Testing In Vivo

The determination of chemiluminescent intensity of reporter gene expression in vivo is generally disturbed by the presence of hemoglobin. Current methods consist in using perfusion to eliminate blood from investigated tumors or organs. In this work we propose a simple method to overcome this difficulty. The method consists in establishing an absorbance-dependence plot of the ratio R% ‫؍‬ ⌽/⌽ 0 between the chemiluminescent intensities measured when hemoglobin is present or absent. For every measurement of the luminescent intensity ⌽ on sample containing blood, if the absorbance A of the hemoglobin is determined, it allows one to have the intensity ratio R% which in turn gives the corrected intensity ⌽ 0 when the absorption by hemoglobin is eliminated. The method is particularly adapted for comparative measurements of transfection levels in tumors where perfusion cannot be easily performed. © 2002 Elsevier Science (USA)

Penetrating neural probe technologies allow investigators to record electrical signals in the brain. The implantation of probes causes acute tissue damage, partially due to vasculature disruption during probe implantation. This trauma can cause abnormal electrophysiological responses and temporary increases in neurotransmitter levels, and perpetuate chronic immune responses. A significant challenge for investigators is to examine neurovascular features below the surface of the brain in vivo. The objective of this study was to investigate localized bleeding resulting from inserting microscale neural probes into the cortex using two-photon microscopy (TPM) and to explore an approach to minimize blood vessel disruption through insertion methods and probe design. 3D TPM images of cortical neurovasculature were obtained from mice and used to select preferred insertion positions for probe insertion to reduce neurovasculature damage. There was an 82.8 ± 14.3% reduction in neurovascular damage for probes inserted in regions devoid of major (>5 µm) sub-surface vessels. Also, the deviation of surface vessels from the vector normal to the surface as a function of depth and vessel diameter was measured and characterized. 68% of the major vessels were found to deviate less than 49 µm from their surface origin up to a depth of 500 µm. Inserting probes more than 49 µm from major surface vessels can reduce the chances of severing major sub-surface neurovasculature without using TPM.

This article is a panel presentation by Professor Kathy Hessler in which she advocates for more critical attention paid to ethical and legal issues pertaining to animal testing. Notable topics of Professor Hessler’s talk include the philosophical underpinnings that have encouraged the use of animals in research testing, the scant patchwork of protections the legal system currently affords research animals, and the implications of shifting from in vivo to in vitro testing models, with the former predicated upon dualistic Cartesian concepts and the latter informed by the Darwinian paradigm. In discussing the moral issues related to animal testing, this article draws salient parallels between the denial of legal personhood to nonhuman animals with the same denial to women and minority groups. The article then concludes by comparing the American approach to animal testing with the shifting European Union approach, which affords considerably more weight to animal welfare concerns.

The osteocyte is believed to act as the primary sensor of mechanical stimulus in bone, controlling signalling for bone growth and resorption in response to changes in the mechanical demands placed on bones throughout life. Alterations in local bone tissue composition and structure arising during osteoporosis likely disrupt the local mechanical environment of these mechanosensitive bone cells, and may thereby initiate a mechanobiological response. However, due to the difficulties in directly characterising the mechanical environment of bone cells in vivo, the mechanical stimuli experienced by osteoporotic bone cells are not known. The global aim of this thesis is to discern the in vivo mechanical environment of the osteocyte, both in healthy bone tissue and during the disease of osteoporosis.
The first study of this thesis involved the development of 3D finite element models of osteocytes, including their cell body and the surrounding pericellular matrix (PCM) and extracellular matrix (ECM), using confocal images of the lacunar-canalicular network. These anatomically representative models demonstrated the significance of geometry for strain amplification within the osteocyte mechanical environment. A second study employed fluid-structure interaction (FSI) modelling to investigate the complex multi-physics environment of osteocytes in vivo. These models built upon the anatomically representative models developed in the first study, and FSI methods were used to simulate loading-induced interstitial fluid flow through the lacunar-canalicular network. Interestingly, the in vivo mechanical stimuli (strain and shear stress) predicted using these computational approaches were above thresholds known to elicit osteogenic responses from osteoblastic cells in vitro, and thereby provide a novel insight into the complex multi-physics mechanical environment of osteocytes in vivo.
The third study of this thesis sought to experimentally characterise the strain environment of osteoblasts and osteocytes under physiological loading conditions in healthy and osteoporotic bone, using a rat model of osteoporosis. A custom-designed loading device compatible with a confocal microscope was constructed to apply strains to fluorescently stained femur samples from normal and ovariectomised rats. Confocal imaging was performed simultaneously during loading and digital image
ii
correlation techniques were used to characterise cellular strains from the images acquired. These results suggested that the mechanical environment of osteoblasts and osteocytes is altered during early-stage osteoporosis, and it is proposed that a mechanobiological response restores the homeostatic mechanical environment by late-stage osteoporosis. A final study applied these results as inputs for the developed computational models to investigate whether changes in tissue properties, lacunar-canalicular architecture or amplification mechanisms during osteoporosis could explain the altered mechanical stimulation of osteocytes observed. The findings of this study shed new light on the osteocyte mechanical environment in both healthy and osteoporotic bone, elucidating a possible mechanobiological relationship between increases in strain stimulation of the osteocyte and subsequent increases in mineralisation of bone tissue as key events in the progression of osteoporosis.
Together, the studies in this thesis provide a novel insight into the closed mechanical environment of the osteocyte. Using both computational and experimental methods, the mechanical stimuli that osteocytes experience under physiological loading in vivo, in both healthy and osteoporotic bone, were elucidated. In particular, the research in this thesis provides a missing mechanobiological link in the temporal development of post-menopausal osteoporosis, and the information gained from this body of work may inform future treatments for osteoporosis.

