RESUMO
Criminal traces commonly found at crime scenes may present mixtures from two or more individuals. The scene of the crime is important for the collection of various types of traces in order to find the perpetrator of the crime. Thus, we propose that hematophagous mosquitoes found at crime scenes can be used to perform genetic testing of human blood and aid in suspect investigation. The aim of the study was to obtain a single Aedes aegypti mosquito profile from a human DNA mixture containing genetic materials of four individuals. We also determined the effect of blood acquisition time by setting time intervals of 24, 48, and 72 h after the blood meal. STR loci and amelogenin were analyzed, and the results showed that human DNA profiles could be obtained from hematophagous mosquitos at 24 h following the blood meal. It is possible that hematophagous mosquitoes can be used as biological remains at the scene of the crime, and can be used to detect human DNA profiles of up to four individuals.
Assuntos
Aedes/química , Impressões Digitais de DNA/métodos , DNA/isolamento & purificação , Genética Forense/métodos , Aedes/fisiologia , Animais , Mordeduras e Picadas/sangue , Células Sanguíneas/química , Crime , DNA/genética , Feminino , Testes Genéticos/métodos , Voluntários Saudáveis , Humanos , Masculino , Repetições de MicrossatélitesRESUMO
Investigations of any type of crime invariably starts at the crime scene by collecting evidence. Thus, the purpose of this research was to collect and analyze an entomological trace from an environment that is similar to those of indoor crime scenes. Hematophagous mosquitoes were collected from two residential units; saliva of volunteers that were residents in the units was also collected for genetic analysis as reference samples. We examined the allele frequencies of 15 short tandem repeat loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) and amelogenin. A total of 26 female hematophagous mosquitoes were identified as Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus; we were able to obtain 11 forensically valid genetic profiles, with a minimum of 0.028203 ng/µL of human DNA. Thus, the results of this study showed that it was possible to correlate human genetic information from mosquitoes with the volunteer reference samples, which validates the use of this information as forensic evidence. Furthermore, we observed mixed genetic profiles from one mosquito. Therefore, it is clearly important to collect these insects indoors where crimes were committed, because it may be possible to find intact genetic profiles of suspects in the blood found in the digestive tract of hematophagous mosquitoes for later comparison to identify an offender and/or exclude suspects.
Assuntos
Culicidae/genética , DNA/sangue , Genética Forense , Repetições de Microssatélites/genética , Animais , Crime , Impressões Digitais de DNA , Feminino , Frequência do Gene , Testes Genéticos , HumanosRESUMO
Este trabalho teve como objetivo avaliar a composição química e as características fermentativas de silagens da polpa de café com diferentes proporções de casca de café. O material foi ensilado em PVC de 150 mm de diâmetro por 750 mm de altura, segundo os tratamentos: polpa de café (PC), PC + 20% de casca de café (CC), PC + 40% de CC e PC + 60% de CC, segundo delineamento inteiramente casualizado com seis repetições. Os silos foram abertos 60 dias após o fechamento, quando foram retiradas amostras para determinação dos teores de matéria seca (MS), proteína bruta (PB), fibra em detergente neutro (FDN), fibra em detergente ácido (FDA), lignina, celulose, pH, N-NH3, cafeína, cálcio (Ca), fósforo (P) e digestibilidade in vitro da MS (DIVMS). Determinou-se também a produção de efluentes e de gás. Houve um aumento linear no teor de MS e pH, à medida que foi adicionado casca de café nas silagens, e decréscimo linear nos teores de PB, FDN e FDA, lignina, celulose, cafeína, Ca e P. Esse decréscimo ocorreu pelo fato da casca de café possuir teores inferiores ao da polpa para PB, FDN, FDA, cafeína, Ca e P. Houve também redução linear nos valores de N-NH3, e somente a silagem com maior porcentagem de casca de café obteve valor inferior a 10%, considerado como limite para silagem de boa qualidade. A produção de efluentes foi maior para a silagem de polpa de café sem casca e não se obteve nenhuma produção na silagem com 60% de casca. Não houve diferença significativa na DIVMS entre os tratamentos. A casca de café foi efetiva em aumentar o teor de MS da silagem de PC e em reduzir os teores de FDN, FDA, N-NH3, proporcionando silagem de valor nutritivo satisfatório à alimentação de bovinos, criando, assim, alternativa para aproveitamento desses resíduos. Ao considerar o teor de MS encontrado nas silagens, a melhor quantidade de casca de café a ser acrescentada à polpa de café, para produção de silagem está entre 30% e 35%.
