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1.
BMC Pediatr ; 22(1): 468, 2022 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-35922776

RESUMO

BACKGROUND: Infantile colic is a common problem during the first three months of life. This randomized, double-blind, placebo-controlled trial conducted in an urban hospital in Delhi, India evaluated the efficacy and safety of oral lactase in management of infantile colic. METHODS: One hundred sixty-two clinically healthy infants aged < 5 months age [mean (SD) = 63.5 (30.5) days] fulfilling the Rome-IV diagnostic criteria for infantile colic were enrolled. Eligible children were randomly allocated to receive 5 drops of lactase (600 FCC units/mL) (n = 80) or placebo (n = 82) mixed with breast milk or formula feed four times a day for a duration of 4 weeks. Primary outcomes were duration of crying or fussing (min/d), and number of days with colic lasting > 3 h/d; secondary outcomes were parental satisfaction and adverse events. RESULTS: At the end of four weeks, mean (SD) crying or fussing time (min/d) was significantly shorter in infants receiving lactase in comparison to placebo [89.9 (115.2) vs.178.5 (153.2); P = 0.001]. The mean (SD) number of days with colic was also significantly less in the lactase group as compared to placebo group at the end of the treatment [12.1 (7.8) vs 17.6 (8.4); P < 0.001]. By the end of 4th week, parental satisfaction in terms of infant's mood, activity, alertness, comfort and oral intake was better in intervention group. The adverse event profile was comparable between two groups. CONCLUSIONS: Oral lactase treatment in infantile colic results in symptomatic relief in terms of shortening of duration of crying or fussing, and better parental satisfaction. TRIAL REGISTRATION: Clinical trial registry of India (CTRI/2017/12/010930) registered on 20/12/2017.


Assuntos
Cólica , Criança , Cólica/tratamento farmacológico , Choro , Método Duplo-Cego , Feminino , Humanos , Lactente , Lactase/uso terapêutico , Pais
2.
Int J Mol Sci ; 23(14)2022 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-35887087

RESUMO

PLG-007 is a developmental therapeutic compound that has been clinically shown to reduce the magnitude of postprandial glucose excursions and has the potential to be an adjunct treatment for diabetes and inflammatory-related diseases. The present investigation is aimed at understanding the molecular mechanism of action of PLG-007 and its galactomannan (GM) components GMα and GMß (in a 1:4 mass ratio, respectively) on enzyme (i.e., α-amylase, maltase, and lactase) hydrolysis of glucose polymers using colorimetric assays and 13C HSQC NMR spectroscopy. The starch-iodine colorimetric assay indicated that GMα strongly inhibits α-amylase activity (~16-fold more potent than GMß) and thus is the primary active component in PLG-007. 13C HSQC experiments, used to follow the α-amylase-mediated hydrolysis of starch and amylopectin, further demonstrate the α-amylase inhibitory effect of GMα via α-amylase-mediated hydrolysis of starch and amylopectin. Maltohexaose (MT6) was used to circumvent the relative kinetic complexity of starch/amylopectin degradation in Michaelis-Menten analyses. The Vmax, KM, and Ki parameters were determined using peak volume integrals from 13C HSQC NMR spectra. In the presence of PLG-007 with α-amylase and MT6, the increase in KM from 7.5 ± 0.6 × 10-3 M (control) to 21 ± 1.4 × 10-3 M, with no significant change in Vmax, indicates that PLG-007 is a competitive inhibitor of α-amylase. Using KM values, Ki was estimated to be 2.1 ± 0.9 × 10-6 M; however, the microscopic Ki value of GMα is expected to be larger as the binding stoichiometry is likely to be greater than 1:1. Colorimetric assays also demonstrated that GMα is a competitive inhibitor of the enzymes maltase and lactase. Overall, this study provides insight as to how PLG-007 (GMα) is likely to function in vivo.