The antioxidant potential (AP) is an important nutritional property of foods, as increased oxidative stress is involved in most diet-related chronic diseases. In dairy products, the protein fraction contains antioxidant activity, especially casein. Other antioxidants include antioxidant enzymes; lactoferrin; conjugated linoleic acid; coenzyme Q10; vitamins C, E, A and D3; equol; uric acid; carotenoids; and mineral activators of antioxidant enzymes. The AP of dairy has been extensively studied in vitro, with few studies in animals and humans. Available in vivo studies greatly differ in their design and objectives. Overall, on a 100 g fresh weight-basis AP of dairy is close to that of grain-based foods and vegetable or fruit juices. Among dairy, cheeses present the highest AP due to their higher protein content. AP of milk increases during digestion by up to 2.5 times because of released antioxidant peptides. AP of casein is linked to specific amino acids, whereas -lactoglobulin thiol groups play a major role in AP of whey. Thermal treatments such as UHT have no clear effect on AP of milk. Raw fat-rich milks have higher AP than less fat-rich milk, because of lipophilic antioxidants. Probiotic yoghurts and fermented milks have higher AP than conventional yoghurt and milk because proteolysis by probiotics releases antioxidant peptides. Among the probiotics, Lactobacillus casei/acidophilus leads to the highest AP. The data are insufficient for cheese, but fermentation-based changes appears to positively impact AP. In conclusion, AP might participate in the reported dairy product protective effects against some chronic diseases.

Aim: The phytochemical screening, antibacterial activities and in vivo toxicity of extracts of the leaves of scent leaf (Ocimum gratissimum L.) were investigated. Methods: All the analyses were carried out using standard scientific procedures using Soxhlet extraction, well-diffusion agar antimicrobial testing and in vivo acute toxicity testing. Results: The phytochemical analysis according to standard screening tests using conventional protocols revealed the presence of anthraquinone, saponins, tannins, terpenoids and alkaloids, which were detected in methanol extract analyzed. But, flavonoids, glycosides, phlobatannins and steroids were not detected in the methanol extracts analyzed. While only flavonoids were detected in chloroform extract. All other phytochemicals were absent. The extract fractions generally exhibited slight antibacterial activities on Staphylococcus aureus, Bacillus subtilis, Salmonella Typhi and Escherichia coli. But, the extracts showed no effect against Candida albicans. The minimum inhibitory concentration of O. gratissimum was determined with S. aureus and B. cereus recording MICs at the lowest concentration (12.5 mg/mL) of the methanol and chloroform extracts used. While methanol and chloroform extracts were found to have recorded moderate activity S. Typhi and E. coli at the MIC of 50mg/mL. The methanol and chloroform extract recorded MBC of 50 mg/mL on B. subtilis and S. Typhi. However, Salmonella Typhi was inhibited at the concentration of 100 mg/mL of chloroform scent leaf extract. The MICs of C. albicans were not determined in the methanol and chloroform scent leaf extracts analyzed. The in vivo toxicity of O. gratissimum extracts against albino rats revealed that the plant extracts were found to exhibit mild toxicity at higher doses, but the overall remark showed that the plant extract was safe at various concentrations. Conclusions: The plant can be used in the treatment of various diseases caused by the test microbes, and the plant has less toxicity when administered orally.

