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1.
J Agric Food Chem ; 68(7): 2091-2098, 2020 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-31927882

RESUMEN

In this study, we used reversed-phase high performance liquid chromatography (LC) to isolate three novel peptides with calcium-chelating capacity from tilapia bone collagen hydrolysate. Using LC-tandem mass spectrometry, we determined the amino acid sequences to be GPAGPHGPVG, FDHIVY, and YQEPVIAPKL. We then synthesized the three peptides and verified their calcium-chelating activity. Results showed that the calcium-chelating activity of GPAGPHGPVG, FDHIVY, and YQEPVIAPKL reached 18.80 ± 0.49, 35.73 ± 0.74, and 28.4 ± 0.94 mg/g, respectively. We next investigated how each peptide enhanced intestinal calcium absorption using Caco-2 cell monolayers. Compared with the control group, GPAGPHGPVG, FDHIVY, and YQEPVIAPKL potently enhanced calcium transport within 30 min by 89 ± 9, 202 ± 12, and 130 ± 7%, respectively. Results suggest that these peptides isolated from tilapia bone hydrolysate can be used as dietary supplements to increase calcium absorption.


Asunto(s)
Huesos/química , Calcio/química , Calcio/metabolismo , Quelantes/química , Colágeno/química , Proteínas de Peces/química , Mucosa Intestinal/metabolismo , Péptidos/química , Secuencia de Aminoácidos , Animales , Transporte Biológico/efectos de los fármacos , Células CACO-2 , Humanos , Mapeo Peptídico , Hidrolisados de Proteína/química , Tilapia
2.
Forensic Sci Int ; 306: 110056, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31765883

RESUMEN

The killings during the Second World War, with nearly one hundred thousand victims, is one of the greatest losses of life in Slovenia's modern history. This article presents the genetic identification of the victims of the largest family massacre that occurred in Slovenia, in which 10 members of the same family were killed. Seven of them were buried in a hidden mass grave and only two children survived. In 2015 and 2016, two graves were found and three incomplete female skeletons and at least three incomplete male skeletons were exhumed. A total of 12 bones and teeth were analysed and compared to two living relatives. Extracted DNA was quantified using the PowerQuant kit, and various autosomal and Y-STR kits were used for STR typing. Up to 2.7 ng DNA/g of powder was acquired from the samples analysed. We managed to obtain nuclear DNA for successful STR typing from seven bones and one molar. From the female grave, autosomal profiles were obtained only from one skeleton, and from the male grave from five out of six femurs. The relationships between the males were additionally confirmed by analyses of Y-STRs. STR profiles made possible the identification of four family members; one of the aunts from the female grave, and two uncles and the father of the surviving children, who were used as family references, from the male grave. The product rule was used to calculate a combined likelihood ratio for autosomal and Y-STRs, and statistical analyses showed high confidence of correct identification with posterior probability (PP) greater than 99.9 % for three out of four victims identified. For identifying the aunt, the PP obtained after ESI-17 and NGM STR typing was too low. To increase the PP, the next-generation sequencing Precision ID GlobalFiler NGS STR Panel was used and, after the analysis of additional STR loci, the statistical analysis showed a PP greater than 99.9 %, indicating that a sufficient number of genetic markers had been investigated in identifying the skeletal remains of the aunt. An elimination database containing the genetic profiles of all individuals that had been in contact with the bones was created to ensure traceability in case of contamination, and no matches were found. After more than 70 years, the skeletal remains were returned to the surviving children, who buried their relatives in a family grave.


Asunto(s)
Huesos/química , Dermatoglifia del ADN , Familia , Diente/química , Restos Mortales , Entierro , Cromosomas Humanos Y , ADN/aislamiento & purificación , Exhumación , Femenino , Genética Forense , Secuenciación de Nucleótidos de Alto Rendimiento , Historia del Siglo XX , Humanos , Masculino , Repeticiones de Microsatélite , Linaje , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Eslovenia , Segunda Guerra Mundial
3.
Anal Bioanal Chem ; 412(2): 259-265, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31776641

