A multiscale model of the role of microenvironmental factors in cell segregation and heterogeneity in breast cancer development.

We analyzed a quantitative multiscale model that describes the epigenetic dynamics during the growth and evolution of an avascular tumor. A gene regulatory network (GRN) formed by a set of ten genes that are believed to play an important role in breast cancer development was kinetically coupled to the microenvironmental agents: glucose, estrogens, and oxygen. The dynamics of spontaneous mutations was described by a Yule-Furry master equation whose solution represents the probability that a given cell in the tissue undergoes a certain number of mutations at a given time. We assumed that the mutation rate is modified by a spatial gradient of nutrients. The tumor mass was simulated by means of cellular automata supplemented with a set of reaction diffusion equations that described the transport of microenvironmental agents. By analyzing the epigenetic state space described by the GRN dynamics, we found three attractors that were identified with cellular epigenetic states: normal, precancer and cancer. For two-dimensional (2D) and three-dimensional (3D) tumors we calculated the spatial distribution of the following quantities: (i) number of mutations, (ii) mutation of each gene and, (iii) phenotypes. Using estrogen as the principal microenvironmental agent that regulates cell proliferation process, we obtained tumor shapes for different values of estrogen consumption and supply rates. It was found that he majority of mutations occurred in cells that were located close to the 2D tumor perimeter or close to the 3D tumor surface. Also, it was found that the occurrence of different phenotypes in the tumor are controlled by estrogen concentration levels since they can change the individual cell threshold and gene expression levels. All results were consistently observed for 2D and 3D tumors.

An in vitro rumen incubation method to study exsheathment kinetics of Haemonchus contortus third-stage infective larvae.

This study developed and evaluated an in vitro rumen incubation (IVRI) method to describe the exsheathment kinetics of Haemonchus contortus third-stage infective larvae (L3) in ruminal liquor (RL). The specific objectives were (i) to standardize the IVRI method to facilitate the contact between L3 and RL as well as the larval recovery, and (ii) to apply the IVRI method to describe the exsheathment kinetics of H. contortus and to select the best fitting nonlinear model. Incubation devices containing H. contortus larvae were incubated according to the IVRI technique in cattle RL or PBS. The incubation conditions included RL mixed with a nitrogen-rich media, maintained at 39 °C, with pH = 7.0, vented with CO2 and manual agitation. The larvae were recovered after 0, 1, 3, 6, 9, 12, and 24 h. The exsheathed and ensheathed larvae were counted to estimate the exsheathment (%) in RL or PBS. Exsheathment in RL was analyzed with nonlinear regression models: Exponential, Gompertz, Logistic, Log-Logistic, and Weibull. The models' fit was compared to select the one that best described the exsheathment kinetics. The exsheathment in RL reached 6.52%, 20.65%, 58.22%, 69.24%, 73.08%, and 77.20% in 1, 3, 6, 9, 12, and 24 h, respectively. Although the Gompertz, Weibull, and Logistic models were adequate to describe the observed exsheathment, the Log-Logistic model had the best fit. The IVRI method using bovine RL represents a suitable tool for the study of the in vitro exsheathment kinetics of H. contortus L3.

Thermal Conductivity of Metastable Ionic Liquid [C2mim][CH3SO3].

Ionic liquids have been suggested as new engineering fluids, namely in the area of heat transfer, as alternatives to current biphenyl and diphenyl oxide, alkylated aromatics and dimethyl polysiloxane oils, which degrade above 200 °C and pose some environmental problems. Recently, we have proposed 1-ethyl-3-methylimidazolium methanesulfonate, [C2mim][CH3SO3], as a new heat transfer fluid, because of its thermophysical and toxicological properties. However, there are some interesting points raised in this work, namely the possibility of the existence of liquid metastability below the melting point (303 K) or second order-disorder transitions (l-type) before reaching the calorimetric freezing point. This paper analyses in more detail this zone of the phase diagram of the pure fluid, by reporting accurate thermal-conductivity measurements between 278 and 355 K with an estimated uncertainty of 2% at a 95% confidence level. A new value of the melting temperature is also reported, Tmelt = 307.8 ± 1 K. Results obtained support liquid metastability behaviour in the solid-phase region and permit the use of this ionic liquid at a heat transfer fluid at temperatures below its melting point. Thermal conductivity models based on Bridgman theory and estimation formulas were also used in this work, failing to predict the experimental data within its uncertainty.