A B S T R A C T The toxic liver impairment caused by free radical injury or excessive reactive oxigen species (ROS) formation could be effectivelly attenuated by natural antioxidants. The present study aimed to explore and compare the hepatoprotective and antioxidant effects of free and encapsulated quercetin in in vitro and in vivo models of hepatotoxicity. Thus, quercetin was encapsulated in chitosan/alginate nanoparticles by gelation method. Both empty and quercetin-loaded nanoparticles revealed good safety profile in vitro, determined by the lack of cytotoxicity in human hepatoma HepG2 cells. The pretreatment of HepG2 cells with encapsulated quercetin (10 mg/ml) significantly attenuated the decrease in cell viability in H 2 ? 2-induced oxidative stress (0.1 mM H 2 ? 2) , thus showing an effective in vitro protection. In vivo evaluation of the antioxidant and hepatoprotective potential of free and encapsulated quercetin was performed in a model of paracetamol À induced liver injury in male Wistar rats. The oral pretreatment with encapsulated quercetin (0.18 mg/kg b.w., 7 days) significantly diminished the increased levels of serum transaminases ALT and AST, attenuated the lipid peroxidation and restored the levels of gluthation (a marker of cell antioxidant defence system). The protective effects of quercetin encapsulated in chitosan-based nanoformulation were superior to those of free quercetin. The results of the study suggest that the encapsulation of quercetin in chitosan/alginate nanoformulations might represent an effective therapeutic approach against oxidative stress induced liver injury.

Rapid prototyping is a valuable implant production tool that enables the investigation of individual geometric parameters, such as shape, porosity, pore size and permeability, on the biological performance of synthetic bone graft substitutes. In the present study, we have employed low-temperature direct 3D printing to produce brushite and monetite implants with different geometries. Blocks predominantly consisting of brushite with channels either open or closed to the exterior were implanted on the decorticated lumbar transverse processes of goats for 12 weeks. In addition, similar blocks with closed channel geometry, consisting of either brushite or monetite were implanted intramuscularly. The design of the channels allowed investigation of the effect of macropore geometry (open and closed pores) and osteoinduction on bone formation orthotopically. Intramuscular implantation resulted in bone formation within the channels of both monetite and brushite, indicating osteoinductivity of these resorbable materials. Inside the blocks mounted on the transverse processes, initial channel shape did not seem to significantly influence the final amount of formed bone and osteoinduction was suggested to contribute to bone formation. r

In the streptozotocin induced diabetic rats treated separately with aqueous, ethanol, acetone and chloroform extracts of the seeds of B. nigra, the increase in serum glucose value between 0 and 1 hr of glucose tolerance test (GTT) was the least (29 mg/dl) in aqueous extract treated animals while it was 54, 44 and 44mg/dl with chloroform, acetone and ethanol extracts respectively. In further studies carried out with aqueous extract, the effective dose was found to be 200 mg/kg body weight in GTT. Administration of 200 mg/kg body weight of aqueous extract to diabetic animals daily once for one month brought down fasting serum glucose (FSG) levels while in the untreated group FSG remained at a higher value. In the treated animals the increase in glycosylated hemoglobin (HbA1c) and serum lipids was much less when compared with the levels in untreated diabetic controls. These findings suggest that further studies with the aqueous extract of B. nigra seeds on its antidiabetic activity would be useful.

Galactoxyloglucan polysaccharide (PST001), isolated from the seed kernels of Tamarindus indica (Ti), was used both as reducing and capping agent for the preparation of gold nanoparticles (PST-Gold) of 20 nm size. The present study evaluated the anticancer effects of the PST-Gold nanoparticles both in vitro and in vivo. The cytotoxicity was evaluated in the murine cancer cell lines, Dalton's lymphoma ascites (DLA) and Ehrlich's ascites carcinoma (EAC). Galactoxyloglucan-gold nanoparticles (PST-Gold) not only retained the anticancer effects of PST001, but also showed enhanced cytotoxicity via induction of apoptosis even at lower doses and lesser incubation times. In vivo antitumor activity was tested in DLA and EAC murine ascites and EAC solid-tumor syngeneic mouse models. PST-Gold nanoparticles reduced tumor burden and increased median survival and life span significantly in both tumor models compared to the controls. The PST-Gold nanoparticles were very effective as a chemopreventive agent, showing the best overall response when administered prior to tumor induction. In the case of solid tumors, intratumoral administration of the PST-Gold nanoparticles yielded significant results with regard to survival and increment in lifespan as compared to intraperitoneal mode of drug administration. Further studies in higher animal models and in patients at high-risk for recurrence are warranted to fully explore and develop the potential of PST-Gold nanoconjugates as a chemopreventive and therapeutic anti-cancer agent.

The present study describes the effects of dopants of Mn and/or Ni on the structural, magnetic and in vivo/in vitro performances of β-tricalcium phosphate (β-TCP) bioceramics synthesized by a sol-gel method. Both types of dopants caused significant effects on the structure-related parameters of β-TCP, including the crystallite dimensions and lattice parameters. The magnetic properties of as-prepared samples were found to be affected by the type and amount of the dopant(s). The in vitro antimicrobial activities of doped β-TCPs against American Type Culture Collection (ATCC) reference bacterial and fungal strains were evaluated. Among them, only 1.35Ni-TCP displayed an effective antimicrobial activity against Gram-positive bacteria (50 μg/mL Minimum Inhibitory Concentration (MIC) range), while other types of doped β-TCPs did not possess with such effect. The in vivo tests showed that each formulation could be a good nominate for bone healing and bone treatment.