RESUMEN

Synchrotron radiation micro-X-ray fluorescence (SR-µXRF) is a powerful elemental mapping technique that has been used to map tungsten and zinc distribution in bone tissue. However, the heterogeneity of the bone samples along with overlap of the tungsten L-edge with the zinc K-edge signals complicates SR-µXRF data analysis, introduces minor artefacts into the resulting element maps, and decreases image sensitivity and resolution. To confirm and more carefully delineate these SR-µXRF results, we have employed laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to untangle the problem created by the K/L-edge overlap of the tungsten/zinc pair. While the overall elemental distribution results are consistent between the two techniques, LA-ICP-MS provides significantly higher sensitivity and image resolution compared with SR-µXRF measurements in bone. These improvements reveal tissue-specific distribution patterns of tungsten and zinc in bone, not observed using SR-µXRF. We conclude that probing elemental distribution in bone is best achieved using LA-ICP-MS, though SR-µXRF retains the advantage of being a non-destructive method with the capability of being paired with X-ray techniques, which determine speciation in situ. Since tungsten is an emerging contaminant recently found to accumulate in bone, accurately determining its distribution and speciation in situ is essential for directing toxicological studies and informing treatment regimes. Graphical abstract Tungsten and zinc localization and uptake in mouse femurs were imaged by synchrotron radiation, left, and by laser ablation ICP-MS, right. The increased resolution of the LA-ICP-MS technique resolves the problem of the overlap in tungsten's L-edge and zinc's K-edge.


Asunto(s)
Huesos/química , Rayos Láser , Espectrometría por Rayos X/métodos , Tungsteno/análisis , Zinc/análisis , Animales , Masculino , Espectrometría de Masas/métodos , Ratones , Ratones Endogámicos C57BL , Sincrotrones
4.
Anal Bioanal Chem ; 412(1): 27-36, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31705221

RESUMEN

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and laser-induced breakdown spectroscopy (LIBS) are widely accepted techniques for direct sampling of biological materials for elemental analysis, with increasing applications being reported over the recent years. This review is focused on the calibration materials used to quantify trace elements in different biological samples such as soft tissues (for instance brain, liver, hair) and hard tissues (bones and teeth). The design of a correct calibration strategy relies on the choice of an adapted reference material that can be commercially available or prepared in-house, which will be reviewed here. A large variety of methods has been approached and considered promising over the years, and the development of matrix-matched reference biological materials seems now closer than ever and gives hope to even better quantitation using LIBS and LA-ICP-MS.


Asunto(s)
Espectrometría de Masas/métodos , Oligoelementos/análisis , Huesos/química , Encéfalo/metabolismo , Calibración , Cabello/química , Humanos , Hígado/química , Estándares de Referencia , Diente/química , Oligoelementos/normas
5.
J Agric Food Chem ; 68(3): 759-768, 2020 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-31841328

RESUMEN

In this study, we investigated the antihypertensive effects in vitro and in vivo of novel angiotensin-converting enzyme inhibitory (ACEI) peptides purified and identified from bovine bone gelatin hydrolysate (BGH). Thirteen ACEI peptides were identified from BGH, and among which, RGL-(Hyp)-GL and RGM-(Hyp)-GF exhibited high ACE inhibition with IC50 values of 1.44 and 10.23 µM. Molecular docking predicted that RGM-(Hyp)-GF and ACE residues of Glu384, His513, and Lys511 formed hydrogen-bonding interactions at distances of 2.57, 2.99, and 2.42 + 3.0 Å. RGL-(Hyp)-GL formed hydrogen bonds with Lys511 and Tyr523 and generated hydrogen-bonding interactions with His387 and Glu411 in the zinc(II) complexation motif at distances of 2.74 and 3.03 + 1.93 Å. The maximal decrements in systolic blood pressure in spontaneously hypertensive rats induced by one-time gavage of RGL-(Hyp)-GL and RGM-(Hyp)-GF at 30 mg/kg were 31.3 and 38.6 mmHg. RGL-(Hyp)-GL had higher enzyme degradation resistance than that of RGM-(Hyp)-GF in vitro incubation in rat plasma, and they were sequentially degraded into pentapeptides and tetrapeptides within 2 h. Our results indicate that BGH can serve as a nutritional candidate to control blood pressure.


Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/química , Antihipertensivos/química , Huesos/química , Gelatina/química , Péptidos/química , Inhibidores de la Enzima Convertidora de Angiotensina/administración & dosificación , Inhibidores de la Enzima Convertidora de Angiotensina/aislamiento & purificación , Animales , Antihipertensivos/administración & dosificación , Antihipertensivos/aislamiento & purificación , Presión Sanguínea/efectos de los fármacos , Bovinos , Humanos , Hipertensión/tratamiento farmacológico , Hipertensión/fisiopatología , Masculino , Simulación del Acoplamiento Molecular , Péptidos/administración & dosificación , Péptidos/aislamiento & purificación , Peptidil-Dipeptidasa A/química , Hidrolisados de Proteína/química , Ratas , Ratas Endogámicas SHR
6.
J Colloid Interface Sci ; 559: 215-225, 2020 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-31629275

RESUMEN

This work aimed to develop a feasible technology to reuse dead pig biowaste for heavy metals removal, which was first carbonized into pig bone char (PBC) after pyrolysis and further functionalized with amino (NH2) to improve its affinity with heavy metals. The application of PBC after NH2-functionalization for aqueous heavy metals removal was explored, including Pb2+, Cu2+, and different complex Cu(II) species. PBC@SiO2-NH2 showed comparable performance with AC@SiO2-NH2. The adsorption capacity of Pb2+ and Cu2+ by PBC@SiO2-NH2 was 120 and 30 mg g-1, respectively. The NH2 functionalization enhanced the adsorption of complex Cu(II) species, and PBC@SiO2-NH2 exhibited good performance under a wide pH range and coexisting ions. The adsorption of Cu(II)EDTA species with an EDTA/Cu(II) molar ratio of 1.0 by PBC would be enhanced ~3 times and ~6 times after NH2 functionalization and further addition of Fe3+, respectively. The results of EDX, elemental mapping, and XPS confirmed the adsorption of Cu(II) species. The adsorption kinetics, isotherm, and thermodynamics of different Cu(II) species by PBC@SiO2-NH2 were investigated. The regeneration of PBC@SiO2-NH2 was easily performed via acidic wash. These results suggested the application potential of the reuse of biowaste, which could be a promising adsorbent for aqueous heavy metals after specific functionalization.


Asunto(s)
Aminas/química , Carbono/química , Complejos de Coordinación/química , Cobre/química , Plomo/química , Adsorción , Animales , Huesos/química , Cinética , Propiedades de Superficie , Porcinos , Termodinámica , Agua , Contaminantes Químicos del Agua/química
7.
Sci Total Environ ; 700: 134470, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31693958

RESUMEN

With the increasing demand for beef worldwide, a considerable amount of bovine bone is discharged as solid waste. Therefore, in this study, the physicochemical properties of chars from bovine bones (ribs, scapulae, vertebrae, and legs) and their copper sorption behavior in aqueous solutions were investigated. The bone chars were pyrolyzed at 500 °C and the ash contents were approximately 85.08%, although the leg bone char had significantly higher values. The rib bone char showed a larger specific surface area (172 m2/g), smaller average pore diameter (7.7 nm), and more basic functional groups than the other char types. The maximum sorption capacity varied from 72.53 to 83.71 g/kg, with the rib bone char exhibiting the best adsorption characteristics, followed by the scapulae, vertebrae, and legs. A correlation analysis demonstrated that the adsorption capacity of Cu(II) on bone char is closely related to surface pore characteristics. An adsorption kinetic analysis and physicochemical characterization of the chars indicate that the Cu(II) adsorption mechanism in bovine bone char is primarily surface chemisorption. Based on the different of physicochemical properties and sorption behavior, bone chars pyrolyzed from bovine ribs are most suitable for adsorption-related applications. The results of this study demonstrate the potential for classified utilization of animal bones, including the use of graded bone chars as low-cost adsorbents requiring no chemical pre-treatment.


Asunto(s)
Huesos/química , Carbón Orgánico/química , Cobre/química , Adsorción , Animales , Bovinos , Cinética , Modelos Químicos , Residuos Sólidos
8.
Mar Pollut Bull ; 143: 175-186, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31789153

RESUMEN

To examine whether historical fish bones can record the magnitude of heavy metal pollution, we analyzed up to 700 years old fish bone remains extracted from an ornithogenic sediment profile on Nandao Island, South China Sea. Bulk sediments and subfossil fish bones were analyzed for elemental and mineralogical composition, as well as stable carbon and nitrogen isotopes. The results showed that pre-1850 CE fish bones experienced significant diagenesis, and could not be used to reconstruct historical record of heavy metal pollution. Fish bone diagenesis was mainly attributed to the erosion from guano in sediment profile. In contrast, the fish bones from in post-1850 CE time were well preserved and could provide useful information on historical pollution loads over the past 160 years. Since 1850 CE, relatively high concentrations of heavy metals from anthropogenic sources, especially Zn, were recorded in fish bone subfossils on Nandao Island, South China Sea.


