Bovine Brain Research Articles

Hyperinvasiveness of Listeria monocytogenes sequence type 1 is independent of lineage I-specific genes encoding internalin-like proteins.

Listeriosis is a severe disease caused by the opportunistic bacterial pathogen Listeria monocytogenes (L. monocytogenes). Previous studies indicate that of the four phylogenetical lineages known, lineage I strains are significantly more prevalent in clinical infections than in the environment. Among lineage 1, sequence type (ST1) belongs to the most frequent genotypes in clinical infections and behaves hyperinvasive in experimental in vitro infections compared to lineage II strains suggesting that yet uncharacterized virulence genes contribute to high virulence of certain lineage I strains. This study investigated the effect of four specific lineage I genes encoding surface proteins with internalin‐like structures on cellular infection. CNS derived cell lines (fetal bovine brain cells, human microglia cells) and non‐CNS derived cell lines (bovine macrophage cells, human adenocarcinoma cells) that represent the various target cells of L. monocytogenes were infected with the parental ST1 strain and deletion mutants of the four genes. Despite their association with lineage I, deletion of the four genes investigated did not dampen the hyperinvasiveness of the ST1 strain. Similarly, these genes did not contribute to the intracellular survival and intercellular spread of L. monocytogenes ST1, indicating that these genes may have other functions, either during the infection process or outside the host.

Development of Computed Tomography Head and Body Phantom for Organ Dosimetry

Introduction: Quality assurance in Computed tomography (CT) centers in developing countries are largely hindered by the unavailability of CT phantoms. The development of a local CT phantom for the measurement of organ radiation absorbed dose is therefore requisite. Material and Methods: Local CT phantoms were designed to meet the standard criteria of 32 cm diameter for body, 16 cm diameter for head, and 14 cm in length respectively. The outer plastic shell was made using poly (methyl methacrylate [PMMA]) sheet. The developed CT phantoms were validated against a standard phantom. Radiation absorbed dose was determined by scanning the setup with the same protocol used for the standard phantom. The local phantoms were then verified for organ radiation absorbed dose measurement using bovine tissues. The set up was CT-scanned, and Hounsfield units (HU) for bovine tissues were obtained. Results: There was no significant difference between the local and standard head phantoms (P=0.060). Similarly, no difference was noted between the local and standard body phantoms (P=0.795). The percentage difference in volume CT dose index (CTDIvol) between the body (local and standard) phantoms was higher than that for the head phantoms. There were no significant differences in HU between bovine and human brain, liver, kidney and lung tissues (P=0.938). Conclusion: The local phantoms showed good agreement with the standard ones. The developed phantoms can be used for CT organ radiation absorbed dose measurement in radiology departments in Nigeria.

Easy Determination of Radiation Absorption in Brain Tissue from Mobile Phones Using Finite Element Method

Brain tissue plays a significant role in both cognitive and psychomotor behavior of humans. However, their interaction with radiation emanating from hand held mobile devices is still not fully understood. This research was aimed at investigating radiation absorption in brain tissue. Bovine brain tissues ranging from lesser than 1 year to greater than 10 years of age were bought from a specialty store (Sigma-Aldrich). The tissues were used within 72 h of extraction for ex vivo brain experiments. The brain tissue was stored at 6°C and then 16°C for 24 h in the MRI room to reach thermal equilibrium before any experiments were undertaken. The averages for the dielectric constant were measured from 1 - 4 GHz using open ended coaxial probe (OECP) (85,070E; Agilent Technologies). The results obtained for the dielectric properties were then used as raw data in the numerical computation and simulation of the radiation absorption by the brain tissues for both adolescent and adults bovine brain tissue using finite element method (FEM). The measured dielectric constants varied for the different brain tissue from 54.39 to 39.29. Analysis showed that adolescents tissue absorbed more radiation than adults from mobile phoneradiation which is due to the higher dielectric property of adolescent brain tissue. The results obtained can be applied to human brain tissue since bovine shares the same compositional properties with humans.

Myelin basic protein charge isomers change macrophage polarization.