The development of technologies capable of early tumor detection is unquestionably demanded by physicians, as early diagnosis is key to achieve more efficient and less invasive treatments with improved outcomes. At the preclinical level, nanotechnology has already provided innovative solutions for tumor imaging and therapy, but it has failed to provide real early tumor diagnosis. In this work, an infrared nanothermometry‐based approach toward early melanoma detection, based on the changes produced in the thermal relaxation dynamics of tissues as the tumor develops, is introduced. In vivo experiments demonstrate that detection of incipient tumors from their very onset is possible through monitoring changes in their thermal relaxation dynamics using Ag2S infrared luminescent nanothermometers. For a total tumor development time of 14 days, luminescence nanothermometry allows tumor detection 6 days before its presence is evident by visual inspection. Simultaneous study of the tumoral vasculature reveals that the premature variation in the thermal relaxation dynamics is a consequence of the interplay between tumor angiogenesis and necrosis during the different tumor development stages.

Bioelectronics, electronic technologies that interface with biological systems, are experiencing rapid growth in terms of technology development and applications, especially in neuroscience and neuroprosthetic research. The parallel growth with optogenetics and in vivo multi-photon microscopy has also begun to generate great enthusiasm for simultaneous applications with bioelectronic technologies. However, emerging research showing artefact contaminated data highlight the need for understanding the fundamental physical principles that critically impact experimental results and complicate their interpretation. This review covers four major topics: (1) material dependent properties of the photoelectric effect (conductor, semiconductor, organic, photoelectric work function (band gap)); (2) optic dependent properties of the photoelectric effect (single photon, multiphoton, entangled biphoton, intensity, wavelength, coherence); (3) strategies and limitations for avoiding/minimizing photoelectric effects; and (4) advantages of and applications for light-based bioelectronics (photo-bioelectronics).

Quorum sensing (QS) is a key regulator of virulence factors and biofilm formation in Gram-negative bacteria such as Pseudomonas aeruginosa. Microorganisms that inhabit soil are of strategic importance in the discovery of compounds with anti-QS properties. The objective of the study was to test the culture extract of a taxonomically novel species of Paenibacillus strain 139SI for its inhibitory effects on the QS-controlled virulence factors and biofilm formation of Pseudomonas aeruginosa both in vitro and in vivo. The Paenibacillus sp. culture extract was used to test its anti-QS effects on the LasA protease, LasB elastase, pyoverdin production, and biofilm formation of P. aeruginosa as well as evaluate its therapeutic effects on lung bacteriology, pathology, hematological profile, and serum antibody responses of experimental animals in a rat model of chronic lung infection. Results showed significant decrease in the activities of QS-controlled LasA protease, LasB elastase pyoverdin, and biofilm formation of P. aeruginosa caused by the culture extract. Moreover, the extract significantly prolonged the survival times of rats and facilitated the clearance of biofilm infections from infected lungs. In conclusion, the antiquorum sensing effects of culture extract from a novel species of Paenibacillus provide new insights to combat biofilm-associated infections.

This article is a panel presentation by Professor Kathy Hessler in which she advocates for more critical attention paid to ethical and legal issues pertaining to animal testing. Notable topics of Professor Hessler’s talk include the philosophical underpinnings that have encouraged the use of animals in research testing, the scant patchwork of protections the legal system currently affords research animals, and the implications of shifting from in vivo to in vitro testing models, with the former predicated upon dualistic Cartesian concepts and the latter informed by the Darwinian paradigm. In discussing the moral issues related to animal testing, this article draws salient parallels between the denial of legal personhood to nonhuman animals with the same denial to women and minority groups. The article then concludes by comparing the American approach to animal testing with the shifting European Union approach, which affords considerably more weight to animal welfare concerns.

Implantable neural electrode technologies for chronic neural recordings can restore functional control to paralysis and limb loss victims through brain-machine interfaces. These probes, however, have high failure rates partly due to the biological responses to the probe which generates an inflammatory scar and subsequent neuronal cell death. L1 is a neuronal specific cell adhesion molecule and has been shown to minimize glial scar formation and promote electrode-neuron integration when covalently attached to the surface of neural probes. In this work, the acute microglial response to L1-coated neural probes was evaluated in vivo by implanting coated devices into the cortex of mice with fluorescently labeled microglia, and tracking microglial dynamics with multi-photon microscopy for the ensuing 6 h in order to understand L1's cellular mechanisms of action. Microglia became activated immediately after implantation, extending processes towards both L1-coated and uncoated control probes at similar velocities. After the processes made contact with the probes, microglial processes expanded to cover 47.7% of the control probes' surfaces. For L1-coated probes, however, there was a statistically significant 83% reduction in microglial surface coverage. This effect was sustained through the experiment. At 6 h post-implant, the radius of microglia activation was reduced for the L1 probes by 20%, shifting from 130.0 to 103.5 μm with the coating. Microglia as far as 270 μm from the implant site displayed significantly lower morphological characteristics of activation for the L1 group. These results suggest that the L1 surface treatment works in an acute setting by microglial mediated mechanisms.