Asunto(s)
Huesos/química , Productos Pesqueros , Fósiles , Metales Pesados/análisis , Contaminantes Químicos del Agua/análisis , Animales , Radioisótopos de Carbono/análisis , China , Monitoreo del Ambiente/métodos , Sedimentos Geológicos , Islas , Isótopos de Nitrógeno , Espectroscopía Infrarroja por Transformada de Fourier
9.
Forensic Sci Int ; 305: 110031, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31707238

RESUMEN

A rectrospective study was conducted on the effect of the long term storage of 122 DNA samples resuspended in water, one of the elution media still suggested by well established protocols. These DNA samples come from four different kinds of forensically relevant samples (saliva swabs, FTA card bloodstains, nails and II° World War bones) extracted in 2008-2018 and stored at - 20°C (n=113 of groups #1-#5) and at +4°C (n=9 of the group #6), respectively. At the time of the present study (2019), quantitative PCR (qPCR) was employed as tool for assessing the degradation of the samples. The employment of the Human Quantifiler Kit showed that the median loss of DNA ranged from 17.8% to 66.6% in groups #1-#5 while it was 85.0% in group #6. However, it is likely that these values represent an underestimation due to the shortness of the qPCR probe (62 bp). Noteworthy, the DNA loss was statistically significant in each of the six groups (p values ≤ 0.0167). Thus, in agreement with the data on spontaneous DNA decay, no forensic DNA sample should be stored in water for long term periods. In conclusion, the results of this technical note warn against the use of water for this purpose.


Asunto(s)
Degradación Necrótica del ADN , ADN/análisis , Preservación Biológica/métodos , Manejo de Especímenes/métodos , Agua , Huesos/química , Humanos , Uñas/química , Reacción en Cadena de la Polimerasa , Estudios Retrospectivos , Saliva/química , Temperatura Ambiental , Factores de Tiempo
10.
Forensic Sci Int ; 305: 110027, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31704515

RESUMEN

Due the proteins from bone remains are highly resistant to pass of time and environmental conditions, they could tell us about the events that probably happened in the past. In the forensic and physical anthropology context, burnt bone remains are one of the most common pieces of recovered evidence and, generally, they are associated with funerary practices, criminal scenes or massive catastrophic events. In the present study, bone pieces of pigs were calcined at different calcination temperatures, and proteins were searched using biochemical, immunochemical and ultrastructure visualization under these experimentally conditions. For this purpose, it was successfully developed a non-demineralizing protein extraction method from burnt bone remains and the use of specific antibodies permitted the identification of different extracellular matrix and intracellular proteins. While collagen proteins type I and IV were identified and detected under middle and high calcination temperatures (300°C and 600°C); cytoskeletal proteins as actin, tubulin and, the microtubule associated protein Tau, were found under calcination process, even up high calcination temperatures. Under ultrastructural analysis, fibrous materials with a classical disposition of collagens were observed even at high calcination temperatures of the burnt bone remains. The protein identification and characterization in burnt bones as performed in present studies, is clearly demonstrating that using specific strategies for protein characterizations it is possible to found protein biomarkers in burnt bone remains and this strategy could be useful for forensic and anthropological purposes.


Asunto(s)
Huesos/química , Proteínas del Citoesqueleto/aislamiento & purificación , Proteínas de la Matriz Extracelular/aislamiento & purificación , Fuego , Animales , Anticuerpos/análisis , Biomarcadores/química , Western Blotting , Técnica de Desmineralización de Huesos , Huesos/patología , Colágeno/ultraestructura , Proteínas del Citoesqueleto/inmunología , Electroforesis , Proteínas de la Matriz Extracelular/inmunología , Patologia Forense/métodos , Humanos , Microscopía Electrónica de Rastreo , Porcinos , Temperatura Ambiental
11.
Analyst ; 144(20): 6128-6135, 2019 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-31535118