Assuntos
Amilopectina , alfa-Glucosidases , Amilopectina/química , Galactose/análogos & derivados , Glucose , Hidrólise , Lactase , Mananas , Amido/metabolismo , alfa-Amilases/metabolismo , alfa-Glucosidases/metabolismo
3.
Science ; 377(6605): 456, 2022 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-35901158

RESUMO

Giant study of ancient pottery and DNA challenges common evolutionary explanation for lactase persistence.


Assuntos
Lactase , Leite , Animais , DNA , Fazendas , Humanos , Lactase/genética , Brancos
4.
Curr Opin Clin Nutr Metab Care ; 25(5): 334-340, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-35838278

RESUMO

PURPOSE OF REVIEW: To provide an up-to-date review on the clinical assessment of two important gastrointestinal problems with overlapping symptomatology but diverse cause and testing methods. Small intestinal bacterial overgrowth (SIBO) is characterized by the presence of excess bacteria in the small intestine associated with bloating, distention, gas, and diarrhea. Lactose intolerance is caused by lactase enzyme deficiency in the small bowel mucosa leading to lactose malabsorption and symptoms of bloating, gas, and diarrhea. RECENT FINDINGS: SIBO is assessed by hydrogen/methane breath test using glucose as a substrate and/or small bowel aspirate and culture but these tests have shortcomings. Consequently, several new diagnostic techniques, including novel capsule technologies and other approaches are being evaluated. Lactose intolerance can be assessed by hydrogen/methane breath test using lactose as a substrate, or small bowel mucosal lactase assay, genetic testing and lactose tolerance test, although the efficacy and practicality of these diagnostic modalities are not equal. SUMMARY: In clinical practice, gas, bloating, distention, pain, and diarrhea are common gastrointestinal symptoms that often remain unexplained when routine gastrointestinal endoscopy, imaging, and stool tests are negative. These patients should be evaluated for SIBO and/or food intolerances including lactose intolerance.


Assuntos
Intolerância à Lactose , Testes Respiratórios/métodos , Diarreia/diagnóstico , Diarreia/etiologia , Humanos , Hidrogênio , Lactase , Intolerância à Lactose/diagnóstico , Intolerância à Lactose/microbiologia , Metano
5.
Food Res Int ; 157: 111004, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35761512

RESUMO

Lactose intolerance in infants can be overcome by adding lactase enzyme formulations to milk, hydrolysing the lactose present. Commonly, such formulations require a significant incubation period after addition to milk to allow time for hydrolysis to take place at desired temperature. Such incubation is often problematic for household conditions, and therefore, formulations with short or, ideally, no incubation period are needed. Development of such formulations requires understanding of the complex process of lactose hydrolysis within the variable temperature and pH conditions in the milk bottle and in the gastrointestinal tract. The current paper describes the application of high-resolution ultrasonic spectroscopy for detailed characterisation of the effect of pH, temperature, and proteases present in the digestive tract on hydrolysis of lactose. The results were employed in the development of a predictive model of lactose hydrolysis within conditions of the infant digestive system. The model was applied for analysis of the whole infant feeding process: from hydrolysis in the bottle, to the stomach and small intestine, and for assessment of the impact of enzyme dosage, the incubation period, and bottle temperature profile on the levels of lactose and galacto-oligosaccharides (produced during hydrolysis), released from the stomach and reaching the large intestine where symptoms of intolerance arise. The results showed that the activity of Ha-Lactase in the stomach and small intestine assists significantly in the reduction of lactose load to 10 % or less of the amount of lactose in the feed, depending on conditions.


Assuntos
Lactase , Lactose , Animais , Sistema Digestório , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Lactente , Lactose/química , Leite/química , Peptídeo Hidrolases , Temperatura , Ultrassom
6.
Nat Genet ; 54(2): 134-142, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35115689

RESUMO

Human genetic variation affects the gut microbiota through a complex combination of environmental and host factors. Here we characterize genetic variations associated with microbial abundances in a single large-scale population-based cohort of 5,959 genotyped individuals with matched gut microbial metagenomes, and dietary and health records (prevalent and follow-up). We identified 567 independent SNP-taxon associations. Variants at the LCT locus associated with Bifidobacterium and other taxa, but they differed according to dairy intake. Furthermore, levels of Faecalicatena lactaris associated with ABO, and suggested preferential utilization of secreted blood antigens as energy source in the gut. Enterococcus faecalis levels associated with variants in the MED13L locus, which has been linked to colorectal cancer. Mendelian randomization analysis indicated a potential causal effect of Morganella on major depressive disorder, consistent with observational incident disease analysis. Overall, we identify and characterize the intricate nature of host-microbiota interactions and their association with disease.