MnTBAP Inhibits Bleomycin-Induced Pulmonary Fibrosis by Regulating VEGF and Wnt Signaling.

Cellular oxidative stress is implicated not only in lung injury but also in contributing to the development of pulmonary fibrosis. We demonstrate that a cell-permeable superoxide dismutase (SOD) mimetic and peroxynitrite scavenger, manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (MnTBAP) significantly inhibited bleomycin-induced fibrogenic effects both in vitro and in vivo. Further investigation into the underlying mechanisms revealed that MnTBAP targets canonical Wnt and non-canonical Wnt/Ca2+ signaling pathways, both of which were upregulated by bleomycin treatment. The effect of MnTBAP on canonical Wnt signaling was significant in vivo but inconclusive in vitro and the non-canonical Wnt/Ca2+ signaling pathway was observed to be the predominant pathway regulated by MnTBAP in bleomycin-induced pulmonary fibrosis. Furthermore, we show that the inhibitory effects of MnTBAP involve regulation of VEGF which is upstream of the Wnt signaling pathway. Overall, the data show that the superoxide scavenger MnTBAP attenuates bleomycin-induced pulmonary fibrosis by targeting VEGF and Wnt signaling pathways. J. Cell. Physiol. 232: 506-516, 2017. © 2016 Wiley Periodicals, Inc.

Med12 regulates ovarian steroidogenesis, uterine development and maternal effects in the mammalian egg.

The transcriptional factor MED12 is part of the essential mediator transcriptional complex that acts as a transcriptional coactivator in all eukaryotes. Missense gain-of-function mutations in human MED12 are associated with uterine leiomyomas, yet the role of MED12 deficiency in tumorigenesis and reproductive biology has not been fully explored. We generated a Med12 reproductive conditional knockout mouse model to evaluate its role in uterine mesenchyme, granulosa cells, and oocytes. Mice heterozygous for Med12 deficiency in granulosa cells and uterus (Med12fl/+ Amhr2-Cre) were subfertile, while mice homozygous for Med12 deficiency in granulosa cells and uterus (Med12fl/fl Amhr2-Cre) were infertile. Morphological and histological analysis of the Med12fl/fl Amhr2-Cre reproductive tract revealed atrophic uteri and hyperchromatic granulosa cells with disrupted expression of Lhcgr, Esr1, and Esr2. Med12fl/fl Amhr2-Cre mice estrous cycle was disrupted, and serum analysis showed blunted rise in estradiol in response to pregnant mare serum gonadotropin. Uterine atrophy was partially rescued by exogenous steroid supplementation with dysregulation of Notch1 and Smo expression in steroid supplemented Med12fl/fl Amhr2-Cre uteri, indicating intrinsic uterine defects. Oocyte-specific ablation of Med12 caused infertility without disrupting normal folliculogenesis and ovulation, consistent with maternal effects of Med12 in early embryo development. These results show the critical importance of Med12 in reproductive tract development and that Med12 loss of function does not cause tumorigenesis in reproductive tissues.

Molecular interactions and thermal transport in ionic liquids with carbon nanomaterials.