PurposeDuring a neuronal injury, a variety of immune cells infiltrate into the local microenvironment at the demyelination site. After the destruction of the intact myelin sheath, its major constituent myelin basic protein (MBP) dissociates from the plasma membrane and acts as a free ligand on the infiltrated immune cells. MBP exhibits charge microheterogeneity as a result of post-translational modifications, but the effect of various isomers of MBP on the activity of macrophages is not known.Materials and methodsMBP was isolated and purified from bovine brain white matter. RAW 264.7 macrophages were cultured in DMEM supplemented with heat-inactivated fetal bovine serum. For evaluation of macrophage polarization following treatment of RAW 264.7 cells with MBP charge isomers, inducible nitric oxide synthase (iNOS) expression (M1 phenotype marker) and arginase-1 expression (M2 phenotype marker) were determined in cell lysates by ELISA. To assess Rac activity, G-LISA Rac Activation Assay system was used. The expression of receptor for advanced glycation end-products (RAGE) and high mobility group box 1 (HMGB1) protein were assayed by Western blot analysis.ResultsOur results have shown that minimally modified C1 component of MBP increases the expression of arginase-1 in cells, decreases the expression of iNOS, does not change the secretion of HMGB1 protein, but significantly elevates surface expression of RAGE, and in parallel, increases the activity of small GTPase Rac. On the other hand, highly modified deiminated isomer C8-MBP increases the secretion of HMGB1 protein but does not change the expression of arginase-1 or the content of RAGE.ConclusionThese data indicate that deiminated C8 isomer of MBP tends to polarize RAW macrophages into M1 phenotypes, whereas C1 enhances the activity of M2 phenotype markers.

COMP-19. WATER-CONTENT BASED ELECTRIC PROPERTY TOMOGRAPHY (wEPT) FOR MODELLING DELIVERY OF TUMOR TREATING FIELDS TO THE BRAIN

Understanding how Tumor Treating Fields (TTFields) distribute in the brain is important for optimizing delivery. The distribution depends on the electric properties (EPs) of the brain, which are heterogeneous. Therefore, there is a need for methods that map electric properties within tissue with high spatial resolution. Water content based EP tomography (wEPT) utilizes the ratio of two T1w to map tissue water content, and EPs. wEPT has been applied to map EPs at 128 MHz. This study examines the suitability of wEPT to map EPs in the brain at 100–1000 kHz (TTFields frequency range). MATERIALS AND A model connecting T1 images, Water content (WC) and EPs in the 100–1000 kHz range was created using tissue samples derived from bovine brains and aporcine CSF sample. For each sample, T1w images were acquired and WC and EPs measured. Curve fitting yielded models connecting Ir, WC and EPs. Next, wEPT was applied to map water content and EPs in tumor-bearing rat brains. EPs and WC were measured on 6 samples excised from each brain. The measured values were compared to the WC and EPs in the regions of the wEPT map corresponding to the sample. Water content estimated using wEPT agreed well with measurements on excised sample. A connection between EPs estimated with wEPT and the measured values was found. However, in some samples, especially samples excised from the tumors, large differences between wEPT-derived EP values and measurements existed. wEPT maps WC in healthy and tumor brain tissues However, the relationship between WC and EPs within this frequency range is not clear. Further work is required to refine this relationship enabling a robust non-invasive method for mapping electrical properties within the brain, and better strategies for optimizing TTFields treatment.

Gross and histopathological pitfalls found in the examination of 3,338 cattle brains submitted to the BSE surveillance program in Brazil

