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1.
Forensic Sci Int ; 306: 110056, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31765883

RESUMO

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.


Assuntos
Osso e Ossos/química , Impressões Digitais de DNA , Família , Dente/química , Restos Mortais , Sepultamento , Cromossomos Humanos Y , DNA/isolamento & purificação , Exumação , Feminino , Genética Forense , Sequenciamento de Nucleotídeos em Larga Escala , História do Século XX , Humanos , Masculino , Repetições de Microssatélites , Linhagem , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Eslovênia , II Guerra Mundial
2.
J Colloid Interface Sci ; 559: 215-225, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31629275

RESUMO

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.


Assuntos
Aminas/química , Carbono/química , Complexos de Coordenação/química , Cobre/química , Chumbo/química , Adsorção , Animais , Osso e Ossos/química , Cinética , Propriedades de Superfície , Suínos , Termodinâmica , Água , Poluentes Químicos da Água/química
3.
Sci Total Environ ; 700: 134470, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31693958

RESUMO

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.


Assuntos
Osso e Ossos/química , Carvão Vegetal/química , Cobre/química , Adsorção , Animais , Bovinos , Cinética , Modelos Químicos , Resíduos Sólidos
4.
J Agric Food Chem ; 68(3): 759-768, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31841328

RESUMO

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.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Anti-Hipertensivos/química , Osso e Ossos/química , Gelatina/química , Peptídeos/química , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Animais , Anti-Hipertensivos/administração & dosagem , Anti-Hipertensivos/isolamento & purificação , Pressão Sanguínea/efeitos dos fármacos , Bovinos , Humanos , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Masculino , Simulação de Acoplamento Molecular , Peptídeos/administração & dosagem , Peptídeos/isolamento & purificação , Peptidil Dipeptidase A/química , Hidrolisados de Proteína/química , Ratos , Ratos Endogâmicos SHR
5.
Forensic Sci Int ; 305: 110027, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31704515

RESUMO

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.


Assuntos
Osso e Ossos/química , Proteínas do Citoesqueleto/isolamento & purificação , Proteínas da Matriz Extracelular/isolamento & purificação , Fogo , Animais , Anticorpos/análise , Biomarcadores/química , Western Blotting , Técnica de Desmineralização Óssea , Osso e Ossos/patologia , Colágeno/ultraestrutura , Proteínas do Citoesqueleto/imunologia , Eletroforese , Proteínas da Matriz Extracelular/imunologia , Patologia Legal/métodos , Humanos , Microscopia Eletrônica de Varredura , Suínos , Temperatura Ambiente
6.
Forensic Sci Int ; 305: 110031, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31707238

RESUMO

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.


Assuntos
Degradação Necrótica do DNA , DNA/análise , Preservação Biológica/métodos , Manejo de Espécimes/métodos , Água , Osso e Ossos/química , Humanos , Unhas/química , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Saliva/química , Temperatura Ambiente , Fatores de Tempo
7.
Mater Sci Eng C Mater Biol Appl ; 104: 109966, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31499942

RESUMO

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.


Assuntos
Apatitas/química , Osso e Ossos/química , Nanopartículas/química , Carbonatos/química , Cristalografia/métodos , Microscopia Eletrônica de Transmissão/métodos
8.
Isotopes Environ Health Stud ; 55(5): 478-498, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31402710

RESUMO

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.


Assuntos
Isótopos de Carbono/análise , Fezes/química , Isótopos de Nitrogênio/análise , Panthera/fisiologia , Comportamento Predatório , Animais , Belize , Osso e Ossos/química , Carnivoridade/fisiologia , Colágeno/análise , Colágeno/química , Ecossistema , Isótopos de Estrôncio/análise
9.
Molecules ; 24(16)2019 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-31431001

RESUMO

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.


Assuntos
Osso e Ossos/química , Quitosana/química , Animais , Regeneração Óssea/efeitos dos fármacos , Quitina/química , Humanos , Osteogênese/efeitos dos fármacos , Polímeros/química , Engenharia Tecidual/métodos , Tecidos Suporte
10.
Forensic Sci Int Genet ; 42: 268-274, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31442669

RESUMO

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.


Assuntos
Restos Mortais , Sepultamento , Impressões Digitais de DNA , DNA Mitocondrial/genética , Adolescente , Adulto , Osso e Ossos/química , Criança , Feminino , Genoma Humano , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala , História Medieval , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência de DNA , Suécia , Dente/química , Adulto Jovem
11.
J Anim Sci ; 97(10): 4023-4031, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31376282

RESUMO

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.


