute of the Child and Adolescent assigns to health care professionals the action regarding diagnoses of injuries and procedures towards the notification. The unfulfillment of this responsibility is due to the lack of knowledge of the law or because they are not convinced that they must have this role. It is important to state that the notification is not a personal act, but a legal obligation. Facing the victim, the doctor must follow certain rules that sometimes conflict between legal and ethical ideas. In the medical it is consensual to communicate the authorities when the person is less than 14 years old and in some cases of adolescents. The evidence of mistreatment constitutes legal cause for confidentiality breach. The patient must be informed of the possibility of going through a peer review, in the Institute of Legal Medicine, that is capable of a more careful evaluation. In addition to the structural violence because of the social inequalities in our country, children and adolescents are also damaged inside their houses. All forms of violence can cause damage to the biopsychosocial development in short, medium and long terms

Recién nacidos de madres adolescentes precoces y tardías: ¿hay alguna diferencia? / Newborn from early and late pregnant adolescent

Introducción: los embarazos en la adolescencia llevan a situaciones que pueden atentar contra la salud de la madre, así como la del hijo, constituyendo un problema que no debe ser considerado solamente en los términos de hoy sino también para el futuro. Hay una gran preocupación en relación a la posibilidad de que la madre adolescente presente malos resultados prenatales. Objetivos: analizar características epidemiológicas y aspectos clínicos relevantes del recién nacido de madres adolescentes y comparar las variables analizadas entre las gestantes adolescentes precoces y tardías. Pacientes y métodos: en un estudio transversal, analítico, se evaluaron 2058 registros, siendo 322 del grupo de las precoces y 1736 las tardías, siendo las dos de la maternidad Escuela Assis Chateaubriand de la Universidad Federal de Ceará durante el periodo de un año. Resultados: del total de partos, en el periodo de un año, 26 por ciento eran adolescentes. El índice de prematurez fue de 16,7 por ciento (20,2 por ciento x 16,1por ciento, p = 0,069). El score de apgar 7 en el primer minuto fue 15,1por ciento (19,9 por ciento x 14,2 por ciento, p = 0,008). Otras variables: score de apgar en el quinto minuto, adecuando el peso a determinada edad de gestación malformaciones y muerte neo-natal presentaron índices bajos y sin una diferencia estadística entre los grupos. Conclusiones: no hubo diferencia estadísticamente significativa entre los grupos de edad entre las características de sus recién nacidos, contrariando afirmaciones bibliográficas.

Introduction: Teenage pregnancies can be an important problem for the health of the mother and the child, not only today but also for the future. Adolescent mothers have the possibility of poorly pre-natal outcome. Objective: We review relevant epidemiological and clinical aspects of newborn from pregnant adolescent and compare the early and late pregnant adolescents. Patients and methods: In a transversal, analytic study, we evaluated 322 early pregnant adolescent and 1736 late pregnant adolescent from the Assis Chateaubriand Maternity Hospital of the Federal University of Ceará during one year. Results In one year, 26 percent of the pregnant were teenagers. The pre-maturity rate was 16.7 percent (20.2 percent vs 16.1percent, p = 0.069). The Apgar score of 7 in the first minute was 15.1 percent (19.9 percent x 14.2 percent, p = 0.008). Other variables: Apgar score in the fifth minute, the birth weight to a given gestational age, birth and neonatal death rate, showed no statistical difference between groups. Conclusions: There was no statistically significant difference between age groups between the characteristics of their newborns, contrary to literature statements.

Hemangioma de mama en niña de cinco meses y estudio del caso / Breast hemangioma in a child: a case report

Se presenta el caso de un niño con un hemangioma de mama y se revisa la bibliografía al respecto.

It will be presented a case of a child with a breast hemangioma and a paper review.

Gestação na adolescência precoce e tardia: há diferença nos riscos obstétricos? / Precocious and late pregnancy in adolescents: is there a difference comparing the obstetric risks?

OBJETIVOS: descrever aspectos da assistência e resultados obstétricos da gravidez em adolescentes atendidas em um centro de atendimento terciário do Ceará e comparar os resultados maternos e perinatais entre adolescentes precoces e tardias. MÉTODOS: em estudo transversal, analítico, avaliaram-se 2.058 casos, sendo 322 (15,6 por cento) de adolescentes precoces e 1.736 (84,4 por cento) tardias, atendidas no ano de 2000. Foram analisados as intercorrências clínicas no pré-natal, tipo de parto, indicações de cesárea, idade gestacional no parto, peso do recém-nascido ao nascimento, adequação do peso à idade gestacional, índices de Apgar no primeiro e quinto minuto de vida, presença de malformações e óbito neonatal. Utilizaram-se o teste exato de Fisher e o chi2 na comparação entre os dois grupos. Calculou-se também a razão de prevalência. RESULTADOS: do total de partos ocorridos no período, 25,9 por cento eram de adolescentes e a média de idade destas foi de 17,2 anos. Constatou-se que 88 por cento freqüentaram o pré-natal, sendo 60 por cento com número insuficiente de consultas. As intercorrências clínicas mais freqüentes foram a pré-eclâmpsia (14,7 por cento), a anemia (12,9 por cento) e a infecção do trato urinário (6,4 por cento), sem diferença de freqüência entre os grupos. Ocorreram 31,3 por cento de nascimentos por cesárea, sendo a pré-eclâmpsia a principal indicação nas duas faixas etárias (25 e 23 por cento, respectivamente). A freqüência de Apgar menor que 7 no primeiro minuto foi de 19,9 por cento no grupo das adolescentes precoces e 14,2 por cento entre as tardias (x²=6,96, p=0,008). Não houve diferença quanto à freqüência de prematuridade (20,2 vs 16,1 por cento), recém-nascido pequeno para idade gestacional (12,4 vs 10,4 por cento), baixos escores de Apgar de quinto minuto (5,3 vs 3,3 por cento), malformações congênitas (3,1 vs 2,7 por cento) e morte neonatal (1,6 vs 3,1 por cento). CONCLUSÕES: as gestantes adolescentes...