ABSTRACT: This study stems from the findings during the gross and histopathological exam of 3,338 cattle brains as part of the bovine spongiform encephalopathy (BSE) active surveillance program of the Brazilian Ministry of Agriculture, Livestock, and Supply from 2001 to 2005. The work was carried out in the Veterinary Pathology Laboratory of the Federal University of Santa Maria which at the time (2001-2007) was the national reference laboratory for the diagnosis of BSE and other transmissible spongiform encephalopathies. Both gross and histopathological aspects are described. Several gross aspects were annotated: anatomic normal structures not commonly recognized (non-lesions), lesions of no clinical significance, postmortem changes and artifacts; all these can amount to important pitfalls that distract the pathologist during the routine gross examination of the central nervous system (CNS). Accordingly, equivalent pitfalls were described in the histological examination. Non-lesions observed were the pineal body, embryo remnants such as the external germinal layer of the cerebellum, subependymal plates, and clusters of neuroblasts in the basal ganglia; or circumventricular structures such as area postrema, subcomisural organ, and melanosis in the leptomeninges and vessel walls. Lesions with little or no clinical importance included age-related changes as lipofuscin, hemosiderin, mineralization and hyalinization of vessel walls within the brain and meninges. Corpora amylacea and corpora arenacea were detected respectively in astrocyte processes and the pineal body. Cytoplasmic neuronal vacuolization was observed in the red nucleus and habenular nucleus. Sarcocystis sp. without a correspondent inflammatory reaction was rarely observed. Included within findings with no clinical manifestation were axonal spheroids and perivascular mononuclear cuffings. Changes in the CNS due to killing, sampling and fixation methods can obscure or distract from the more critical lesions. The ones related to the process of killing included hemorrhages caused in cattle destroyed by a captive bolt. Artifacts related to sampling and handling of CNS tissue consisted of inclusion of bone sand in the neural tissue from sawing the calvarium; dark neurons produced by excessive handling of the brain, and micro-organisms that contaminated the tissues during sampling or histological processing. Postmortem autolytic or putrefactive changes observed included vacuolar changes in the myelin sheath, clear halos surrounding neurons and oligodendrocytes, clusters of putrefaction bacilli within vessels or dispersed throughout the brain tissue associated or not to clear halos. One interesting, and somewhat frequent, postmortem autolytic change found in the bovine brain was the partial dissolution of the granule cell layer (GCL) of the cerebellum, also referred to as conglutination of the GCL or as the French denomination “état glace”. Due to the shortage of comprehensive publications in the subject, this review is intended to address the main pitfalls that can be observed in the brain of cattle hoping to help other pathologists avoiding misinterpret them.

Up-regulated expression of spherical body protein 2 truncated copy 11 in Babesia bovis is associated with reduced cytoadhesion to vascular endothelial cells

The factors involved in gain or loss of virulence in Babesia bovis are unknown. Spherical body protein 2 truncated copy 11 (sbp2t11) transcripts in B. bovis were recently reported to be a marker of attenuation for B. bovis strains. Increased cytoadhesion of B. bovis-infected red blood cells (iRBC) to vascular endothelial cells is associated with severe disease outcomes and an indicator of parasite virulence. Here, we created a stable B. bovis transfected line over-expressing sbp2t11 to determine whether up-regulation of sbp2t11 is associated with changes in cytoadhesion. This line was designated sbp2t11up and five B. bovis clonal lines were derived from the sbp2t11up line by limiting dilution for characterisation. We compared the ability of iRBCs from the sbp2t11up line and its five derivative clonal lines to adhere to bovine brain endothelial cells, using an in vitro cytoadhesion assay. The same lines were selected for in vitro cytoadhesion and the levels of sbp2t11 transcripts in each selected line were quantified. Our results demonstrate that up-regulation of sbp2t11 is accompanied by a statistically significant reduction in cytoadhesion. Confirmed up-regulation of sbp2t11 in B. bovis concomitant with the reduction of iRBC in vitro cytoadhesion to bovine brain endothelial cell is consistent with our previous finding that up-regulation of sbp2t11 is an attenuation marker in B. bovis and suggests the involvement of sbp2t11 transcription in B. bovis virulence.

The bovine anterior hypothalamus: Characterization of the vasopressin-oxytocin containing nucleus and changes in relation to sexual differentiation.