We used molecular dynamics simulation to study the effect of suspended carbon nanomaterials, nanotubes and graphene sheets, on the thermal conductivity of ionic liquids, an issue related to understanding the properties of nanofluids. One important aspect that we developed is an atomistic model of the interactions between the organic ions and carbon nanomaterials, so we did not rely on existing force fields for small organic molecules or assume simple combining rules to describe the interactions at the liquid/material interface. Instead, we used quantum calculations with a density functional suitable for non-covalent interactions to parameterize an interaction model, including van der Waals terms and also atomic partial charges on the materials. We fitted a n-m interaction potential function with n values of 9 or 10 and m values between 5 and 8, so a 12-6 Lennard-Jones function would not fit the quantum calculations. For the atoms of ionic liquids and carbon nanomaterials interacting among themselves, we adopted existing models from the literature. We studied the imidazolium ionic liquids [C4C1im][SCN], [C4C1im][N(CN)2], [C4C1im][C(CN)3] and [C4C1im][(CF3SO2)2N]. Attraction is stronger for cations (than for anions) above and below the π-system of the nanomaterials, whereas anions show stronger attraction for the hydrogenated edges. The ordering of ions around and inside (7,7) and (10,10) single-walled nanotubes, and near a stack of graphene sheets, was analysed in terms of density distribution functions. We verified that anions are found, as well as cations, in the first interfacial layer interacting with the materials, which is surprising given the interaction potential surfaces. The thermal conductivity of the ionic liquids and of composite systems containing one nanotube or one graphene stack in suspension was calculated using non-equilibrium molecular dynamics. Thermal conductivity was calculated along the axis of the nanotube and across the planes of graphene, in order to see the anisotropy. In the composite systems containing the nanotube, there is an enhancement of the overall thermal conductivity, with calculated values comparing well with experiments on nanotube suspensions, namely in terms of the order of the different ionic liquids. In the systems containing the graphene stack, the interfacial region of the ionic liquid near the surface of the material has an enhanced thermal conductivity with respect to the bulk liquid, but no significant discontinuity in the temperature profiles were observed. This is important information for models of thermal conduction in nanofluids.

Nitric oxide mediates bleomycin-induced angiogenesis and pulmonary fibrosis via regulation of VEGF.

Pulmonary fibrosis is a progressive lung disease hallmarked by increased fibroblast proliferation, amplified levels of extracellular matrix deposition and increased angiogenesis. Although dysregulation of angiogenic mediators has been implicated in pulmonary fibrosis, the specific rate-limiting angiogenic markers involved and their role in the progression of pulmonary fibrosis remains unclear. We demonstrate that bleomycin treatment induces angiogenesis, and inhibition of the central angiogenic mediator VEGF using anti-VEGF antibody CBO-P11 significantly attenuates bleomycin-induced pulmonary fibrosis in vivo. Bleomycin-induced nitric oxide (NO) was observed to be the key upstream regulator of VEGF via the PI3k/Akt pathway. VEGF regulated other important angiogenic proteins including PAI-1 and IL-8 in response to bleomycin exposure. Inhibition of NO and VEGF activity significantly mitigated bleomycin-induced angiogenic and fibrogenic responses. NO and VEGF are key mediators of bleomycin-induced pulmonary fibrosis, and could serve as important targets against this debilitating disease. Overall, our data suggests an important role for angiogenic mediators in the pathogenesis of bleomycin-induced pulmonary fibrosis.

Nicotine exposure during differentiation causes inhibition of N-myc expression.