Assuntos
Osso e Ossos/química , Calcificação Fisiológica/fisiologia , Trato Gastrointestinal/fisiologia , Fósforo na Dieta/administração & dosagem , Suínos/fisiologia , Ração Animal/análise , Animais , Dieta/veterinária , Digestão , Feminino , Masculino , Ossos Metacarpais/química , Necessidades Nutricionais , Fósforo na Dieta/metabolismo , Distribuição Aleatória , Suínos/crescimento & desenvolvimento
12.
Sci Total Environ ; 696: 133965, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31461692

RESUMO

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.


Assuntos
Bactérias/metabolismo , Biodegradação Ambiental , Fosfatos/metabolismo , Fósforo/metabolismo , Osso e Ossos/química , Cinética , Fósforo/química , Microbiologia do Solo , Solubilidade
13.
Forensic Sci Int Genet ; 42: 181-189, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31374457

RESUMO

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.


Assuntos
Cromossomos Humanos Y , Degradação Necrótica do DNA , Impressões Digitais de DNA , Haplótipos , Reação em Cadeia da Polimerase/métodos , Osso e Ossos/química , Eletroforese Capilar , Genética Forense/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Repetições de Microssatélites , Análise de Sequência de DNA , Dente/química
14.
ScientificWorldJournal ; 2019: 7060847, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31346324

RESUMO

The design of bone scaffolds for tissue regeneration is a topic of great interest, which involves different issues related to geometry of architectures, mechanical behavior, and biological requirements, whose optimal combination determines the success of an implant. Additive manufacturing (AM) has widened the capability to produce structures with complex geometries, which should potentially satisfy the different requirements. These architectures can be obtained by means of refined methods and have to be assessed in terms of geometrical and mechanical properties. In this paper a triply periodic minimal surface (TPMS), the Schwarz's Primitive surface (P-surface), has been considered as scaffold unit cell and conveniently parameterized in order to investigate the effect of modulation of analytical parameters on the P-cell geometry and on its properties. Several are the cell properties, which can affect the scaffold performance. Due to the important biofunctional role that the surface curvature plays in mechanisms of cellular proliferation and differentiation, in this paper, in addition to properties considering the cell geometry in its whole (such as volume fraction or pore size), new properties were proposed. These properties involve, particularly, the evaluation of local geometrical-differential properties of the P-surface. The results of this P-cell comprehensive characterization are very useful for the design of customized bone scaffolds able to satisfy both biological and mechanical requirements. A numerical structural evaluation, by means of finite element method (FEM), was performed in order to assess the stiffness of solid P-cells as a function of the changes of the analytical parameters of outer surface and the thickness of cell. Finally, the relationship between stiffness and porosity has been analyzed, given the relevance that this property has for bone scaffolds design.


Assuntos
Células Artificiais , Regeneração Óssea , Osso e Ossos/química , Engenharia Tecidual , Tecidos Suporte/química , Algoritmos , Células Artificiais/química , Substitutos Ósseos/química , Modelos Estruturais , Porosidade , Propriedades de Superfície , Engenharia Tecidual/métodos
15.
Poult Sci ; 98(9): 3611-3616, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31321433

RESUMO

Laying hen skeletal health continues to be an industry priority. Bone ash and bone Ca quantification in laying hen long bones provides valuable information on skeletal health. Unfortunately, these measurements can only be accomplished by sacrificing hens, thus making longitudinal measurements on the same hen impossible. Quantitative computed tomography (QCT), used with a calcium hydroxyapatite phantom, has been used to determine bone density of wings and legs as well as live hens throughout the production cycle by scanning with a calcium hydroxyapatite phantom. QCT has also been used to scan live hens throughout the production cycle. The purpose of this study was to determine how QCT calculated bone mineral content (QCT BMC) corresponds to analytical bone Ca and bone ash. Wing and leg quarters from 72-wk-old W-36 hens were QCT scanned along with a QCT Phantom. After scanning, humeri, femurs, and tibias were cleaned, divided into epiphysis (E) and diaphysis (D), fat extracted, ashed, and digested under nitric acid, and Ca was determined by atomic absorption spectroscopy was used to determine E, D, and whole bone Ca. Four bones/type were used for E and D, while 6 bones/type were used for whole bone measurements. A second set of bones were prepared to determine correlation of BMC to bone ash. QCT scans were analyzed with Mimics software (Materialise NV, Leuven, Belgium) to calculate bone volume and density in Hounsfield units. Utilizing the QCT phantom and bone volume, BMC was calculated for E, D, and whole bone. Data were analyzed with regression analysis and Pearson correlation coefficients were determined. Analytical Ca was correlated to QCT BMC for E (R = 0.84, P < 0.01), D (R = 0.99, P < 0.01), and whole bone (R = 0.97, P < 0.01). Whole bone ash was highly correlated to QCT BMC for femur (N = 47, R = 0.97, P < 0.001), tibia (N = 50, R = 0.94, P < 0.001), and keel (N 50, R = 0.94, P < 0.001). Whole bone ash and QCT BMC values of femur and tibia were not different (P = 0.39 and 0.22 respectively). Based on this information, QCT could provide relative quantitative assessment of total bone mineral in live birds proving useful in long-term studies.