In an effort to systematically describe the neurochemical anatomy of the bovine anterior hypothalamus, we used a series of immunocytochemical markers such as acetylcholine esterase (AChE), arginine-vasopressin (AVP), calbindin (Calb), galanin (Gal), neuropeptide-Y (NPY), oxytocin (OXT), somatostatin (SST), and vasoactive intestinal peptide (VIP). We also investigated the potential sex difference present in the suprachiasmatic nucleus (SCN) and the vasopressin-oxytocin containing nucleus (VON) of six male and six female Bos taurus. Our study revealed that the cytochemical structure of the cattle anterior hypothalamus follows the blueprint of other mammals. The VON, which was never described before in cattle, showed a sex difference with a 33.7% smaller volume and 23.2% fewer magnocellular neurons (approximately 20-30 μm) in the male. The SCN also did show a sex difference in VIP neurons and volume with a 36.1% larger female nucleus with 28.1% more cells. Additionally, we included five heifers with freemartin syndrome as a new animal model relevant to sexual differentiation in the brain. This is, to the best of our knowledge, the first freemartin study in relation to the brain. Surprisingly, the SCN of freemartin heifers was 32.5% larger than its control male and female counterparts with 29% more VIP cells. Conversely, the freemartin VON had an intermediary size between male and female. To analyze our data, a classical statistical analysis and a novel multivariate and multi-aspect approach were applied. These findings shed new light on sexual dimorphism in the bovine brain and present this species with freemartins as a valuable animal model in neuroscience.

UM-HACC-2A: MYB-NFIB fusion-positive human adenoid cystic carcinoma cell line

ObjectivesLimited availability of validated human adenoid cystic carcinoma (ACC) cell lines has hindered the mechanistic understanding of the pathobiology of this malignancy and the development of effective therapies. The purpose of this work was to generate and characterize a human ACC cell line. Material and MethodsImmediately after surgery, a tumor fragment from a minor salivary gland from the tongue of a female Caucasian was minced, dissociated, and a single cell suspension was plated in fibronectin-coated flasks. A culture medium containing bovine brain extract and rhEGF was optimized for these cells. Whole exome sequencing was used to evaluate the presence of MYB-NFIB translocation. ResultsThe University of Michigan-Human Adenoid Cystic Carcinoma (UM-HACC)-2A cells showed continuous growth in monolayers for at least 180 in vitro passages while maintaining epithelial morphology. Short-tandem repeat (STR) profiling confirmed a 100% match to patient DNA. Whole exome sequencing revealed the presence of the MYB-NFIB fusion in UM-HACC-2A cells, which was confirmed by PCR analysis. Western blots revealed high expression of epithelial markers (e.g. E-cadherin, EGFR, pan-cytokeratin) and proteins associated with ACC (e.g. c-Myb, p63). Developmental therapeutic studies showed that UM-HACC-2A cells were resistant to cisplatin (IC50 = 44.7 µM) while more responsive to paclitaxel (IC50 = 0.0006 µM). In a pilot study, we observed that UM-HACC-2A cells survived orthotopic transplantation into the submandibular gland. Notably, one of the mice injected with UM-HACC-2A cells exhibited lung metastasis after 6 months. ConclusionUM-HACC-2A is a MYB-NFIB fusion-positive ACC cell line that is suitable for mechanistic and developmental therapeutics studies.

Peptide-functionalized and high drug loaded novel nanoparticles as dual-targeting drug delivery system for modulated and controlled release of paclitaxel to brain glioma