BACKGROUND: The ability of chemicals to disrupt neonatal development can be studied using embryonic stem cells (ESC). One such chemical is nicotine. Prenatal nicotine exposure is known to affect postnatal lung function, although the mechanisms by which it has this effect are not clear. Since fibroblasts are a critical component of the developing lung, providing structure and secreting paracrine factors that are essential to epithelialization, this study focuses on the differentiation of ESC into fibroblasts using a directed differentiation protocol. METHODS: Fibroblasts obtained from non-human primate ESC (nhpESC) differentiation were analyzed by immunohistochemistry, immunostaining, Affymetrix gene expression array, qPCR, and immunoblotting. RESULTS: Results of these analyses demonstrated that although nhpESCs differentiate into fibroblasts in the presence of nicotine and appear normal by some measures, including H&E and SMA staining, they have an altered gene expression profile. Network analysis of expression changes demonstrated an over-representation of cell-cycle related genes with downregulation of N-myc as a central regulator in the pathway. Further investigation demonstrated that cells differentiated in the presence of nicotine had decreased N-myc mRNA and protein expression and longer doubling times, a biological effect consistent with downregulation of N-myc. CONCLUSIONS: This study is the first to use primate ESC to demonstrate that nicotine can affect cellular differentiation from pluripotency into fibroblasts, and in particular, mediate N-myc expression in differentiating ESCs. Given the crucial role of fibroblasts throughout the body, this has important implications for the effect of cigarette smoke exposure on human development not only in the lung, but in organogenesis in general.

Generation of two isogenic sickle cell disease induced pluripotent stem cell lines from testicular fibroblasts.

Curative bone marrow transplantation (BMT) therapies for sickle cell disease (SCD) can cause infertility. The Fertility Preservation Program (FPP) in Pittsburgh cryopreserves testicular tissues for SCD patients prior to BMT in anticipation that those tissues can be thawed in the future and matured to produce sperm. Here, we generated and validated two isogenic patient-derived induced pluripotent stem cell (iPSC) lines from testicular biopsy fibroblasts of a 12-year-old SCD patient.

Composition of Slow-Growing Male Chicken's Meat and Bone Quality as Affected by Dietary Moringa oleifera Lam. Meal.

This study investigated the impact of Moringa oleifera Lam. meal (MOM) on meat nutritional properties and bone quality of slow-growing layer-type male chickens raised in semi-intensive conditions. A total of 198, 72-d-old Dominant Blue D 107 male chickens, with an average weight of 1093 ± 15.2 g, were randomly assigned to three dietary treatments supplemented with 0, 3, and 6% of MOM that corresponded to T1, T2, and T3, respectively. Each treatment, consisting of six replicated floor pens of 11 birds, had access to the outdoors for 49 days. The results showed that breast muscle ash percentage was significantly greater (P ≤ 0.05) in T2 in comparison to the T1 group. Meat dry matter, protein, and fat content were not influenced by the treatments (P > 0.05). Regardless of the treatments, oleic acid (C18:1N9C) was numerically more abundant in the breast than in the leg muscle. Alternatively, femoral and tibial lengths were shorter (P ≤ 0.05) in birds fed 3% MOM than the two other groups. Moreover, birds fed with MOM had greater tibial diameter (P ≤ 0.05) than those that were fed without MOM. In addition, bone ash content and phosphorous amount were significantly higher (P ≤ 0.05) in birds fed 6% MOM compared to those fed without MOM. The data of this study indicate that up to 6% of MOM may be added to the diet of slow-growing layer-type male chickens raised with outdoor access under tropical conditions to improve bone quality traits.

High-Performance Ionanofluids from Subzipped Carbon Nanotube Networks.

Investments in the transfer and storage of thermal energy along with renewable energy sources strengthen health and economic infrastructure. These factors intensify energy diversification and the more rapid post-COVID recovery of economies. Ionanofluids (INFs) composed of long multiwalled carbon nanotubes (MWCNTs) rich in sp2-hybridized atoms and ionic liquids (ILs) display excellent thermal conductivity enhancement with respect to the pure IL, high thermal stability, and attractive rheology. However, the influence of the morphology, physicochemistry of nanoparticles and the IL-nanostructure interactions on the mechanism of heat transfer and rheological properties of INFs remain unidentified. Here, we show that intertube nanolayer coalescence, supported by 1D geometry assembly, leads to the subzipping of MWCNT bundles and formation of thermal bridges toward 3D networks in the whole INF volume. We identified stable networks of straight and bent MWCNTs separated by a layer of ions at the junctions. We found that the interactions between the ultrasonication-induced breaking nanotubes and the cations were covalent in nature. Furthermore, we found that the ionic layer imposed by close MWCNT surfaces favored enrichment of the cis conformer of the bis(trifluoromethylsulfonyl)imide anion. Our results demonstrate how the molecular perfection of the MWCNT structure with its supramolecular arrangement affects the extraordinary thermal conductivity enhancement of INFs. Thus, we gave the realistic description of the interactions at the IL-CNT interface with its (super)structure and chemistry as well as the molecular structure of the continuous phase. We anticipate our results to be a starting point for more complex studies on the supramolecular zipping mechanism. For example, ionically functionalized MWCNTs toward polyionic systems─of projected and controlled nanolayers─could enable the design of even more efficient heat-transfer fluids and miniaturization of flexible electronics.