This study aimed to evaluate the chemical composition and fermentation characteristics of the coffee pulp silages with different proportions of coffee hulls. The material was ensiled in PVC with 150 mm diameter by 750 mm high, according to the treatments: coffee pulp (CoP), CoP + 20% of coffee hulls (CH), CoP + 40% of CH and CoP + 60% CH in a completely randomized design with six replications. The silos were opened 60 days after closing, when samples were taken for determination of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, cellulose, pH, N-NH3, caffeine, calcium (Ca), phosphorus (P) and in vitro digestibility of dry matter. We also determined gas production and effluent. There was a linear increase in DM content and pH, and a linear decrease of CP, NDF and ADF, lignin, cellulose, caffeine, Ca and P. This reduction occurs because the coffee hulls have lower levels than pulp to CP, NDF, ADF, caffeine, Ca and P. There was also a linear reduction in N-NH3 values, and only the highest percentage of silage with coffee hulls obtained below 10%, considered as the limit for good quality silage. Effluent production was higher for silage shelled coffee pulp and do not get any production in silage with 60% coffee hull. There was no significant difference in vitro digestibility of dry matter among treatments. The coffee hulls was effective in increasing DM content of CoP silage and to reduce NDF, ADF, N-NH3, providing nutritional value of silage satisfactory for cattle feed, creating an alternative for recovery of such waste. When considering the DM content found in silages, the amount of bark best coffee to be added to coffee pulp for the production of silage is between 30% and 35%.
Assuntos
Bovinos , Café , Medicago sativa , Silagem , Fermentação , Valor NutritivoRESUMO
Este trabalho teve como objetivo avaliar a composição química e as características fermentativas de silagens da polpa de café com diferentes proporções de casca de café. O material foi ensilado em PVC de 150 mm de diâmetro por 750 mm de altura, segundo os tratamentos: polpa de café (PC), PC + 20% de casca de café (CC), PC + 40% de CC e PC + 60% de CC, segundo delineamento inteiramente casualizado com seis repetições. Os silos foram abertos 60 dias após o fechamento, quando foram retiradas amostras para determinação dos teores de matéria seca (MS), proteína bruta (PB), fibra em detergente neutro (FDN), fibra em detergente ácido (FDA), lignina, celulose, pH, N-NH3, cafeína, cálcio (Ca), fósforo (P) e digestibilidade in vitro da MS (DIVMS). Determinou-se também a produção de efluentes e de gás. Houve um aumento linear no teor de MS e pH, à medida que foi adicionado casca de café nas silagens, e decréscimo linear nos teores de PB, FDN e FDA, lignina, celulose, cafeína, Ca e P. Esse decréscimo ocorreu pelo fato da casca de café possuir teores inferiores ao da polpa para PB, FDN, FDA, cafeína, Ca e P. Houve também redução linear nos valores de N-NH3, e somente a silagem com maior porcentagem de casca de café obteve valor inferior a 10%, considerado como limite para silagem de boa qualidade. A produção de efluentes foi maior para a silagem de polpa de café sem casca e não se obteve nenhuma produção na silagem com 60% de casca. Não houve diferença significativa na DIVMS entre os tratamentos. A casca de café foi efetiva em aumentar o teor de MS da silagem de PC e em reduzir os teores de FDN, FDA, N-NH3, proporcionando silagem de valor nutritivo satisfatório à alimentação de bovinos, criando, assim, alternativa para aproveitamento desses resíduos. Ao considerar o teor de MS encontrado nas silagens, a melhor quantidade de casca de café a ser acrescentada à polpa de café, para produção de silagem está entre 30% e 35%. (AU)