A dual-targeting drug delivery system for paclitaxel (PTX) was developed by functionalizing novel polyester-based nanoparticles (NPs) with peptides possessing special affinity for low-density lipoprotein receptor (LDLR), overcoming the limitations of the current chemotherapeutics, to transport drug from blood to brain, and then target glioma cells. Employing novel biodegradable block co-polymers (P and 2P), PTX loaded and peptide-functionalized nanoparticles were prepared by a modified nano-co-precipitation method, carried out in one step only without emulsifier, allowing to obtain spherical nanometric (<200 nm), monodisperse (PDI ∼ 0.1), Poly (Ethylene Glycol) (PEG)-coated and high PTX loaded NPs with a slow and controlled release rate for a prolonged period of time. Peptide functionalization, confirmed by fluorimetric assay and HPLC amino acids analysis, enhanced the cellular uptake of functionalized-PTX-NPs by human primary glioblastoma cell line (U-87 MG) and Bovine Brain Endothelial Cells (BBMVECs), compared with non-functionalized-PTX-NPs. To confirm dual-targeting effect, transendothelial transport experiments in an in vitro BBB model and in vitro anti-tumoral activity against U-87 MG revealed that peptide-functionalized-PTX-NPs significantly increased the transport ratio of PTX across the BBB along with an improved anti-proliferative efficiency. Pharmacokinetics and biodistribution studies in rats, carried out by in vivo experiments with 125I radiolabelled dual-targeting PTX-NPs, confirmed the stealthy behavior of NPs and indicated slightly lower levels of penetration into brain tissue in comparison with peptides known to be able to cross the BBB. These promising results suggested that the dual-targeting drug delivery system might have great potential for glioma therapy in clinical applications.

P04.29 Modelling delivery of Tumor Treating Fields (TTFields) to the brain using Water-based Electrical Properties Tomography

Tumor Treating Fields are anti-cancer treatment utilizing alternating electric fields in the intermediate frequency range. TTFields (@200 kHz) are approved for the treatment of glioblastoma multiforme. The effect of TTFields is intensity-dependent, suggesting it is important to maximize TTFields dose at the tumor. Hence, it is important to understand how TTFields distribute in the brain. The distribution of TTFields within the brain depends on the electric properties (EPs) of the brain, which are heterogeneous. Therefore, there is a need for methods that map electric properties within tissue with high spatial resolution. Water content based EP tomography (wEPT) is a method that utilizes the ratio of two T1w images with different relaxation times (TRs) to map EPs. wEPT has been applied to map EPs of healthy brain at 128 MHz. Here we attempted to adapt wEPT to map EPs in the 100–1000 kHz. An empirical model connecting T1 images, Water content (WC) and EPs in the 100–1000 kHz range was created using 32 tissue samples derived from three 3 juvenile bovine brains and 1 Porcine CSF sample. For each sample, T1w MRIs with TRs {700, 4000} ms were acquired and the image ratio (Ir) between the images calculated. EPs of samples were measured with a parallel plate setup. The dry and wet mass of the samples was measured for each sample, and the water content estimated from the difference between the two measurements. Curve fitting yielded empirical models connecting Ir, WC and EPs. Next, T1w MRIs of tumor-bearing rat brains in-vivo were acquired, and the empirical curves described above used to map WC and EP within the rat brains. In addition, EPs and WC were measured on 6 samples excised from each imaged brain. The measured values were compared to the median WC and EPs in the voxels of the wEPT map corresponding to the sample. Anatomical structures and the tumor were clearly visible in wEPT maps of the rat brains. WC estimated using wEPT agreed well with measurements on excised sample. The data showed a connection between EPs estimated with wEPT and the measured values. However, in some samples large differences between wEPT-derived EP values and measurements were found. In particular, differences between tumor conductivity and the conductivity of grey and white matter estimated using wEPT was significantly higher than the difference in conductivities measured within the excised samples. wEPT maps WC in healthy and tumor brain tissues and provides information on local electrical properties at frequencies of 100–1000 kHz. Further investigation is needed to clarify the relationship between WC and EP within this frequency range. Clarifying this relationship would enable a robust non-invasive method for tissue electrical properties within the brain, ultimately leading to a better understanding of how TTFields distribute within the brain and influence disease progression.

Mechanism of Galactocerebroside Adsorption on Silicalite-1

The maximum adsorption capacity of bovine brain galactocerebroside on samples of silicalite-1 synthesized on solid supports (in powder form and applied to glass) is determined. It changes in the range of 6 to 27 mg/g, depending on the accessibility of adsorbent pores for lipid molecules. IR spectroscopy data show that the lipid molecules are oriented with carbohydrate fragments facing the adsorbent surface. It is established that adsorption of protein molecules (BSA and hemoglobin) occurs predominantly on the lipid, and only partially on the free outer surface of the silicalite. It is concluded that adsorption alters the parameters of the porous structure.