Peptic ulcer is one of the worldwide diseases where 10% of adults are affected by peptic ulcers at least once in their lifetime. The goal of this study was to evaluate the effect of levan in treating peptic ulcer. The bacterial honey isolates called Bacillus sp. levan was utilized. Levan was chemically characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), 1 H and 13 C NMR analysis. Levan was used to treat gastric ulcers induced in rats by oral administration of 5 mL/kg ethanol. Microscopic examination of stomach sections indicated that treatment with 200 mg/kg levan effectively healed the ulcers. Levan had no antimicrobial activity against a common cause of ulcers such as Helicobacter pylori bacteria. Rather, we proposed that the high adhesion (manifested as a protective coating) and prebiotic activity of levan may account for the observed beneficial effects. The immunohistochemical examination showed that levan led to a noticeable Bacillus sp. levan reduction in NF-B in the upper gastric mucosa. The results concluded that the role of levan was more protective rather than preventive and suggested that levan could play a fundamental role in solving the peptic ulcer problems.

Intracortical neural probes enable researchers to measure electrical and chemical signals in the brain. However, penetration injury from probe insertion into living brain tissue leads to an inflammatory tissue response. In turn, microglia are activated, which leads to encapsulation of the probe and release of pro-inflammatory cytokines. This inflammatory tissue response alters the electrical and chemical microen-vironment surrounding the implanted probe, which may in turn interfere with signal acquisition. Dexamethasone (Dex), a potent anti-inflammatory steroid, can be used to prevent and diminish tissue disruptions caused by probe implantation. Herein, we report retrodialysis administration of dexa-methasone while using in vivo two-photon microscopy to observe real-time microglial reaction to the implanted probe. Microdialysis probes under artificial cerebrospinal fluid (aCSF) perfusion with or without Dex were implanted into the cortex of transgenic mice that express GFP in microglia under the CX3CR1 promoter and imaged for 6 h. Acute morphological changes in microglia were evident around the microdialysis probe. The radius of microglia activation was 177.1 mm with aCSF control compared to 93.0 mm with Dex perfusion. T-stage morphology and microglia directionality indices were also used to quantify the microglial response to implanted probes as a function of distance. Dexamethasone had a profound effect on the microglia morphology and reduced the acute activation of these cells.

In this work, untreated bovine cortical bones (BCBs) were exposed to a range of heat treatments in order to determine at which temperature the apatite develops an optimum morphology comprising porous nano hydroxyapatite (nanoHAp) crystals. Rectangular specimens (10 mm × 10 mm × 3-5 mm) of BCB were prepared, being excised in normal to longitudinal and transverse directions. Specimens were sintered at up to 900 °C under ambient pressure in order to produce apatites by two steps sintering. The samples were characterized by thermogravimetric analysis, X-ray diffraction (XRD), and scanning electron microscopy (SEM) attached to an energy-dispersive X-ray spectroscopy detector. For the first time, morphology of the HAp particles was predicted by XRD, and it was verified by SEM. The results show that an equiaxed polycrystalline HAp particle with uniform porosity was produced at 900 °C. It indicates that a porous nanoHAp achieved by sintering at 900 °C can be an ideal candidate as an in situ scaffold for load-bearing tissue applications. © 2013 by the authors; licensee MDPI, Basel, Switzerland.

http://www.mdpi.com/1996-1944/6/1/65

Aim: The phytochemical screening, antibacterial activities and in vivo toxicity of extracts of the leaves of scent leaf (Ocimum gratissimum L.) were investigated. Methods: All the analyses were carried out using standard scientific procedures using Soxhlet extraction, well-diffusion agar antimicrobial testing and in vivo acute toxicity testing. Results: The phytochemical analysis according to standard screening tests using conventional protocols revealed the presence of anthraquinone, saponins, tannins, terpenoids and alkaloids, which were detected in methanol extract analyzed. But, flavonoids, glycosides, phlobatannins and steroids were not detected in the methanol extracts analyzed. While only flavonoids were detected in chloroform extract. All other phytochemicals were absent. The extract fractions generally exhibited slight antibacterial activities on Staphylococcus aureus, Bacillus subtilis, Salmonella Typhi and Escherichia coli. But, the extracts showed no effect against Candi...