Assuntos
Dieta , Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Variação Genética , Interações entre Hospedeiro e Microrganismos , Polimorfismo de Nucleotídeo Único , Sistema ABO de Grupos Sanguíneos/genética , Bifidobacterium/fisiologia , Clostridiales/fisiologia , Estudos de Coortes , Neoplasias Colorretais/genética , Neoplasias Colorretais/microbiologia , Transtorno Depressivo Maior/genética , Transtorno Depressivo Maior/microbiologia , Fibras na Dieta , Enterococcus faecalis/fisiologia , Microbioma Gastrointestinal/genética , Estudo de Associação Genômica Ampla , Humanos , Lactase/genética , Complexo Mediador/genética , Análise da Randomização Mendeliana , Metagenoma , Morganella/fisiologia
7.
Am J Gastroenterol ; 117(5): 721-728, 2022 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-35169106

RESUMO

Many clinicians have suboptimal knowledge of evolutionary medicine. This discipline integrates social and basic sciences, epidemiology, and clinical medicine, providing explanations, especially ultimate causes, for many conditions. Principles include genetic variation from population bottleneck and founder effects, evolutionary trade-offs, and coevolution. For example, host-microbe coevolution contributes to the inflammatory and carcinogenic variability of Helicobacter pylori. Antibiotic-resistant strains are evolving, but future therapy could target promutagenic proteins. Ancient humans practicing dairying achieved survival and reproduction advantages of postweaning lactase persistence and passed this trait to modern descendants, delegitimizing lactose intolerance as "disease" in people with lactase nonpersistence. Three evolutionary hypotheses are each relevant to multiple diseases: (i) the polyvagal hypothesis posits that prehistoric adaptation of autonomic nervous system reactions to stress is beneficial acutely but, when continued chronically, predisposes individuals to painful functional gastrointestinal disorders, in whom it may be a biomarker; (ii) the thrifty gene hypothesis proposes genetic adaptation to feast-famine cycles among Pleistocene migrants to America, which is mismatched with Indigenous Americans' current diet and physical activity, predisposing them to obesity, nonalcoholic fatty liver disease, and gallstones and their complications; and (iii) the hygiene hypothesis proposes alteration of the gut microbiome, with which humans have coevolved, in allergic and autoimmune disease pathogenesis; for example, association of microbiome-altering proton pump inhibitor use with pediatric eosinophilic esophagitis, early-life gastrointestinal infection with celiac disease, and infant antibiotic use and an economically advanced environment with inflammatory bowel disease. Evolutionary perspectives broaden physicians' understanding of disease processes, improve care, and stimulate research.


Assuntos
Doenças do Sistema Digestório , Gastroenteropatias , Helicobacter pylori , Intolerância à Lactose , Antibacterianos , Evolução Biológica , Farmacorresistência Bacteriana , Humanos , Lactase/genética , Lactose/metabolismo , Intolerância à Lactose/genética
8.
Nat Genet ; 54(2): 100-106, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35115688

RESUMO

The human gut microbiome is a complex ecosystem that is involved in its host's metabolism, immunity and health. Although interindividual variations in gut microbial composition are mainly driven by environmental factors, some gut microorganisms are heritable and thus can be influenced by host genetics. In the past 5 years, 12 microbial genome-wide association studies (mbGWAS) with >1,000 participants have been published, yet only a few genetic loci have been consistently confirmed across multiple studies. Here we discuss the state of the art for mbGWAS, focusing on current challenges such as the heterogeneity of microbiome measurements and power issues, and we elaborate on potential future directions for genetic analysis of the microbiome.