Acinetobacter baumannii is not associated with osteomyelitis in a rat model: a pilot study.

BACKGROUND: Multidrug resistant Acinetobacter baumannii (MDR AB) with and without Staphylococcus aureus (SA) is a commonly isolated organism in infected segmental bone defects in combat-related trauma in Iraq and Afghanistan. Although MDR AB in visceral infections is a therapeutic challenge, control of infection appears more common for combat-related osteomyelitis. QUESTIONS/PURPOSES: Using a rat model, we explored the virulence of MDR AB in segmental bone defects alone and in combination with SA. METHODS: Segmental defects in 60 rat femurs were created, stabilized, and inoculated with MDR AB alone and 60 with MDR AB and SA. We performed qualitative and quantitative bacteriology and radiographic assessments at 2, 4, and 8 weeks for MDR AB and at 1, 2, and 3 weeks for MDR AB and SA. RESULTS: Quantitative bacteriology revealed a 3- to 5-log decrease in MDR AB from the initial inoculum. After polymicrobial inoculation, only 10 of 60 animals had positive cultures for MDR AB, whereas 59 of 60 animals had positive cultures for SA. Recovered SA were 2 to 5 log greater than the initial inoculum, while there again was a 3- to 5-log decrease in MDR AB. MDR AB alone did not cause bony lysis, but there was radiographic evidence of new bone formation in 67% of the segmental defects. Osteolysis was noted with MDR AB and SA. CONCLUSIONS: MDR AB did not appear to cause or contribute to clinically apparent osteomyelitis in this pilot study. CLINICAL RELEVANCE: Resolution of infections in combat-related segmental bone defects inoculated with MDR AB may be attributable to low virulence. Additional studies are needed to confirm low virulence and bone formation with MDR AB.

Metabolizable energy balance in hair sheep lambs artificially infected with Haemonchus contortus.

In sheep, infection with Haemonchus contortus may increase the need for energy, and this demand may vary according to the infection level. In this study, the energy intake, digestibility, and energy retention of lambs artificially infected with different levels of H. contortus were estimated. A total of 24 hair sheep lambs reared parasite-free were experimentally infected with H. contortus at one of three infection levels: non-infected (n = 6); infected with 300 infective larvae (L3) of H. contortus/kg body weight (BW) (n = 9); and infected with 500 H. contortus L3/kg BW (n = 9). The lambs were fed for an individual weight gain of 100 g/day, and intake of organic matter (OMI) and gross energy (GEI), digestible (DEI) and metabolizable energy (MEI) were measured weekly. The digestibility of organic matter (OMD) and GE (GED) and the metabolizable energy (ME) balance adjusted to zero nitrogen balance (MEadj) were measured for each lamb during the prepatent and patent periods of infection. From day 21 post-infection (PI), the individual eggs per gram (EPG) of feces and the total number of eggs in feces (TEF) were estimated weekly. After humane slaughter on day 42 PI, the worm burden (WB) was determined. Correlation and regression analyses were performed to estimate the relationships between the parasitological variables (L3, EPG, TEF and WB) and the response variables (OMI, GEI, DEI, MEI, OMD, GED, MEadj). During the prepatent period, there were no significant relationships of L3 with the response variables (OMI, GEI, DEI, MEI, OMD, GED, ME, MEadj). Similarly, during the patent period, no relationship was evident between infection (EPG, TEF or WB) and OMI, GEI, DEI, GED, OMD, ME or MEadj. Thus, the gradient of H. contortus infection examined in the present study did not influence energy balance in hair sheep lambs, and infection did not impose any detectable energy cost. Further studies are needed to fully assess the impact of H. contortus infection on energy metabolism in hair sheep.