RESUMEN

Bone is one of the main sample types used for building chronologies in archaeology. It is also used in other research areas such as palaeodiet and palaeoenvironmental studies. However, for results to be accurate, samples must be free of exogenous carbon. Contamination can originate from a wide range of sources in the post-depositional environment but may also occur during excavation and post excavation activities (i.e. with the application of conservation materials) or during laboratory handling. Efficient procedures to remove contamination are therefore crucial prior to radiocarbon or stable isotope measurements. This work describes the development of an innovative sample pretreatment for bones, based on using supercritical CO2, which shows unique solvation properties. The effectiveness of supercritical fluid extraction (SFE) to remove conservation materials was compared with that obtained when applying a routine extraction based on the use of organic solvents (methanol, acetone and chloroform). The chemical composition of the bone samples before and after the two pre-treatments was then investigated using analytical pyrolysis-based techniques: EGA-MS (Evolved Gas Analysis-Mass Spectrometry) and Py-GC/MS (Pyrolysis-Gas Chromatography coupled with Mass Spectrometry). Collagen samples extracted from the same bones, prepared with the two cleaning protocols, were also radiocarbon dated by Accelerator Mass Spectrometry (AMS). The results of this study show that SFE is an efficient alternative method because it was as effective as the established treatment protocol. It removes contaminants such as conservation materials from bone samples with a minimum of handling and can be used routinely in radiocarbon dating laboratories. This work also demonstrates that analytical pyrolysis is not only a very efficient method to identify contaminants in bones but also to assess the effectiveness of the pretreatment prior to the radiocarbon measurement of the samples.


Asunto(s)
Arqueología , Huesos/química , Dióxido de Carbono/química , Radioisótopos de Carbono/análisis , Colágeno/aislamiento & purificación , Descontaminación/métodos , Nitrógeno/análisis , Cromatografía con Fluido Supercrítico , Cromatografía de Gases y Espectrometría de Masas , Humanos , Datación Radiométrica
12.
Mater Sci Eng C Mater Biol Appl ; 105: 110014, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31546414

RESUMEN

Cuttlefish bone (CB) has been explored as biomaterial in the bone tissue-engineering field due to its unique porous structure and capacity of the aragonite mineral to be hydrothermally converted into calcium phosphates (CaPs). In the present study, undoped and ion (Sr2+, Mg2+ and/or Zn2+) doped biphasic calcium phosphate (BCP) scaffolds were prepared by hydrothermal transformation (HT, 200 °C, 24 h) of CB. The obtained scaffolds were sintered and then coated with two commercial polymers, poly(ε-caprolactone) (PCL) or poly(DL-lactide) (PDLA), and with two synthesized ones, a poly(ester amide) (PEA) or a poly(ester urea) (PEU) in order to improve their compressive strength. The scaffolds were characterized by X-ray diffraction (XRD) coupled with structural Rietveld refinement, Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM). The results demonstrate that CB could be entirely transformed into BCPs in the presence or absence of doping elements. The initial CB structure was preserved and the polymeric coatings did not jeopardize the interconnected porous structure. Furthermore, the polymeric coatings enhanced the compressive strength of the scaffolds. The in vitro bio-mineralization upon immersing the scaffolds into simulated body fluid (SBF) demonstrated the formation of bone-like apatite surface layers in both uncoated and coated scaffolds. Overall, the produced scaffolds exhibit promising properties for bone tissue engineering applications.


Asunto(s)
Huesos/química , Fosfatos de Calcio/farmacología , Materiales Biocompatibles Revestidos/farmacología , Decapodiformes/anatomía & histología , Polímeros/farmacología , Andamios del Tejido/química , Animales , Huesos/ultraestructura , Calcificación Fisiológica , Fuerza Compresiva , Módulo de Elasticidad , Porosidad , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Temperatura Ambiental , Difracción de Rayos X
13.
Mater Sci Eng C Mater Biol Appl ; 104: 109966, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31499942

RESUMEN

In this study we present the first crystal structure model for bone apatite based on the analysis of individual nanocrystals by high resolution transmission electron microscopy (HRTEM). Crystallographic image processing of the obtained HRTEM images from different projections indicates symmetry reduction with respect to P63/m stoichiometric apatites and the presence of threefold symmetry along the c axis. Based on HRTEM observations and the measured Ca/P = 2 ratio we propose a structural model with phosphate-to-carbonate substitution and O vacancies localized along c axis, which explains the observed loss of 63 screw axis parallel, and the shift of mirror plane perpendicular to the c axis. Also, the presence of non-equivalent (010) surfaces has been proven. These results on the atomic structure of bone apatite nanocrystals contribute to the understanding of their biochemically controlled nucleation processes.