Assuntos
Microbioma Gastrointestinal , Variação Genética , Estudo de Associação Genômica Ampla , Locos de Características Quantitativas , Sistema ABO de Grupos Sanguíneos/genética , Variação Biológica da População , Fucosiltransferases/genética , Microbioma Gastrointestinal/genética , Genômica , Interações entre Hospedeiro e Microrganismos , Humanos , Lactase/genética , Polimorfismo de Nucleotídeo Único
9.
Nat Genet ; 54(2): 143-151, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35115690

RESUMO

Host genetics are known to influence the gut microbiome, yet their role remains poorly understood. To robustly characterize these effects, we performed a genome-wide association study of 207 taxa and 205 pathways representing microbial composition and function in 7,738 participants of the Dutch Microbiome Project. Two robust, study-wide significant (P < 1.89 × 10-10) signals near the LCT and ABO genes were found to be associated with multiple microbial taxa and pathways and were replicated in two independent cohorts. The LCT locus associations seemed modulated by lactose intake, whereas those at ABO could be explained by participant secretor status determined by their FUT2 genotype. Twenty-two other loci showed suggestive evidence (P < 5 × 10-8) of association with microbial taxa and pathways. At a more lenient threshold, the number of loci we identified strongly correlated with trait heritability, suggesting that much larger sample sizes are needed to elucidate the remaining effects of host genetics on the gut microbiome.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Fenômenos Fisiológicos Bacterianos , Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Variação Genética , Interações entre Hospedeiro e Microrganismos , Lactase/genética , Bifidobacterium/fisiologia , Dieta , Fucosiltransferases/genética , Genoma Humano , Estudo de Associação Genômica Ampla , Humanos , Redes e Vias Metabólicas , Metagenoma , Herança Multifatorial , Países Baixos , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Cloreto de Sódio na Dieta , Triglicerídeos/sangue
10.
Arch. argent. pediatr ; 120(1): 59-66, feb 2022. tab, ilus
Artigo em Inglês, Espanhol | LILACS, BINACIS | ID: biblio-1353500

RESUMO

La lactosa es el principal carbohidrato de la leche materna. Es un disacárido conformado por glucosa y galactosa. Su producción en la glándula mamaria es independiente de la dieta materna. Además de proveer energía, es la única fuente de galactosa de la dieta, necesaria para la síntesis de macromoléculas como oligosacáridos, glicoproteínas y glicolípidos. Favorece la absorción y retención de calcio, magnesio y cinc. Su digestión por la enzima lactasa y posterior absorción tienen lugar en intestino delgado. El déficit de lactasa, que puede ser primario congénito (muy infrecuente), primario tardío o secundario por lesión intestinal, puede generar intolerancia con síntomas como dolor, distensión abdominal, flatulencia y diarrea. En el colon, bifidobacterias y lactobacilos pueden hidrolizarla. El manejo nutricional de la intolerancia deberá hacerse siempre preservando la lactancia materna. La reducción o suspensión de la lactosa deberá ser transitoria y se reemplazarán alimentos suspendidos por otros con adecuados aportes calóricos, proteicos y de minerales y vitaminas.


Lactose is the main carbohydrate present in humanmilk. It is a disaccharide made up of glucoseand galactose. It is produced in the mammaryglands, regardless of maternal diet. In addition toproviding energy, it is the only source of dietarygalactose, necessary for macromolecule synthesis,including oligosaccharides, glycoproteins, andglycolipids. It favors calcium, magnesium, andzinc absorption and retention. Its digestion bylactase and subsequent absorption occurs inthe small intestine. Lactase deficiency may beclassified into congenital primary (very rare),late-onset primary or secondary due to an injuryof the intestine; it may cause intolerance withpain, abdominal distension, abdominal gas, anddiarrhea. In the colon, it may be hydrolyzed bybifidobacteria and lactobacilli. The nutritionalmanagement of intolerance should alwayspreserve breastfeeding. Lactose reduction orelimination should be transient, and eliminatedfood should be replaced with other similar incalorie, protein, mineral, and vitamin content.