Analysis of dietary patterns and nutritional adequacy in lactating women: a multicentre European cohort (ATLAS study).

Eating habits of lactating women can influence the nutrient composition of human milk, which in turn influences nutrient intake of breastfed infants. The aim of the present study was to identify food patterns and nutritional adequacy among lactating women in Europe. Data from a multicentre European longitudinal cohort (ATLAS study) were analysed to identify dietary patterns using cluster analysis. Dietary information from 180 lactating women was obtained using 3-d food diaries over the first 4 months of lactation. Four dietary patterns were identified: 'vege-oils', 'fish-poultry', 'confectionery-salads' and 'mixed dishes'. Nutrition adequacy was not significantly different between clusters, but the 'vege-oils' cluster tended to yield the highest nutrition adequacy measured by Mean Adequacy Ratio. Compared with European dietary reference values (DRVs) for lactating women, women in all clusters had inadequate intakes of energy, pantothenic acid, folate, vitamin C, vitamin A, vitamin D, zinc, iodine, potassium and linoleic acid. Adequate intake for fibre and α-linolenic acid was only achieved in the 'vege-oils' cluster. Overall, fat intake was above DRVs. The present study showed that various dietary patterns do not adequately supply all nutrients, indicating a need to promote overall healthy dietary habits for European lactating women.

Prevalence, estimated HIV-1 incidence and viral diversity among people seeking voluntary counseling and testing services in Rio de Janeiro, Brazil.

BACKGROUND: BED-EIA HIV-1 Incidence Test (BED-CEIA) has been described as a tool to discriminate recent (RS) from long-term (LTS) seroconversion of HIV-1 infection, contributing to a better understanding of the dynamics of the HIV/AIDS epidemic over time. This study determined the prevalence, estimated incidence and HIV-1 subtype infection among individuals seeking testing in Voluntary Counseling and Testing centers (VCTs) from Rio de Janeiro, Brazil. METHODS: Demographics and behavioral data were obtained from 434 individuals, diagnosed as HIV-positive among 9,008 volunteers screened from November 2004 to October 2005 in three VCTs located in the Rio de Janeiro Metropolitan area, Brazil. BED-CEIA protocol was performed to identify RS. DNA samples from RS and a subset of LTS (under a proportion of 1:2) were selected for gp120 C2-V3 and pol (protease and reverse transcriptase) regions genomic sequencing. RESULTS: Overall HIV-1 prevalence was 4.8%. Sixty-one of 434 seropositive individuals were classified as RS, corresponding to an incidence rate of 1.68%/year (95%CI 1.26% -2.10%). Estimated incidence between Men Who Have Sex with Men (MSM) was 11 times higher than among heterosexual men and 55% of the new cases were identified in volunteers aged 25-40 years. A similar distribution of different HIV-1 subtypes was found among RS and LTS. CONCLUSIONS: Our data suggest that prevention for MSM remains a challenge and efforts focusing on prevention targeting this population should be prioritized. No significant changes in HIV-1 subtypes were observed among the RS and LTS subgroups. One case of HIV-1 AUK (pol)/A (env) recombinant genome was detected for the first time in Brazil.

Evaluation of a lumbar intervertebral spacer with integrated screws as a stand-alone fixation device.