Asunto(s)
Apatitas/química , Huesos/química , Nanopartículas/química , Carbonatos/química , Cristalografía/métodos , Microscopía Electrónica de Transmisión/métodos
14.
J Anim Sci ; 97(10): 4023-4031, 2019 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-31376282

RESUMEN

A study was conducted to determine the standardized total tract digestible phosphorus (STTD P) requirement for 24- to 130-kg finishing pigs housed under commercial conditions. A total of 1,130 barrows and gilts (PIC 359 × 1050, Hendersonville, TN; initially 24.2 kg) were used, with 26 to 27 pigs per pen with 7 replicates per treatment. Pens of pigs were allotted to treatments in a randomized complete block design with body weight (BW) as the blocking factor. The dietary treatments were fed in 4 phases and were formulated to contain 80%, 90%, 100%, 115%, 130%, and 150% of the National Research Council (NRC) requirement estimate for finishing pigs within each phase. Weight ranges for each phase were: 27 to 49, 49 to 76, 76 to 90, and 90 to 130 kg. Treatments were achieved by increasing the amount of monocalcium phosphate at the expense of corn in the diet with no added phytase. All diets were formulated to contain a similar 1.14:1 to 1.16:1 total Ca:P ratio across treatments in all phases. Increasing STTD P resulted in a quadratic response (P < 0.05) in average daily gain (ADG), gain-to-feed ratio (G:F), and final BW. The greatest improvement was observed with STTD P at 130% of NRC for ADG and final BW and at 115% STTD P for G:F. Average daily feed intake increased linearly (linear, P < 0.05) with the inclusion of STTD P. Increasing STTD P resulted in an increase (quadratic, P < 0.05) in hot carcass weight (HCW) and carcass ADG with the greatest response observed with STTD P at 130% of NRC. There was a marginally significant response (quadratic, P < 0.10) in carcass G:F, with the greatest improvement with STTD P at 115% of NRC. Carcass yield decreased (linear, P < 0.05) with increasing STTD P, while there was a marginally significant (linear, P < 0.10) decrease in backfat and increase in fat-free lean. At the end of the study, a metacarpal was collected and analyzed for bone ash. Increasing STTD P resulted in an increase (linear, P < 0.05) in bone ash weight and percentage ash. For ADG and G:F, the quadratic model demonstrated the best fit. The maximum response in ADG and G:F was estimated at 122% and 116% of NRC STTD P, respectively. The broken-line linear model best fit the data for percentage bone ash, with a plateau achieved at 131% of the NRC STTD P. In conclusion, the estimated STTD P requirement of 24 to 130 kg ranged from 116% to 131% of the NRC publication (2012) requirement estimate.


Asunto(s)
Huesos/química , Calcificación Fisiológica/fisiología , Tracto Gastrointestinal/fisiología , Fósforo Dietético/administración & dosificación , Porcinos/fisiología , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Digestión , Femenino , Masculino , Huesos del Metacarpo/química , Necesidades Nutricionales , Fósforo Dietético/metabolismo , Distribución Aleatoria , Porcinos/crecimiento & desarrollo
15.
Forensic Sci Int Genet ; 42: 181-189, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31374457

RESUMEN

Forensic and population genetics often rely on Y-chromosomal studies. Whether it is a human identification case, trace evidence examination or phylogenetic analysis, a Y-STR haplotype is an important tool in the hands of law enforcement agencies. A common obstacle in achieving satisfactory results in all of the above mentioned circumstances, is low DNA quantity and quality within samples obtained. In this study we have examined Y-STR haplotypes in 75 bone material samples, coming from different time periods. For this purpose we have chosen YFiler Plus PCR Amplification Kit (ThermoFisher Scientific) and ForenSeq Signature DNA Prep Kit (Verogen Inc.), which use two different allele calling technologies - capillary electrophoresis and Massively Parallel Sequencing respectively. Full profiles were obtained from DNA extracts with as little as 0.1896 ng (Degradation Index 1.3) (ForenSeq) and 0.0591 ng (Degradation Index 26.8) (YFiler Plus) DNA input. The results that we present in this paper show differences in amplification rates between common markers in both kits. The differences strictly reflect mean amplicon length of markers. This, however, does not seem to influence Y-haplogroup estimation results noticeably. In one sample a discordance occurred between haplotypes obtained with both methods, where a 24 allele was called in DYS390 marker by capillary electrophoresis, while for the same sample in this locus a 23 allele was shown with MPS. A reason for this is yet to be investigated. The sequence analysis revealed a significant variation between isometric alleles, especially within repetitive regions of studied Y-STR markers.