The evolution and subsequent design of clinically effective antima-larial drugs particularly 4-aminoquinolines, 8-aminoquinolines and 9-amino-acridines are reviewed. These molecules benefited from scientific advances in medicinal and synthetic chemistry guided by pharmacological screening, including animal models of malaria. The mechanism of action of antimalarials, especially against the heme receptor, and the impact of this knowledge on drug design is critically discussed. Modelling investigations and quantum mechanics calculations reveal close contacts between the porphyrin ring and selected atoms within compounds such as the bisquinoline, metaquine. Analysis of these close contacts can be used to design compounds with modulated antimalari-al activity to further clarify the drug action. Knowledge of mammalian drug metabolism and pharmacokinetics, together with detailed in vitro and in vivo pharmacology has aided (a) resurrection of old compounds and (b) redesign of existing compounds. The latter includes judicious modification (e.g. inversion of oxidizable functional groups as in SN-13,730 i.e. isoquine) or introduction of groups blocking metabolism (e.g. exploiting bond strength viz. fluorine or steric effects with the t-butyl group). This simultaneous modulation of both drug metabolism and interaction with the heme receptor can be used to enhance antimalarial activity. Compounds benefiting from such modifications include primaquine, pyronaridine, isoquine, metaquine and AQ-13, together with selected analogues with useful activity against drug-resistant Plasmodia in vi-vo. It is concluded that optimisation of the privileged quinoline and acridine scaffolds, discovered in the early part of the 20 th century, still has a vital role to play in the future discovery of cost effective solutions to malaria.

Different types of natural antioxidants (tocopherols -alone, in two different doses, or combined with ascorbic acid -and a rosemary extract) were employed in the production of corn flakes, to select the most effective ones for achieving low levels of volatile lipid oxidation products, related to off-flavours, after long-term storage. Samples were submitted to SPME/GC/MS and sensory analyses. Multivariate analysis elucidated the contribution of the volatile compounds to the sensory properties of corn flakes. Under the experimental conditions, tocopherols conferred better antioxidant activity than did rosemary extract, leading to low levels of off-flavour compounds, with a further improvement provided by the synergistic activity of ascorbic acid. This could be attributed to interfacial and phase partitioning phenomena involving antioxidants, depending on their lipo-or hydrophilicity, in heterogeneous food products. Acids, n-alkanols, lactones and some aldehydes appeared to have a considerable influence on the intensity of the off-flavours.

Different types of natural antioxidants (tocopherols -alone, in two different doses, or combined with ascorbic acid -and a rosemary extract) were employed in the production of corn flakes, to select the most effective ones for achieving low levels of volatile lipid oxidation products, related to off-flavours, after long-term storage. Samples were submitted to SPME/GC/MS and sensory analyses. Multivariate analysis elucidated the contribution of the volatile compounds to the sensory properties of corn flakes. Under the experimental conditions, tocopherols conferred better antioxidant activity than did rosemary extract, leading to low levels of off-flavour compounds, with a further improvement provided by the synergistic activity of ascorbic acid. This could be attributed to interfacial and phase partitioning phenomena involving antioxidants, depending on their lipo-or hydrophilicity, in heterogeneous food products. Acids, n-alkanols, lactones and some aldehydes appeared to have a considerable influence on the intensity of the off-flavours.

Creative teamwork in multidisciplinary teams is a topic of interest to cognitive psychologists on the one hand, and to both social and organizational psychologists on the other. However, the interconnections between cognitive and social layers have been rarely explored. Drawing on mental models and dissonance theories, the current study takes a central variable studied by cognitive psychologists-analogy-and examines its relationship to a central variable examined by social psychologists-conflict. In an observational, field study, over 11 h of audio-video data from conversations of the Mars Exploration Rover scientists were coded for different types of analogy and micro-conflicts that reveal the character of underlying psychological mechanisms. Two different types of time-lagged logistic models applied to these data revealed asymmetric patterns of associations between analogy and conflict. Within-domain analogies, but not within-discipline or outside-discipline analogies, preceded science and work process conflicts, suggesting that in multidisciplinary teams, representational gaps in very close domains will be more likely to spark conflict. But analogies also occurred in reaction to conflict: Process and negative conflicts, but not task conflicts, preceded within-discipline analogies, but not to within-domain or outside-discipline analogies. This study demonstrates ways in which cognition can be bidirectionally tied to social processes and discourse.