Assuntos
Densidade Óssea/fisiologia , Osso e Ossos/química , Cálcio/metabolismo , Galinhas/fisiologia , Minerais/análise , Animais , Feminino , Fêmur/química , Úmero/química , Esterno/química , Tíbia/química , Tomografia Computadorizada por Raios X/veterinária
16.
BMC Musculoskelet Disord ; 20(1): 330, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307450

RESUMO

BACKGROUND: The aetiology of bone marrow oedema-like abnormalities (BMOA) seen on magnetic resonance imaging (MRI) is as yet not fully understood. The current study aimed to investigate the potential of projection radiography and Raman microspectroscopy to provide information regarding the underlying physiological changes associated with BMOA in equine bone samples. METHODS: MRI was used to assess 65 limbs from 43 horses. A subset of 13 limbs provided 25 samples, 8 with BMOA present and 17 as controls; these were examined with projection radiography to assess bone mineral density and Raman spectroscopy to assess bone composition. Statistical analysis was conducted using SPSS, the relationship between BMOA and age was tested using binary logistic regression, other outcome measures via unpaired t-tests. RESULTS: Overall BMOA was found to be associated with locally increased bone density (p = 0.011), suggesting increased bone formation; however, no measurable changes relating to bone remodelling were found, and there were no detectable changes in the chemical composition of bone. CONCLUSIONS: BMOA is associated with locally increased bone density, without an associated change in the chemical composition of bone, suggesting this is not linked to BMOA. The presence of increased bone density associated with BMOA does appear to suggest that an increased amount of bone formation is occurring in these regions, but as Raman microspectroscopy data do not demonstrate any significant changes in bone chemical composition associated with BMOA, it would appear that the increased bone volume is due to a greater amount of bone being formed rather than an imbalance in relation to bone remodelling. The study provides a proof of principle for the use of Raman microspectroscopy and projection radiography in in vitro studies of BMOA.


Assuntos
Densidade Óssea , Doenças Ósseas/veterinária , Doenças da Medula Óssea/veterinária , Osso e Ossos/química , Edema/veterinária , Animais , Doenças Ósseas/diagnóstico por imagem , Doenças Ósseas/patologia , Doenças da Medula Óssea/diagnóstico por imagem , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/patologia , Edema/diagnóstico por imagem , Edema/patologia , Membro Anterior , Cavalos , Imagem por Ressonância Magnética , Espectroscopia de Ressonância Magnética/métodos , Estudo de Prova de Conceito , Análise Espectral Raman/métodos
17.
Ecotoxicol Environ Saf ; 182: 109400, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31276883

RESUMO

This study was designed to investigate skeletal changes in Podocnemis unifilis embryos derived from artificially incubated eggs exposed to different concentrations of atrazine, glyphosate or atrazine and glyphosate mixture. Forty-two eggs were randomly allocated to one of seven trays containing vermiculite treated distilled water (control group) or the following solutions: 2 or 200 µg L-1 of atrazine (groups A1 and A2 respectively); 65 or 6500 µg L-1 of glyphosate (groups G1 and G2 respectively); 2 µg L-1 and 65 µg L-1 or 200 µg L-1 and 6500 µg L-1 of atrazine and glyphosate mixture (groups AG1 and AG2 respectively). Three eggs per tray were randomly collected on days 30 and 50 of the incubation period. Embryos were submitted to soft tissue diaphanization and stained with Alizarin red S or Alcian blue for morphological analysis of bone and cartilage tissues; histological analysis was performed to confirm ossification changes. Findings were compared between groups. Morphological changes were limited to sclerotic ring features and number of ribs. Malformations rates differed significantly (p < 0.05) between embryos in the control and treated groups A2, AG1 and AG2. Concurrent exposure to atrazine and glyphosate did not affect the presence or severity of embryonic malformations and was not associated with appendicular skeleton changes in P. unifilis embryos. However, further studies focusing on the axial skeleton with particular emphasis on rib abnormalities are warranted.