Asunto(s)
Cromosomas Humanos Y , Degradación Necrótica del ADN , Dermatoglifia del ADN , Haplotipos , Reacción en Cadena de la Polimerasa/métodos , Huesos/química , Electroforesis Capilar , Genética Forense/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Repeticiones de Microsatélite , Análisis de Secuencia de ADN , Diente/química
16.
Forensic Sci Int Genet ; 42: 268-274, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31442669

RESUMEN

In 1998, a Viking Age mass grave was discovered and excavated at St. Laurence´s churchyard in Sigtuna, Sweden. The excavated bones underwent osteoarchaeological analysis and were assigned to at least 19 individuals. Eleven skeletons showed sharp force trauma from bladed weapons. Mass graves are an unusual finding from this time period, making the burial context extraordinary. To investigate a possible maternal kinship among the individuals, bones and teeth from the skeletal remains were selected for mitochondrial DNA (mtDNA) analysis. Sanger sequencing of short stretches of the hypervariable segments I and II (HVS-I and HVS-II) was performed. A subset of the samples was also analysed by massively parallel sequencing analysis (MPS) of the entire mtDNA genome using the Precision ID mtDNA Whole Genome Panel. A total of 15 unique and three shared mtDNA profiles were obtained. Based on a combination of genetic and archaeological data, we conclude that a minimum of 20 individuals was buried in the mass grave. The majority of the individuals were not maternally related. However, two possible pairs of siblings or mother-child relationships were identified. All individuals were assigned to West Eurasian haplogroups, with a predominance of haplogroup H. Although the remains showed an advanced level of DNA degradation, the combined use of Sanger sequencing and MPS with the Precision ID mtDNA Whole Genome Panel revealed at least partial mtDNA data for all samples.


Asunto(s)
Restos Mortales , Entierro , Dermatoglifia del ADN , ADN Mitocondrial/genética , Adolescente , Adulto , Huesos/química , Niño , Femenino , Genoma Humano , Haplotipos , Secuenciación de Nucleótidos de Alto Rendimiento , Historia Medieval , Humanos , Masculino , Persona de Mediana Edad , Análisis de Secuencia de ADN , Suecia , Diente/química , Adulto Joven
17.
Molecules ; 24(16)2019 Aug 19.
Artículo en Inglés | MEDLINE | ID: mdl-31431001

RESUMEN

Chitosan is a deacetylated polysaccharide from chitin, the natural biopolymer primarily found in shells of marine crustaceans and fungi cell walls. Upon deacetylation, the protonation of free amino groups of the d-glucosamine residues of chitosan turns it into a polycation, which can easily interact with DNA, proteins, lipids, or negatively charged synthetic polymers. This positive-charged characteristic of chitosan not only increases its solubility, biodegradability, and biocompatibility, but also directly contributes to the muco-adhesion, hemostasis, and antimicrobial properties of chitosan. Combined with its low-cost and economic nature, chitosan has been extensively studied and widely used in biopharmaceutical and biomedical applications for several decades. In this review, we summarize the current chitosan-based applications for bone and dental engineering. Combining chitosan-based scaffolds with other nature or synthetic polymers and biomaterials induces their mechanical properties and bioactivities, as well as promoting osteogenesis. Incorporating the bioactive molecules into these biocomposite scaffolds accelerates new bone regeneration and enhances neovascularization in vivo.