Inflammatory bowel diseases (IBD) are immunomediated ailments affecting millions of individuals. Although diet is regarded as an important factor influencing IBD, there are no accepted dietary recommendations presently available. We administered 7·6 % lyophilised apples obtained from two cultivars (Golden Delicious and Marie Ménard, low and high in polyphenols, respectively) to HLA-B27 transgenic rats which develop spontaneous IBD. After 3 months feeding, rats fed Marie Ménard apples had reduced myeloperoxidase activity (3·6 (sem 0·3) v. 2·2 (sem 0·2) U/g tissue; P < 0·05) and reduced cyclo-oxygenase-2 (P < 0·05) and inducible NO synthase gene expression (P < 0·01) in the colon mucosa and significantly less diarrhoea (P < 0·05), compared with control rats. Cell proliferation in the colon mucosa was reduced significantly by feeding Golden Delicious apples, with a borderline effect of Marie Ménard apples. Gene expression profiling of the colon mucosa, analysed using the Whole Rat Genome 4 × 44 K Agilent Arrays, revealed a down-regulation of the pathways of PG synthesis, mitogen-activated protein kinase (MAPK) signalling and TNFα–NF-κB in Marie Ménard-fed rats. In the stools of the animals of this group we also measured a significant reduction of bacteria of the Bacteriodes fragilis group. In conclusion, the administration of Marie Ménard apples, rich in polyphenols and used at present only in the manufacturing of cider, ameliorates colon inflammation in transgenic rats developing spontaneous intestinal inflammation, suggesting the possible use of these and other apple varieties to control inflammation in IBD patients.

The development of technologies capable of early tumor detection is unquestionably demanded by physicians, as early diagnosis is key to achieve more efficient and less invasive treatments with improved outcomes. At the preclinical level, nanotechnology has already provided innovative solutions for tumor imaging and therapy, but it has failed to provide real early tumor diagnosis. In this work, an infrared nanothermometry‐based approach toward early melanoma detection, based on the changes produced in the thermal relaxation dynamics of tissues as the tumor develops, is introduced. In vivo experiments demonstrate that detection of incipient tumors from their very onset is possible through monitoring changes in their thermal relaxation dynamics using Ag2S infrared luminescent nanothermometers. For a total tumor development time of 14 days, luminescence nanothermometry allows tumor detection 6 days before its presence is evident by visual inspection. Simultaneous study of the tumoral vasculature reveals that the premature variation in the thermal relaxation dynamics is a consequence of the interplay between tumor angiogenesis and necrosis during the different tumor development stages.

The present paper deals with morphology of freshly dissected out live embryos of medicinally important plant, Basella alba Linn. The various part of the plant is used to cure dysentery, diarrhea, anemia, cancer etc. It is also has a known antioxidant, anti mutagenic, infertility, central nervous system depressant and anti inflammatory activity. The study of embryo at maturity in its native form is very significant, since its normal development is related to the viability of the seed. The young and mature embryo can be distinguished with the help of presence of chlorophyll. The viability of seeds is determined by the presence of green embryo at maturity, while nonviable seeds contains white coloured embryo. The embryo is coiled twice at young stage, while at maturity it is coiled one and half times, with the growing tip situated at the centre and the radicle on the outer side. Considering the medicinal value of B. alba, study the reproductive biology with respect to structure of matu...

Copper (Cu)-doped calcium silicate nanoparticles were synthesized by a wet precipitation method as economical bone fracture filler. The aim was to improve the overall physicochemical properties, bioactivity, and biological performance of the bone fracture filler prepared herein. The synthesized nanoparticles were evaluated using X-ray diffraction
(XRD), Fourier transform infrared spectroscopy (FT-IR), and
transmission electron microscopy (TEM). The bioactivity of the prepared nanoparticles was investigated after immersion in simulated body fluid (SBF) by means of inductively coupled plasma (ICP), SEM coupled with energy dispersive Xrays (EDX), and FTIR. The size and bioactivity of the prepared nanoparticles after 15 days of immersion in SBF was dependent on the Cu concentrations. The fracture healing ability of
the fabricated nanoparticles on adult aged male Wistar rats
was enhanced by the presence of copper. All the obtained
results are of high relevance for fabricating improved Cudoped
calcium silicate nanoparticles (50 nm) as low cost bone fracture filler. In addition, the in vivo study presented complete healing of the tibiae bone with normal architecture of bone tissue specifically calcium silicate nanoparticles doped with 3% and 5% Cu. Hence, the presence of copper is a promising tactic for improving the biological properties of
calcium silicate. Therefore, the designed nanoparticles have
huge potential for the treatment of bone fractures

Objective: Implantable neural electrode devices are important tools for neuroscience research and have an increasing range of clinical applications. However, the intricacies of the biological response after implantation, and their ultimate impact on recording performance, remain challenging to elucidate. Establishing a relationship between the neurobiology and chronic recording performance is confounded by technical challenges related to traditional electrophysiological, material, and histological limitations. This can greatly impact the interpretations of results pertaining to device performance and tissue health surrounding the implant. &#13; Approach: In this work, electrophysiological activity and immunohistological analysis are compared after controlling for motion artifacts, quiescent neuronal activity, and material failure of devices in order to better understand the relationship between histology and electrophysiological outcomes. &#13; Results: Even after carefully accounting for these factors, the presence of viable neurons and lack or glial scarring does not convey single unit recording performance. &#13; Significance: To better understand the biological factors influencing neural activity, detailed cellular and molecular tissue responses were examined. Decreases in neural activity and blood oxygenation in the tissue surrounding the implant, shift in expression levels of vesicular transporter proteins and ion channels, axon and myelin injury, and interrupted blood flow in nearby capillaries can impact neural activity around implanted neural interfaces. Combined, these tissue changes highlight the need for more comprehensive, basic science research to elucidate the relationship between biology and chronic electrophysiology performance in order to advance neural technologies.