Assuntos
Humanos , Intolerância à Lactose/diagnóstico , Lactase/metabolismo , Dieta , Lactose/metabolismo , Leite Humano/metabolismo
11.
Biochim Biophys Acta Mol Basis Dis ; 1868(4): 166338, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35007711

RESUMO

Patients with the rare autosomal recessive disorder congenital lactase deficiency (CLD) present with severe, potentially life-threatening symptoms shortly after birth. Several variants have been characterized within the gene for lactase-phlorizin hydrolase (LCT) that are associated with CLD. Here, we analyze at the biochemical and cellular levels LCT mutants harboring the genetic variants p.Y1390*, p.E1612*, p.S1150Pfs*19, p.S1121L, p.R1587H, and p.S688P. Our data unequivocally demonstrate that these mutants are absolutely transport incompetent, some of which are readily degraded, and are enzymatically inactive. The current study contributes to and expands our understanding on the pathogenesis of CLD at the molecular level.


Assuntos
Erros Inatos do Metabolismo dos Carboidratos/patologia , Lactase-Florizina Hidrolase/genética , Lactase/deficiência , Animais , Células COS , Erros Inatos do Metabolismo dos Carboidratos/genética , Chlorocebus aethiops , Humanos , Lactase/genética , Lactase-Florizina Hidrolase/química , Lactase-Florizina Hidrolase/metabolismo , Mutagênese Sítio-Dirigida , Mutação de Sentido Incorreto , Dobramento de Proteína , Transporte Proteico
12.
Arch Argent Pediatr ; 120(1): 59-66, 2022 02.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-35068123

RESUMO

Lactose is the main carbohydrate present in human milk. It is a disaccharide made up of glucose and galactose. It is produced in the mammary glands, regardless of maternal diet. In addition to providing energy, it is the only source of dietary galactose, necessary for macromolecule synthesis, including oligosaccharides, glycoproteins, and glycolipids. It favors calcium, magnesium, and zinc absorption and retention. Its digestion by lactase and subsequent absorption occurs in the small intestine. Lactase deficiency may be classified into congenital primary (very rare), late-onset primary or secondary due to an injury of the intestine; it may cause intolerance with pain, abdominal distension, abdominal gas, and diarrhea. In the colon, it may be hydrolyzed by bifidobacteria and lactobacilli. The nutritional management of intolerance should always preserve breastfeeding. Lactose reduction or elimination should be transient, and eliminated food should be replaced with other similar in calorie, protein, mineral, and vitamin content.


La lactosa es el principal carbohidrato de la leche materna. Es un disacárido conformado por glucosa y galactosa. Su producción en la glándula mamaria es independiente de la dieta materna. Además de proveer energía, es la única fuente de galactosa de la dieta, necesaria para la síntesis de macromoléculas como oligosacáridos, glicoproteínas y glicolípidos. Favorece la absorción y retención de calcio, magnesio y cinc. Su digestión por la enzima lactasa y posterior absorción tienen lugar en intestino delgado. El déficit de lactasa, que puede ser primario congénito (muy infrecuente), primario tardío o secundario por lesión intestinal, puede generar intolerancia con síntomas como dolor, distensión abdominal, flatulencia y diarrea. En el colon, bifidobacterias y lactobacilos pueden hidrolizarla. El manejo nutricional de la intolerancia deberá hacerse siempre preservando la lactancia materna. La reducción o suspensión de la lactosa deberá ser transitoria y se reemplazarán alimentos suspendidos por otros con adecuados aportes calóricos, proteicos y de minerales y vitaminas.