PURPOSE: to describe the obstetric outcomes in pregnant adolescents at a tertiary hospital and to compare the maternal and labor outcomes between precocious and late adolescents. METHODS: in a transversal analytical study, 2058 cases were evaluated, considering 322 (15.65 percent) from the precocious group and 1736 (84.35 percent) from the late group that delivered at the "Maternidade Escola Assis Chateaubriand/UFC" from January 1, 2000 to December 31, 2000. The clinical complications in the prenatal period, kind of delivery, indications for cesarean section, birth gestational age at birth, birth weight, comparison of birth weight and gestational age, Apgar score at the first and fifth minute, presence of malformations, and neonatal death were analyzed. The exact Fisher and the chi2 tests were used to compare both groups. The prevalence ratio was calculated. RESULTS: from of total of deliveries, 25.95 percent belonged to adolescents. The average age was 17.19 years. Prenatal visits were made by 88 percent of the patients, but 60 percent had an insufficient number of visits. The most frequent clinical situations were preeclampsia (14.72 percent), anemia (12.97 percent) and urinary tract infections (6.37 percent), with no statistical difference between the groups. Thirty-one and three percent of the births were by cesarean section, preeclampsia being the main indication in the two age groups (25 and 23 percent, respectively). The frequency of an Apgar score less than 7 at the first minute was 19,9 percent in the precocious adolescent group and 14,2 percent in the late adolescent group (x²=6,96, p=0.008). There was no statistical difference regarding prematurity rate (20.2 vs 16.1 percent), low-birth weight infants (12.4 vs 10.4 percent), low Apgar score at the fifth minute (5.3 vs 3.3 percent), congenital malformations (3.1 vs 2.7 percent), and neonatal death (5.3 vs 3.3 percent). CONCLUSIONS: the precocious and late pregnant adolescents...

A proposta da rede de serviços sentinela como estratégia da vigilância de violências e acidentes / The injury surveillance system based on sentinel health services

No Brasil, as bases de dados oficiais permitem o monitoramento da mortalidade e internações no SUS, decorrentes dos acidentes e violências. É preciso conhecer a magnitude e o perfil dessas causas que demandam os serviços de emergência, bem como identificar alguns problemas ocultos tais como as violências doméstica e sexual. O propósito deste artigo é apresentar a proposta do Ministério da Saúde de implantação da Rede de Serviços Sentinela de Vigilância de Violências e Acidentes - Rede VIVA, iniciada em 2006, que visa complementar o sistema de informações existente para a vigilância dessas causas. Para obter um quadro mais completo do problema e atender à legislação vigente no País, foram estabelecidos dois componentes: 1) Vigilância de acidentes e violências em emergências hospitalares selecionadas: coleta em um mês a cada ano, através de uma amostra; 2) Vigilância das violências sexual, doméstica e/ou outras violências interpessoais em serviços de referência: coleta universal e contínua. O estabelecimento da Rede VIVA foi realizado pelo Ministério da Saúde em parceria com as Secretarias Estaduais e Municipais de Saúde a partir de critérios previamente estabelecidos. A adesão ao projeto foi acima das expectativas, todas as regiões do Brasil foram representadas.

In Brazil, the official data sets allow monitoring the impact of injury deaths and injury hospitalization in the public health system. But it is necessary to gather more information about the magnitude and the characteristics of injuries at Emergency Departments (ED), as well as to identify some hidden problems, such as domestic and sexual violence. The purpose of this article is to present the new Injury Surveillance System based on Sentinel Health Services, carried out by the Ministry of Health in order to broaden the knowledge of these causes.To have a more accurate picture of injuries and to enforce the law which made mandatory the information about violence against women in the country, the measures to be taken were twofold: 1) injury surveillance in ED, carried out in chosen services, collecting one-month data yearly, through a sample; 2) domestic, sexual and interpersonal violence surveillance carried out in violence reference services, through universal and continuous data collection, involving a larger number of services. The implementation of that Health Sentinel Services Network has been conducted by the Ministry of Health in partnership with the State and Municipal Health Departments based on pre-established criteria. The adherence to the project has been taken place all over Brazil.