Ethyl acetate extracts of Punica granatum were investigated for antitrypanosomal activities in Balb albino mice infected with Trypanosoma brucei brucei. The plant extracts at 100mg/kg, 200kg/kg and 300mg/kg body weight were administered intraperitoneally twice daily for 7 days to determine their trypanocidal activity. Ethyl acetate extract had a lethal dose (LD50) of 900 mg/kg. Treatment commenced on day 8 post infection in all groups except the negatve control group. Parasitaemia and disappearance of clinical signs were used as indices for monitoring the efficacy of the extracts using the rapid
matching method. Mean survival time showed that mice treated with diminazene aceturate survived for 15 days. The group which was administered 200mg/kg ethyl acetate leaf extract of Punica granatum had the highest mean survival time(9 ± 2.94 days) than those that received 100mg/kg((8 ± 0.82 days) and 300mg/kg(6.75 ± 0.5 days). Packed cell volume of the mice
changed significantly (p<0.05) in the group which received 200mg/kg ethyl acetate extract of Punica granatum. This dose has promising trypanocidal activity in vivo against Trypanosoma brucei brucei.

A new generation of bio-derived ceramics can be developed as a base material for medical implants. Specific plant species are used as templates on which innovative transformation processes can modify the chemical composition maintaining the original biostructure. Building on the outstanding mechanical properties of the starting lignocellulosic templates, it is possible to develop lightweight and high-strength scaffolds for bone substitution. In vitro and in vivo experiments demonstrate the excellent biocompatibility of this new silicon carbide material (bioSiC) and how it gets colonized by the hosting bone tissue because of its unique interconnected hierarchic porosity, which opens the door to new biomedical applications. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009

CRISPR-Cas9 has revolutionized the way we think about biology and has given scientists a tool to directly rewrite the code of life. Building on a mechanism found in some prokaryotes, scientists have created RNA guided " scissors " which can introduce highly accurate double stranded breaks in DNA. The genome can then be manipulated with surgical accuracy to generate mutants which model human diseases, or to correct dysfunctional genes in germ line cells, zygotes and post-natal somatic cells, without deleterious effects on cell health or function common to other past genomic editing technologies e.g. ZFNs or TALENS. CRISPR promises to usher in a new age of medicine and life sciences that –with caution and sound ethics, may one day soon lead to the eradication of a host of illnesses or perhaps extend the human lifespan long beyond its current state.

A new generation of bio-derived ceramics can be developed as a base material for medical implants. Specific plant species are used as templates on which innovative transformation processes can modify the chemical composition maintaining the original biostructure. Building on the outstanding mechanical properties of the starting lignocellulosic templates, it is possible to develop lightweight and high-strength scaffolds for bone substitution. In vitro and in vivo experiments demonstrate the excellent biocompatibility of this new silicon carbide material (bioSiC) and how it gets colonized by the hosting bone tissue because of its unique interconnected hierarchic porosity, which opens the door to new biomedical applications.

Objectives: To design a device that allows the formation of in situ oral biofilm with similar characteristics to those from the dental plaque, overcoming the limitations of previous devices.
Study Design: The Intraoral Device of Overlaid Disk-holding Splints (IDODS) was designed and manufactured. To test its validity, five healthy adult volunteers wore them for two and four days allowing the biofilm to grow without any type of distortion. After each period, the thickness, vitality and structure of the formed biofilm were measured with a Confocal Laser Scanning Microscope (CLSM) in combination with a dual fluorescence solution. All volunteers
filled out a Likert-type questionnaire to evaluate the device.
Results: Mean bacterial vitality in the 2- and 4-day biofilms was 71% and 63%, respectively. Mean thicknesses were 21 μm and 28 μm, respectively. There was predominance in the open and heterogeneous structure whose complexity
was ascending as the biofilm matured. The results obtained from the questionnaire were 2/5 in the influence in aesthetics, 3.4/5 in comfort, and 5/5 in ease of maintaining oral hygiene and withdrawal from the oral cavity.
Conclusions: A biofilm with optimum characteristics was obtained by IDODS. Its use is associated with good aesthetic and comfort results and is absent of functional limitations, allowing optimal oral hygiene without altering the structure of the in situ oral biofilm.