STUDY DESIGN: An in vitro cadaveric biomechanics study. OBJECTIVE: To evaluate the biomechanical contribution of the integration of screws into a polyether-ether-ketone (PEEK) spacer, and to compare the resulting construct to standard anterior lumbar interbody fusion constructs. SUMMARY OF BACKGROUND DATA: Pedicular fixation is often performed to increase the rigidity of anterior lumbar interbody fusion constructs but also increases the risk of morbidity. Integration of screws into a PEEK spacer (investigational device, ID) may increase construct rigidity and prevent migration without the drawbacks associated with supplementary fixation. METHODS: Twenty cadaveric motion segments were potted and tested under pure moments of + or - 7.5 Nm in flexion-extension, lateral bending, and axial torsion. Discectomies were performed and specimens were instrumented with the ID + or - screws, dual threaded cages, structural graft+anterior plate, and graft+pedicle screws. The ID+screws and threaded cage constructs were then pulled out at a constant rate. RESULTS: All constructs reduced the mean range of motion (ROM) below the intact level in flexion-extension and lateral bending (P<0.001) and for all devices except cages in torsion (P < or = 0.05). The median flexion-extension/bending/torsion ROM was 9.5/9.4/4.1 degrees for the intact segments, 6.1/5.1/1.8 degrees with the ID+screws, 4.9/5.2/2.4 degrees with threaded cages, 3.3/4.4/1.1 degrees with plates and 1.4/1.6/1.7 degrees with pedicle screws, respectively. The addition of the integrated screws decreased the mean ROM of the ID constructs by 0.9 degrees in flexion-extension, 1.8 degrees in bending, and 2.8 degrees in flexion-extension. The peak pullout force was 962 N for the ID and 337 N for threaded cages. CONCLUSIONS: The incorporation of screws into a PEEK interbody device was evaluated alongside traditional constructs in terms of construct rigidity and resistance to pullout. The greatest contributions of the integrated screws are in limiting torsional motion and in the containment of the interbody device. Pedicular fixation produces the most rigid constructs, but integrating screws into a PEEK spacer produces a stand-alone construct that is similar to threaded cages and provides more resistance to anterior displacement.

Host tissue response to floating microelectrode arrays chronically implanted in the feline spinal nerve.

OBJECTIVE: Neural interfacing technologies could significantly improve quality of life for people living with the loss of a limb. Both motor commands and sensory feedback must be considered; these complementary systems are segregated from one another in the spinal nerve. APPROACH: The dorsal root ganglion-ventral root (DRG-VR) complex was targeted chronically with floating microelectrode arrays designed to record from motor neuron axons in the VR or stimulate sensory neurons in the DRG. Hematoxylin and eosin and Nissl/Luxol fast blue staining were performed. Characterization of the tissue response in regions of interest and pixel-based image analyses were used to quantify MAC387 (monocytes/macrophages), NF200 (axons), S100 (Schwann cells), vimentin (fibroblasts, endothelial cells, astrocytes), and GLUT1 (glucose transport proteins) reactivity. Implanted roots were compared to non-implanted roots and differences between the VR and DRG examined. MAIN RESULTS: The tissue response associated with chronic array implantation in this peripheral location is similar to that observed in central nervous system locations. Markers of inflammation were increased in implanted roots relative to control roots with MAC387 positive cells distributed throughout the region corresponding to the device footprint. Significant decreases in neuronal density and myelination were observed in both the VR, which contains only neuronal axons, and the DRG, which contains both neuronal axons and cell bodies. Notably, decreases in NF200 in the VR were observed only at implant times less than ten weeks. Observations related to the blood-nerve barrier and tissue integrity suggest that tissue remodeling occurs, particularly in the VR. SIGNIFICANCE: This study was designed to assess the viability of the DRG-VR complex as a site for neural interfacing applications and suggests that continued efforts to mitigate the tissue response will be critical to achieve the overall goal of a long-term, reliable neural interface.

Ensiling Process in Commercial Bales of Horticultural By-products from Artichoke and Broccoli.