This study aimed to evaluate the chemical composition and fermentation characteristics of the coffee pulp silages with different proportions of coffee hulls. The material was ensiled in PVC with 150 mm diameter by 750 mm high, according to the treatments: coffee pulp (CoP), CoP + 20% of coffee hulls (CH), CoP + 40% of CH and CoP + 60% CH in a completely randomized design with six replications. The silos were opened 60 days after closing, when samples were taken for determination of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, cellulose, pH, N-NH3, caffeine, calcium (Ca), phosphorus (P) and in vitro digestibility of dry matter. We also determined gas production and effluent. There was a linear increase in DM content and pH, and a linear decrease of CP, NDF and ADF, lignin, cellulose, caffeine, Ca and P. This reduction occurs because the coffee hulls have lower levels than pulp to CP, NDF, ADF, caffeine, Ca and P. There was also a linear reduction in N-NH3 values, and only the highest percentage of silage with coffee hulls obtained below 10%, considered as the limit for good quality silage. Effluent production was higher for silage shelled coffee pulp and do not get any production in silage with 60% coffee hull. There was no significant difference in vitro digestibility of dry matter among treatments. The coffee hulls was effective in increasing DM content of CoP silage and to reduce NDF, ADF, N-NH3, providing nutritional value of silage satisfactory for cattle feed, creating an alternative for recovery of such waste. When considering the DM content found in silages, the amount of bark best coffee to be added to coffee pulp for the production of silage is between 30% and 35%. (AU)
Assuntos
Bovinos , Medicago sativa , Silagem , Café , Fermentação , Valor NutritivoRESUMO
An analytical method based on capillary electrophoresis (CE) with a partially aqueous electrolyte system was developed to enable the free fatty acids of Brazil nut oil to be identified in cosmetic formulations. In this study, a gel cream formulation was developed and its oil phase was extracted with a mixture of chloroform-methanol-water (1:2:0.8 v/v/v). The chloroform layer was saponified with a methanolic solution of NaOH (0.5 mol L-1) at 75-80 °C for 25 minutes. Experiments were carried out on a Beckman PACE/MDQ CE system (Fullerton, CA, USA) equipped with an on-column, diode-array detection system set at 254 nm and at 25ºC. The electrolyte consisted of 12.5 mmol L-1 sodium tetraborate buffer pH 7.0, 12.5 mmol L-1 polyoxyethylene 23-lauryl ether, 7.5 mmol L-1 sodium dodecylbenzenesulfonate (used as chromophore for indirect UV detection) and acetonitrile (35% v/v). The proposed method allowed the separation and identification of the fatty acids of Brazil nut oil in a cosmetic gel cream, as well as enabling possible interference by the oily phase components in the formulation to be identified.
Um método de análise por eletroforese capilar com sistema de eletrólito parcialmente aquoso foi desenvolvido para identificar os ácidos graxos livres do óleo de Castanha do Brasil em formulações cosméticas. No presente trabalho foi desenvolvida uma formulação cosmética (gel creme) cuja fase oleosa foi extraída com uma mistura de clorofórmio-metanol-água (1:2:0.8 v/v/v). A camada de clorofórmio, foi saponificada com solução de NaOH em metanol (0,5 mol L-1) a 75-80 °C durante 25 minutos. Os experimentos foram realizados em sistema de eletroforese capilar Beckman PACE/MDQ (Fullerton, CA, USA), com detecção de arranjo de diodos a 254 nm e a 25 ºC. O eletrólito utilizado foi 12,5 mmol L-1 de tetraborato de sódio tampão a pH 7,0, 12,5 mmol L-1 de éter de polioxietileno 23-lauril, 7,5 mmol L-1 de dodecilbenzenosulfonato de sódio (utilizado como agente cromóforo para detecção UV indireta) e acetonitrila (35% v/v). O método proposto permitiu a separação e a identificação dos ácidos graxos do óleo de Castanha do Brasil em formulações cosméticas, bem como possibilitou a identificação de interferências presentes na fase oleosa da formulação.