Walking Forward with Kinesin

Active intracellular transport of organelles relies on the coordinated activities of cytoplasmic dynein and kinesin, ATP-dependent microtubule motor proteins. While axonemal dynein was discovered during the mid-1960s, it was not until the mid-1980s that kinesin was discovered by Ron Vale and colleagues, as reported in 1985. Their research demonstrated that the newly identified protein, isolated from both squid axoplasm and bovine brain, was independently capable of driving microtubule gliding or organelle movement. These findings kicked off rapid progress in the fields of physiology and neuroscience, leading to the identification of the many members of the extended kinesin superfamily, as well as detailed explorations of their biophysical properties, cellular mechanisms of action, and roles in disease.

Abstract 4111: Water content based Electrical Properties Tomography (wEPT) for modelling delivery of Tumor Treating Fields to the brain

Abstract Objective: The purpose of this study was to investigate the application of Water content based Electrical Properties Tomography for mapping electrical properties (EPs) of brain tissues in the frequency range of 100-1000 kHz. Background: TTFields are electric fields with frequencies of 100-500 kHz that disrupt mitosis. TTFields are approved for the treatment of glioblastoma multiforme. Determining the EPs of brain tissues is important for understanding how TTFields distribute within the head. The EPs of tissues are heterogeneous, especially in the region of the tumor. Therefore methods that map EPs within the brain with high spatial resolution are desired. Water content based EP tomography (wEPT) is a method that utilizes the ratio of two T1w MRI images with different relaxation times (TRs) to map EPs based on empirically derived relationships between T1, water content (WC) and EPs. wEPT has been applied to map EPs of healthy brain at 128 MHz using typical WC and EP values of healthy tissues reported in the literature to derive the empirical models. Here we adapted wEPT to map EPs in the 100-1000 kHz range utilizing in-house measurements of WC and EPs from healthy bovine and tumor-bearing rat brain tissue. Methods: The empirical model connecting MRI images, WC and EPs in the 100-1000 kHz range were created using 32 tissue samples derived from three 3 calf brains and 1 CSF sample of a pig. For each sample, T1w MRIs with TRs {700, 4000} ms were acquired and the image ratio (Ir) between the images was calculated. EPs of samples were measured using parallel plates, and WC was measured by the wet-dry weight method. Curve fitting yielded empirical models connecting Ir, WC and EP. Next, T1w MRIs of in-vivo tumor-bearing rat brains and 6 ex-vivo pieces of calf brain were acquired, and the empirical curves described above were used to map WC and EP within the rat brains and the pieces of calf brain. EPs and WC were measured on 6 small samples excised from each imaged brain. For each sample, measured values were compared to the median WC and EPs extracted from the corresponding Regions of Interest (ROIs) in the wEPT map. Results: Anatomical structures and the tumor were clearly visible in wEPT maps. WC estimated using wEPT agreed well with measurements on excised samples. There was a clear connection between EPs estimated with wEPT and the measured values. However, in some samples large differences between wEPT-derived EP values and measurements were found. In particular, the differences between tumor and healthy tissues conductivity estimated using wEPT was significantly higher than the measured difference in conductivities within the corresponding excised samples. Conclusion: wEPT maps WC in healthy and tumor brain tissues and provides information on local electrical properties at frequencies of 100-1000 kHz. Further investigation is needed to clarify the relationship between WC and EP within this frequency range. Citation Format: Catherine Tempel-Brami, Cornelia Wenger, Hadas S. Hershkovich, Moshe Giladi, Ze'ev Bomzon. Water content based Electrical Properties Tomography (wEPT) for modelling delivery of Tumor Treating Fields to the brain [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4111.