Wastes from artichoke and broccoli crops and cannery industries represent an environmental problem. A viable option to this problem is ensiling them for use as ruminants feed. The aim of this study was to characterise the ensiling process of broccoli and artichoke by-products and assess their suitability to be part of the ruminant diet, as well their minimum shelf life. Twenty-one commercial round bale silos (300 kg and 0.64 m3) of each by-product were made. Samples were analysed at days 0, 7, 15, 30, 60, and 200 to determine microbial populations, fermentation metabolites, nutritional components, and phytosanitary residues. Feedstuffs showed good suitability for ensiling, and stabilisation was achieved on day 30. The variables with the greatest significant differences among sampling times were microbial populations and fermentative components. There were no important dry matter losses, and some significant differences were observed in the nutritional composition, especially in crude protein and fibrous fractions, but they were not relevant for the loss of nutritional quality of silages. The phytosanitary residues determined on day 200 were below the maximum residue limits set by European legislation. So, ensiling these by-products in commercial round bale silos is a suitable and profitable technique that allows their preservation for a long time (200 days).

Composition, Mineral and Fatty Acid Profiles of Milk from Goats Fed with Different Proportions of Broccoli and Artichoke Plant By-Products.

In the Mediterranean region, artichoke and broccoli are major crops with a high amount of by-products that can be used as alternative feedstuffs for ruminants, lowering feed costs and enhancing milk sustainability while reducing the environmental impact of dairy production. However, nutritional quality of milk needs to be assured under these production conditions and an optimal inclusion ratio of silages should be determined. This work aimed to evaluate the effect of three inclusion levels (25%, 40%, and 60%) of these silages (artichoke plant, AP, and broccoli by-product, BB) in goat diets on milk yield, composition, and mineral and fatty profiles. Treatments with 60% inclusion of AP and BB presented the lowest milk yield. No differences were found on the milk mineral profile. Inclusion of AP in the animals' diet improved the milk lipid profile from the point of view of human health (AI, TI) compared to BB due to a lower saturated fatty acid content (C12:0, C14:0, and C16:0) and a higher concentration of polyunsaturated fatty acids (PUFA), especially vaccenic acid (C18:1 trans11) and rumenic acid (CLA cis9, trans11), without any differences with the control treatment.

Immune activation and antibody responses in non-progressing elite controller individuals infected with HIV-1.

An extremely rare subset of patients infected with HIV-1 designated as "non-progressing elite controllers" appears to be able to maintain stable CD4(+) T-cell counts and a median plasma viremia below the detection limit of current ultrasensitive assays (<50-80 copies/ml of plasma) for >10 years in the absence of antiretroviral therapy. Lymphocyte subsets (CD4(+), CD8(+)), immune activation markers (HLA-DR(+), CD38(+), Beta-2-microglobulin), and HIV-specific antibody responses were longitudinally examined in four non-progressing elite controllers over more than 5 years. Two control groups of seronegative healthy individuals and untreated patients infected with HIV-1 presenting detectable viremia were also included. None of the non-progressing elite controllers displayed the high T-cell activation levels generally seen in the seropositive individuals, keeping them within the normal range. Three non-progressing elite controllers showed no significant immune system abnormalities when compared to seronegative individuals, displaying a low proportion of HIV-1-specific binding antibodies and low avidity index, similar to those observed for individuals infected recently with HIV-1. One non-progressing elite controller exhibited CD8(+) T-cell counts and beta2-M levels above normal ranges and developed a low but "mature" (high-avidity) HIV-1-specific antibody response. Thus, the non-progressing elite controllers are able to maintain normal T-cell activation levels, which may contribute to prevent, or greatly reduce, the damage of the immune system typically induced by the HIV-1 over time. They are, however, immunologically heterogeneous and very low levels of antigen exposure seem to occur in these patients, sufficient for sustaining a low, but detectable, HIV-1-specific immunity.