Assuntos
Cosméticos , Eletroforese Capilar , Ácidos GraxosRESUMO
Here we present a sensitive and specific liquid chromatography-tandem mass spectrometric method for the quantification of dimenhydrinate (I) in human plasma. Sample preparation is conducted using citalopram (II) addition as an internal standard (IS), liquid-liquid extraction with basified plasma using a mixture hexane/acetate (1:1, v/v) as the extracting solvent, and the final extract reconstituted in the mobile phase. I and II (IS) were injected in a C8 column with the mobile phase composed of methanol:isopropanol:water:formic acid (78.00:19.92:2.00:0.08, v/v/v/v) and monitored using a positive electrospray source with tandem mass spectrometry analyses. The selected reaction monitoring (SRM) was set using precursor ion and product ion combinations of m/z 256.0>167.0 and m/z 325.0>109.0 for I and II, respectively. The limit of quantification (LOQ) was 0.4 ng/mL, the dynamic range being 0.4-200 ng/mL. Validation results on linearity, specificity, accuracy, precision and stability, as well as on application to the analysis of plasma samples taken up to 24 h after oral administration of 100 mg of dimenhydrinate in healthy volunteers demonstrated its applicability to bioavailability studies.
Assuntos
Cromatografia Líquida/métodos , Dimenidrinato/sangue , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Disponibilidade Biológica , Dimenidrinato/química , Dimenidrinato/farmacocinética , Antagonistas dos Receptores Histamínicos H1/sangue , Antagonistas dos Receptores Histamínicos H1/química , Antagonistas dos Receptores Histamínicos H1/farmacocinética , Humanos , Estrutura Molecular , Reprodutibilidade dos TestesRESUMO
AIM: The aim of this study was to investigate factors that may affect the evolution of the caloric prescription in critically ill patients. Local: Intensive care unit patients. PATIENTS: 60 patients (33 M and 27 F); median age = 49 (1593) y were followed prospectively. They were divided in three groups according to the diagnostic: (a) trauma (n=20); (b) surgical (n=22), and 3) medical treatment (n=18). Forty-and-one (68.3%) patients received enteral nutrition (EN), 17 (28.3%) parenteral nutrition (TPN), and 2 (3.4%) TNP and EN. Nutritional status was graded B or C by global subjective evaluation. METHODS: Endpoints of the study were the time to begin the nutritional support, success or failure of the caloric prescription, and the evolution of the planned caloric prescription. The caloric evolution was considered as success if the prescription for the patient attained: (a) 25% of the caloric requirements on the 1st day; (b) 50% until the 3rd day; (c) 75% until the 6th day; and (e) 100% until the 10th day of the beginning of the support. RESULTS: In 54 (90%) patients, the nutritional support has begun until 48 h after admission and in 73.3% (44 patients), until the first 24 hours. EN was most prescribed for both trauma and medical patients while NPT was most used for surgical patients (p < 0.01). Success in caloric prescription was obtained in 73.3% (44) of the patients. There was no statistical difference for the success on the evolution of the prescription related to sex, age, diagnostic group, albumin level, type of support, mortality, use of fiber or glutamine. Success was attained earlier in patients without (median = 3.8 [95% CI, 5.7-16.7] days) than with (11.2 [95% CI, 5.7-16.7] days; p < 0.01) mechanical ventilation. CONCLUSIONS: Early nutritional support and success on the evolution of the caloric prescription can be accomplished in most critically ill patients. Evolution of the caloric prescription was slower in mechanical ventilated patients.