Assignment by Negative-Ion Electrospray Tandem Mass Spectrometry of the Tetrasaccharide Backbones of Monosialylated Glycans Released from Bovine Brain Gangliosides

Gangliosides, as plasma membrane-associated sialylated glycolipids, are antigenic structures and they serve as ligands for adhesion proteins of pathogens, for toxins of bacteria, and for endogenous proteins of the host. The detectability by carbohydrate-binding proteins of glycan antigens and ligands on glycolipids can be influenced by the differing lipid moieties. To investigate glycan sequences of gangliosides as recognition structures, we have underway a program of work to develop a “gangliome” microarray consisting of isolated natural gangliosides and neoglycolipids (NGLs) derived from glycans released from them, and each linked to the same lipid molecule for arraying and comparative microarray binding analyses. Here, in the first phase of our studies, we describe a strategy for high-sensitivity assignment of the tetrasaccharide backbones and application to identification of eight of monosialylated glycans released from bovine brain gangliosides. This approach is based on negative-ion electrospray mass spectrometry with collision-induced dissociation (ESI-CID-MS/MS) of the desialylated glycans. Using this strategy, we have the data on backbone regions of four minor components among the monosialo-ganglioside-derived glycans; these are of the ganglio-, lacto-, and neolacto-series.Graphical abstract

Influence of brain plasmalogen changes on gonadotropin secretion from the cultured bovine anterior pituitary cells

We recently discovered that the orphan G-protein–coupled receptor (GPR) 61 colocalized with GnRH receptors (GnRHRs) on the surface of most of bovine gonadotrophs. A recent study suggested that ethanolamine plasmalogen (PI) is a ligand for GPR61 in mouse neuroblastoma. Therefore, this study evaluated the hypothesis that PI alters LH and FSH secretion from cultured bovine anterior pituitary (AP) cells. We prepared bovine AP cells from postpubertal heifers (26 mo old) and cultured the cells for 3.5 d. We treated the cells with increasing concentrations (0, 5, 50, 500, 5,000, 50,000, or 500,000 pg/mL) of phosphoethanolamine PI (PEPI) extracted from the bovine brain, or l-α-lysophosphatidylethanolamine PI (LEPI) extracted from the bovine brain, for 5 min before either no treatment or GnRH stimulation. The medium samples were harvested 2 h after culture for LH and FSH assays. Phosphoethanolamine PI (50–500 pg/mL) stimulated (P < 0.05) the basal secretion of FSH but not LH. Phosphoethanolamine PI at 50 pg/mL also enhanced (P < 0.05) GnRH-induced FSH secretion. However, higher doses (500–500,000 pg/mL) of PEPI suppressed GnRH-induced FSH secretion. Moreover, 50 to 500,000 pg/mL PEPI suppressed GnRH-induced LH secretion. None of the tested concentrations of LEPI showed any effect on basal or GnRH-induced LH or FSH secretion. Pretreatment with Sma and Mad pathway inhibitors suppressed FSH secretion induced by PEPI, whereas an extracellular signal-regulated kinase pathway inhibitor blocked the PEPI-induced suppression of GnRH-stimulated LH secretion. Therefore, PEPI, but not LEPI, extracted from the bovine brain, alters FSH and LH secretion from cultured AP cells. Further studies are required to decide whether PEPI binds to GPR61 and whether PEPI plays an important role in the control of gonadotropin secretion from gonadotrophs.