Assuntos
Atrazina/toxicidade , Glicina/análogos & derivados , Óvulo/efeitos dos fármacos , Praguicidas/toxicidade , Testes de Toxicidade , Animais , Atrazina/análise , Desenvolvimento Ósseo , Osso e Ossos/química , Cartilagem , Ovos , Glicina/toxicidade , Osteogênese , Tartarugas
18.
Food Chem ; 299: 125103, 2019 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-31284247

RESUMO

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.


Assuntos
Osso e Ossos/química , Galinhas , Proteínas de Aves Domésticas/química , Animais , Emulsões/química , Interações Hidrofóbicas e Hidrofílicas , Peso Molecular , Papaína/química , Proteínas de Aves Domésticas/isolamento & purificação , Solubilidade , Temperatura Ambiente , Fatores de Tempo , Ultrassom/métodos , Resíduos
19.
Molecules ; 24(13)2019 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-31261852

RESUMO

A method based on gas chromatography-mass spectrometry (GC-MS) is described for the determination of bisoprolol and atenolol in human bone. After the addition of lobivolol as internal standard, pulverized samples were incubated in acetonitrile for 1 h under ultrasounds. After adjusting the pH of the samples to 6, they were centrifuged, and the supernatants were subjected to solid phase extraction. Elution was achieved by using 3 mL of 2% ammonium hydroxide in 80:20 dichloromethane:isopropanol solution. Eluted samples were evaporated and derivatized. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.1-0.3 ng/mg (depending on the drug) to 150 ng/mg, the mean absolute recoveries were 60% for bisoprolol and 106% for atenolol, the matrix effect was 69% for bisoprolol and 70% for atenolol and process efficiency was 41% for bisoprolol and 80% for atenolol. The intra- and inter-assay accuracy values were always better than 12%. The validated method was then applied to bone samples from two real forensic cases in which toxicological analysis in blood were positive for atenolol in the first case (0.65 µg/mL) and bisoprolol in the second case (0.06 µg/mL). Atenolol was found in bone samples from the corresponding case at the approximate concentration of 148 ng/mg and bisoprolol was found at 8 ng/mg.


Assuntos
Atenolol/análise , Bisoprolol/análise , Osso e Ossos/química , Toxicologia Forense , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Reprodutibilidade dos Testes , Extração em Fase Sólida
20.
Poult Sci ; 98(10): 4982-4993, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31152669

RESUMO

This study evaluated the effects of total sulfur amino acid (TSAA) levels on performance, egg quality, and bone metabolism in laying hens subjected or not to high environmental temperature (HT). HyLine W36 layers (n = 144) were randomly distributed in a 2 × 3 factorial arrangement. Room temperature (control, CR: 21°C/24 h; and high temperature, HR: 32°C/8 h) and diets (70, 85, or 100% of TSAA) were the main factors, with 4 replicates of 6 birds (19 to 45 wk). The TSAA levels were obtained by adding L-Methionine (L-Met) to the basal diet (70% of TSAA) until 85 and 100% of TSAA were reached. At weeks 21, 34, and 45, growth performance, egg production, and egg quality traits were evaluated. At 45 wk, bones were evaluated for collagenous and non-collagenous proteins, bone volume, mineral content, and mineral density from total, cortical, trabecular, and medullary portions. When interactions were found, the increase of TSAA levels (85 and 100%) was able to counteract the negative effects of HT. In general, HT reduced egg production (P < 0.05) and did not significantly affect bone quality. The birds fed 70% of TSAA showed higher feed conversion, lower body weight, egg weight, and egg mass than birds fed 85 and 100% of TSAA in at least one phase. The birds fed 100% of TSAA showed higher egg production and egg mass than the other treatments at 21 wk of age. The cortical and trabecular bone mineral densities were higher for birds fed 100 than 70% of TSAA, whereas the medullary bone mineral content and density were higher for birds fed 70 than 100% of TSAA. In conclusion, HT had negative impact on performance, egg quality and no effect on bone development. The supplementation of L-Met until either 85 or 100% of TSAA levels were reached was enough to assure good performance, egg quality, and bone development in laying hens subjected or not to HT.


Assuntos
Osso e Ossos/química , Galinhas/fisiologia , Temperatura Alta , Metionina/metabolismo , Óvulo/fisiologia , Ração Animal/análise , Animais , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Feminino , Metionina/administração & dosagem , Distribuição Aleatória
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