Assuntos
Intolerância à Lactose , Dieta , Humanos , Lactase/metabolismo , Lactose/metabolismo , Intolerância à Lactose/diagnóstico , Leite Humano/metabolismo
13.
Eur J Epidemiol ; 37(7): 713-722, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34978666

RESUMO

BACKGROUND: Previous observational studies have indicated a protective effect of drinking milk on asthma and allergy. In Mendelian Randomization, one or more genetic variants are used as unbiased markers of exposure to examine causal effects. We examined the causal effect of milk intake on hay fever, asthma, forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) by using the lactase rs4988235 genotype associated with milk intake. METHODS: We performed a Mendelian Randomization study including 363,961 participants from the UK Biobank. RESULTS: Observational analyses showed that self-reported milk-drinkers vs. non-milk drinkers had an increased risk of hay fever: odds ratio (OR) = 1.36 (95% CI 1.32, 1.40, p < 0.001), asthma: OR = 1.33 (95% CI 1.38, 1.29, p < 0.001), yet a higher FEV1: ß = 0.022 (SE = 0.004, p < 0.001) and FVC: ß = 0.026 (SE = 0.005, p < 0.001). In contrast, genetically determined milk-drinking vs. not drinking milk was associated with a lower risk of hay fever: OR = 0.791 (95% CI 0.636, 0.982, p = 0.033), and asthma: OR = 0.587 (95% CI 0.442, 0.779, p = 0.001), and lower FEV1: ß = - 0.154 (standard error, SE = 0.034, p < 0.001) liter, and FVC: ß = - 0.223 (SE = 0.034, p < 0.001) liter in univariable MR analyses. These results were supported by multivariable Mendelian randomization analyses although not statistically significant. CONCLUSIONS: As opposed to observational results, genetic association findings indicate that drinking milk has a protective effect on hay fever and asthma but may also have a negative effect on lung function. The results should be confirmed in other studies before any recommendations can be made.


Assuntos
Asma , Rinite Alérgica Sazonal , Asma/epidemiologia , Asma/genética , Humanos , Lactase/genética , Pulmão , Análise da Randomização Mendeliana , Rinite Alérgica Sazonal/genética
14.
Ann Hum Genet ; 86(1): 24-33, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34523124

RESUMO

Although imputation of missing SNP results has been widely used in genetic studies, claims about the quality and usefulness of imputation have outnumbered the few studies that have questioned its limitations. But it is becoming clear that these limitations are real-for example, disease association signals can be missed in regions of LD breakdown. Here, as a case study, using the chromosomal region of the well-known lactase gene, LCT, we address the issue of imputation in the context of variants that have become frequent in a limited number of modern population groups only recently, due to selection. We study SNPs in a 500 bp region covering the enhancer of LCT, and compare imputed genotypes with directly genotyped data. We examine the haplotype pairs of all individuals with discrepant and missing genotypes. We highlight the nonrandom nature of the allelic errors and show that most incorrect imputations and missing data result from long haplotypes that are evolutionarily closely related to those carrying the derived alleles, while some relate to rare and recombinant haplotypes. We conclude that bias of incorrectly imputed and missing genotypes can decrease the accuracy of imputed results substantially.


Assuntos
Lactase , Polimorfismo de Nucleotídeo Único , Alelos , Frequência do Gene , Genótipo , Haplótipos , Humanos , Lactase/genética
15.
Anal Biochem ; 631: 114364, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34487718

RESUMO

Ricin is a toxic protein derived from the castor bean plant (Ricinus communis) and has potential for bioterrorism or criminal use. Therefore, sensitive and rapid analytical methods are needed for its confirmatory detection in environmental samples. Our laboratory previously reported on the development of a confirmatory method to detect ricin involving antibody capture of ricin followed by mass spectrometric detection of ricin's enzymatic activity and of tryptic fragments unique to ricin. Here, we describe a novel ricin capture method of magnetic beads coated with 4-aminophenyl-1-thiol-ß-galactopyranoside, using ricin's lectin characteristics. The assay has been adapted for use on a simple, benchtop MALDI-TOF MS mass spectrometer common in clinical microbiology laboratories. Validation of the novel assay includes establishment of a limit of detection, and an examination of assay selectivity. The limit of detection of the enzymatic activity method is 8 ng/mL and 500 ng/mL for the confirmatory tryptic fragment assay. The assay is highly selective with no cross-reactivity from near neighbors and highly specific with a panel of 19 cultivars all testing positive. Additionally, there were no interferences found during testing of a panel of white powders. This allows for a confirmatory detection method for ricin in laboratories lacking expensive, sophisticated mass spectrometers.