Asunto(s)
Huesos/química , Quitosano/química , Animales , Regeneración Ósea/efectos de los fármacos , Quitina/química , Humanos , Osteogénesis/efectos de los fármacos , Polímeros/química , Ingeniería de Tejidos/métodos , Andamios del Tejido
18.
Isotopes Environ Health Stud ; 55(5): 478-498, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31402710

RESUMEN

Faecal isotopic analysis may complement other non-invasive wildlife survey tools for monitoring landscape use by carnivores, such as motion-detecting cameras and non-invasive genetic sampling. We analysed carbon, nitrogen, and strontium isotopes in faecal matter produced by jaguars (Panthera onca) as well as bones from consumed prey at the Mountain Pine Ridge Forest Reserve (MPR) in Belize, Central America. The MPR is ideally suited for a spatial isotope study as vegetation and geology both vary considerably. The isotopic composition of faecal matter should reflect the habitat and geology where consumed prey lived. We used bone from consumed prey recovered from jaguar scats as a proxy for diet. Faecal matter and bone showed comparable spatial isotopic trends, suggesting that the isotopic composition of jaguar faeces can be used to detect foraging in different habitats (pine forest versus broadleaf forest) or on different geologies (Mesozoic carbonates; Palaeozoic granite, contact metamorphics, and metasediments). This result is reassuring as bones are not always present in carnivore scats. Studying landscape use by cryptic and wide-ranging carnivore species like jaguars remains challenging. Isotopic analysis of faecal matter complements the existing array of non-invasive spatial monitoring tools.


Asunto(s)
Isótopos de Carbono/análisis , Heces/química , Isótopos de Nitrógeno/análisis , Panthera/fisiología , Conducta Predatoria , Animales , Belice , Huesos/química , Carnivoría/fisiología , Colágeno/análisis , Colágeno/química , Ecosistema , Isótopos de Estroncio/análisis
19.
Sci Total Environ ; 696: 133965, 2019 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-31461692

RESUMEN

The enhanced phosphorus (P) release from charred bone by microorganisms results in hotspots to alleviate P limitation in agricultural and natural systems. This study compared P release, assisted by phosphate-solubilizing bacteria (PSB), from charred bone (CB) produced at various temperatures (100-300 °C). In the absence of PSB, soluble P from CB in water was observed with fluctuation between 100 and 300 °C, with a maximum value of 8.66 mg/L at 200 °C. Similarly, kinetics of dissolution indicated that CB produced at 250 °C owned the highest solubility and dissolution rate. After the addition of PSB, soluble P from all the CB samples were all elevated. The CB produced at 100 °C incredibly showed the most significant enhancement (from 3.51 to 77.37 mg /L). ATR-IR and XPS confirmed the loss of organic matter (primarily collagen), but no significant mineralogical alternation of bioapatite in bone. Meanwhile, it demonstrated that collagen itself cannot provide soluble P. However, the collagen contributed to the substantial sorption of bacteria, which improved the efficiency of P release from CB surface. This study clarified the P release via the interaction between CB and PSB, and hence provided a new perspective on understanding P biogeochemical cycle in ecosystem.


Asunto(s)
Bacterias/metabolismo , Biodegradación Ambiental , Fosfatos/metabolismo , Fósforo/metabolismo , Huesos/química , Cinética , Fósforo/química , Microbiología del Suelo , Solubilidad
20.
Food Chem ; 299: 125103, 2019 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-31284247

RESUMEN

This study investigated the application of ultrasonic pretreatment (UP) to assist with enzymatic extraction of chicken bone protein. Relevant parameters of UP including ultrasonic power, duration of treatment and temperature were studied. The effect of ultrasound was evaluated by examining the protein structure changes (i.e. surface hydrophobicity, secondary and tertiary structures, molecular weight distribution) and their functionalities (i.e. solubility, oil holding capacity, foaming and emulsifying properties). Higher protein extraction yield and surface hydrophobicity were evident after UP, along with the changes in the secondary and tertiary structure of the protein. UP alone effects the proteins having molecule weight at about 100 kDa. Results also reveal UP was capable of enhancing the protein functionalities. Therefore, UP could be applied as a suitable technology to improve the yield and quality of the protein extracted from chicken bone by enzymatic method, and as such, facilitating the potential utilization of waste byproduct from poultry industry.


Asunto(s)
Huesos/química , Pollos , Proteínas de Aves de Corral/química , Animales , Emulsiones/química , Interacciones Hidrofóbicas e Hidrofílicas , Peso Molecular , Papaína/química , Proteínas de Aves de Corral/aislamiento & purificación , Solubilidad , Temperatura Ambiental , Factores de Tiempo , Ultrasonido/métodos , Residuos
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