Assuntos
Estado Terminal , Nutrição Enteral , Terapia Nutricional , Nutrição Parenteral , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil , Interpretação Estatística de Dados , Ingestão de Energia , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Estado Nutricional , Estudos Prospectivos , Respiração Artificial , Fatores de TempoRESUMO
AIM: Peroxisome-proliferator-activated receptor-gamma2 (PPAR-gamma2) is a nuclear receptor that plays an important role in lipid metabolism and insulin sensitivity. The purpose of this study is to investigate the association of Pro12Ala polymorphism at the PPAR-gamma2 gene in Brazilian patients with type-2 diabetes mellitus (T2Dm) and controls (CG). METHODS: Genomic DNA was obtained from 207 unrelated white people presenting with T2Dm and from 170 controls. Anthropometric data included body mass index and waist to hip ratio. Biochemical parameters included fasting plasma glucose, total cholesterol, high- and low-density lipoprotein cholesterol, triglycerides, glycated haemoglobin and insulin. Systolic and diastolic blood pressures were also measured. Screening for mutations in the entire coding region of the PPAR-gamma gene was performed by means of polymerase chain reaction (PCR), single-strand conformational polymorphism and sequencing. Pro12Ala polymorphism was analysed by using PCR-RFLP (restriction fragment-length polymorphism). RESULTS: One base substitution was identified - a C to G substitution in exon B of the PPAR-gamma2 gene. The frequency of the Ala12 allele in T2Dm (0.09) was similar to that found in CG (0.06, p = 0.185). In the T2Dm group, Ala12 allele was associated with lower fasting plasma insulin levels (p = 0.036) and higher insulin sensitivity (p = 0.049) by means of homeostasis model assessment. Among obese people, there was no association between any of the T2Dm or obesity-related traits and the Pro12Ala polymorphism. CONCLUSIONS: The results of our study suggest that people with the Ala12 allele of the PPAR-gamma2 gene could be more sensitive to insulin than those carriers of the Pro12 allele among Brazilian Caucasians.
Assuntos
Diabetes Mellitus Tipo 2/genética , Resistência à Insulina/genética , PPAR gama/genética , Polimorfismo Genético , Adulto , Idoso , Antropometria , Glicemia/metabolismo , Análise Mutacional de DNA/métodos , Diabetes Mellitus Tipo 2/sangue , Feminino , Humanos , Insulina/sangue , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita SimplesRESUMO
AIM: The aim of the present study was to examine the effects of the C161T polymorphism of the peroxisome proliferator-activated receptor gamma (PPARgamma) gene in Brazilian subjects with Type 2 diabetes mellitus (T2DM) and controls residing in Sao Paulo City, Brazil. METHODS: Genomic DNA was obtained from 207 patients with T2DM and 170 unrelated normoglycemic individuals (CG). Anthropometric data included: body mass index, waist, hip, waist-to-hip ratio; biochemical parameters: fasting plasma glucose, total cholesterol, HDL- and LDL-cholesterol, triglycerides, glycated hemoglobin and insulin. Systolic and diastolic blood pressure was also measured. Screening for mutations in the entire coding region of the PPARgamma gene was carried out by PCR, single strand conformational polymorphism analysis (SSCP) and sequencing. C161T polymorphism was analyzed by PCR-RFLP. RESULTS: The C161T polymorphism was the only variant found in exon 6 of the PPARgamma gene. The frequency of the 161T allele in T2DM (0.10) was similar to that found in CG (0.07, p=0.210). Serum triglycerides (p=0.040), VLDL-cholesterol (p=0.040) and Atherogenic Index of Plasma (AIP; p=0.003) were significantly lower in 161T allele carriers than non-carriers in women of the T2DM group. CONCLUSIONS: Our results show that the C161T polymorphism in the PPARgamma gene is not associated with variables related to T2DM or insulin resistance in the Brazilian population. However, a reduction of serum triglycerides and AIP was observed in women with 161T allele of the C161T polymorphism of the PPARgamma gene.