Assuntos
Microesferas , Ricina/análise , Ricina/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Anticorpos/química , Contaminação de Alimentos/análise , Galactose/química , Lactase/química , Limite de Detecção , Fenômenos Magnéticos , Leite/química , Extratos Vegetais/análise , Pós/análise , Pós/química , Reprodutibilidade dos Testes , Ricina/metabolismo , Ricinus/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação , Tripsina/química
16.
Mol Biol Rep ; 48(11): 7087-7093, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34515921

RESUMO

BACKGROUND: In adulthood the activity of the lactase enzyme is inherited as autosomal dominant form associated to Single nucleotide polymorphisms (SNPs). The present research was aimed to develop a novel genetic method to test lactase non persistence more powerfully. METHODS AND RESULTS: In our study, we selected eight different SNPs that are associated with lactase persistence from Caucasian, Arabian Bedouins, sub-Saharian Africans and Asian populations to set up an approach to detect all the eight different SNPs at the same time in the same sample. This technique is centred on the identification of SNPs with a single nucleotide primer extension method using Sanger sequencing and capillary electrophoresis. CONCLUSIONS: Our method allowed us to check the genotype asset of eight SNPs related to lactase persistence simultaneously and in a very efficient manner. It could be applied to a higher number of SNPs in a single reaction.


Assuntos
Lactase/deficiência , Intolerância à Lactose , Polimorfismo de Nucleotídeo Único , Adulto , Feminino , Humanos , Lactase/química , Lactase/genética , Lactase/metabolismo , Intolerância à Lactose/enzimologia , Intolerância à Lactose/genética , Masculino , Pessoa de Meia-Idade
17.
J Dairy Res ; 88(3): 357-365, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34425920

RESUMO

The aim of this review was to present various topics related to lactose intolerance with special attention given to the role of fermented foods and probiotics in alleviating gastrointestinal symptoms. Lactose intolerance is a common digestive problem in which the human body is unable to digest lactose, known as milk sugar. Lactose intolerance can either be hereditary or a consequence of intestinal diseases. Recent work has demonstrated that fermented dairy products and probiotics can modify the metabolic activities of colonic microbiota and may alleviate the symptoms of lactose intolerance. We suggest that, lactose free dairy products could be recommended as alternatives for the alleviation of lactose intolerance and for the promotion of human health and wellness.


Assuntos
Intolerância à Lactose/terapia , Probióticos , Animais , Colo/microbiologia , Laticínios/análise , Microbioma Gastrointestinal/fisiologia , Humanos , Lactase/deficiência , Lactase/metabolismo , Lactose/análise , Lactose/metabolismo , Probióticos/uso terapêutico
18.
Int J Biol Macromol ; 189: 410-419, 2021 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-34437917

RESUMO

We have previously demonstrated the ability of the human vaginal strain Lactobacillus crispatus 2029 (LC2029) for strong adhesion to cervicovaginal epithelial cells, expression of the surface layer protein 2 (Slp2), and antagonistic activity against urogenital pathogens. Slp2 forms regular two-dimensional structure around the LC2029 cells,which is secreted into the medium and inhibits intestinal pathogen-induced activation of caspase-9 and caspase-3 in the human intestinal Caco-2 cells. Here, we elucidated the effects of soluble Slp2 on adhesion of proteobacteria pathogens inducing necrotizing enterocolitis (NEC), such as Escherichia coli ATCC E 2348/69, E. coli ATCC 31705, Salmonella Enteritidis ATCC 13076, Campylobacter jejuni ATCC 29428, and Pseudomonas aeruginosa ATCC 27853 to Caco-2 cells, as well as on growth promotion, differentiation, vascular endothelial growth factor (VEGF) production, and intestinal barrier function of Caco-2 cell monolayers. Slp2 acts as anti-adhesion agent for NEC-inducing proteobacteria, promotes growth of immature Caco-2 cells and their differentiation, and enhances expression and functional activity of sucrase, lactase, and alkaline phosphatase. Slp2 stimulates VEGF production, decreases paracellular permeability, and increases transepithelial electrical resistance, strengthening barrier function of Caco-2 cell monolayers. These data support the important role of Slp2 in the early postnatal development of the human small intestine enterocytes.


Assuntos
Diferenciação Celular , Enterócitos/metabolismo , Lactobacillus crispatus/química , Glicoproteínas de Membrana/farmacologia , Vagina/microbiologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Aderência Bacteriana/efeitos dos fármacos , Células CACO-2 , Diferenciação Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Impedância Elétrica , Enterócitos/efeitos dos fármacos , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Lactase/genética , Lactase/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sacarase/genética , Sacarase/metabolismo
19.
Nutrients ; 13(8)2021 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-34445060

RESUMO

Milk intake has been associated with risk of neurodegenerative diseases in observational studies. Nevertheless, whether the association is causal remains unknown. We adopted Mendelian randomization design to evaluate the potential causal association between milk intake and common neurodegenerative diseases, including multiple sclerosis (MS), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), and Parkinson's disease (PD). Genetic associations for neurodegenerative diseases were obtained from the International Multiple Sclerosis Genetics Consortium (n = 80,094), FinnGen consortium (n = 176,899), AD GWAS (n = 63,926), Web-Based Study of Parkinson's Disease (n = 308,518), PDGene (n = 108,990), and ALS GWAS (n = 80,610). Lactase persistence variant rs4988235 (LCT-13910 C > T) was used as the instrumental variable for milk intake. Genetically predicted higher milk intake was associated with a decreased risk of MS and AD and with an increased risk of PD. For each additional milk intake increasing allele, the odds ratios were 0.94 (95% confidence intervals [CI]: 0.91-0.97; p = 1.51 × 10-4) for MS, 0.97 (0.94-0.99; p = 0.019) for AD and 1.09 (95%CI: 1.06-1.12, p = 9.30 × 10-9) for PD. Genetically predicted milk intake was not associated with ALS (odds ratio: 0.97, 95%CI: 0.94-1.01, p = 0.135). Our results suggest that genetically predicted milk intake is associated with a decreased risk of MS and AD but with an increased risk of PD. Further investigations are needed to clarify the underlying mechanisms.


Assuntos
Interação Gene-Ambiente , Lactase/genética , Leite , Doenças Neurodegenerativas/genética , Polimorfismo de Nucleotídeo Único , Doença de Alzheimer/epidemiologia , Doença de Alzheimer/genética , Esclerose Amiotrófica Lateral , Animais , Estudos de Casos e Controles , Bases de Dados Genéticas , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Análise da Randomização Mendeliana , Leite/efeitos adversos , Esclerose Múltipla/epidemiologia , Esclerose Múltipla/genética , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/epidemiologia , Doença de Parkinson/epidemiologia , Doença de Parkinson/genética , Medição de Risco , Fatores de Risco
20.
Biochem Mol Biol Educ ; 49(6): 935-941, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34406692

RESUMO

A laboratory exercise for undergraduate advanced students of enzymology and biocatalysis is presented. Since enzyme assays can be quenched or continuous, this experiment compares the performance of two quenching agents for lactase, in a continuous setup. Enzymatic activity of ß-galactosidase (Aspergillus oryzae) was determined based on the release of 4-nitrophenol from 4-nitrophenyl ß-D-galactopyranoside using a microplate reader. Sodium carbonate and borate buffer were tested as quenching agents, and experimental control was the unstopped assay. Based on released 4-nitrophenol, enzyme activity, and rate constant k, the students could assess the performance of each termination agent. The experiment promotes disciplinary and transversal competencies, including research-based learning, critical thinking, and introduce the students to high-throughput techniques that are common in the research and development environment.


Assuntos
Boratos , Lactase , Carbonatos , Humanos , Laboratórios , beta-Galactosidase
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