Leonardo da Vinci: Images of the Brain

Leonardo (1452–1519) produced hundreds of anatomical drawings from at least 30 dissections he conducted over a period of several decades. He began this endeavor to more accurately portray the human body; he continued the studies to create a treatise on human anatomy. In addition, numerous animal dissections and drawings were prepared by Leonardo. In 1490–1492 Leonardo drew an image (RL 12603r, K&amp;P 32) in keeping with contemporary concepts that the cerebral ventricles and the psychic pneuma or animal spirits they contained were responsible for function of the brain. The ventricles are illustrated as three linear spaces with the optic nerves projecting to the ventricles. With this traditional view, Leonardo was perhaps the first to indicate the meningeal envelope of the optic nerves, and possibly the first to illustrate the frontal nasal sinus. Leonardo likened the coverings of the scalp and brain to layers of an onion, an analogy represented by the sectioned onion in the upper left image. Leonardo began to question connection of cranial nerves to the ventricles. This led him to prepare a wax cast of bovine cerebral ventricles to ascertain their true configuration. A hole was drilled through the underside of the skull, a syringe inserted into the 3rd ventricle and molten wax injected. For this first known example of casting of a body cavity, Leonardo applied the technique used in casting bronze statues. After the wax hardened, he removed the brain and dissected away the tissue. Leonardo then drew a series of studies on paper to illustrate the results (RL 19127r, K&amp;P 104r). A drawing of the wax cast is at bottom right of this sheet. At top left, he has superimposed the cast onto the brain. At top right, Leonardo has drawn a brain with the ventricles in place, sectioned in the mid‐line from superior to inferior. In this image, the forebrain and midbrain are fully divided. Each half is retracted laterally. Below the level of the cerebral aqueduct, the hindbrain is only partially divided and the lower hindbrain remains intact. The central figure in the lower half of the sheet is an inferior view of the intact brain. The radiating feather‐like structure centered at the pituitary is a rete mirabile—present in cattle, not humans.Even though Leonardo used a bovine specimen, the brain he has represented is human. There is a horizontal forebrain, a midbrain flexure and an inferiorly projecting hindbrain. This is indicative of an upright posture with a sub‐terminal foramen magnum as in man. A comparison of bovine and human brains in the accompanying figures indicates the proportions and configuration of brain components are human. It was not unusual at this period in anatomical history to transfer and transform structures from different species into the corresponding human structures. Human dissection was difficult to accomplish; animal dissection was not. In Rome, 1516, a disgruntled associate delivered a complaint against him to Pope Leo X and to hospital Santo Spirito, where he was performing his studies. Leonardo relocated to Amboise in the Loire valley under patronage of Francis I. He died 2 May 1519 at the chateau of Cloux. It would be several centuries before his studies were published.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The effect of the addition of cow brain powder in commercial feed on the gonadal maturity of comet goldfish (Carassius auratus auratus)

The aim of this research was to analysis the effect of addition bovine’s brain meal in artificial feed on gonad maturity and to find out the best time of gonad maturity in comet fish.This research was conducted at Fourth Building Hatchery Faculty of Fisheries and Marine Sciences Padjadjaran University on November 2014 until Januari 2015. Freeze drying of bovine brain was conducted at Research Center Inter University Bandung Institute of Technology. The research was using Completely Randomized Design (CRD) with four treatments and three replications.The treatment were 20 mg/kg, 35 mg/kg, 50mg/kg and control. The parameters of this research are Gonado Somatic Index (GSI) and egg maturity level. Addition of bovine brain meal in feed with the dose of 50 mg/kg are giving the best result until 45 days of the care time against gonad maturity of comet fish with GSI result 12.93 %, egg maturity level ripe phase 21.115 and fecundity 1520 grain/g.

The insulin receptor is expressed and functional in cultured blood-brain barrier endothelial cells but does not mediate insulin entry from blood to brain.

Insulin and its receptor are known to be present and functional in the brain. Insulin cerebrospinal fluid concentrations have been shown to correlate with plasma levels of insulin in a nonlinear fashion, indicative of a saturable transport pathway from the blood to the brain interstitial fluid. The aim of the present study was to investigate whether insulin was transported across brain endothelial cells in vitro via an insulin receptor-dependent pathway. The study showed that the insulin receptor was expressed at both the mRNA and protein levels in bovine brain endothelial cells. Luminally applied radiolabeled insulin showed insulin receptor-mediated binding to the endothelial cells. This caused a dose-dependent increase in Akt-phosphorylation, which was inhibited by coapplication of an insulin receptor inhibitor, s961, demonstrating activation of insulin receptor signaling pathways. Transport of insulin across the blood-brain barrier in vitro was low and comparable to that of a similarly sized paracellular marker. Furthermore, insulin transport was not inhibited by coapplication of an excess of unlabeled insulin or an insulin receptor inhibitor. The insulin transport and uptake studies were repeated in mouse brain endothelial cells demonstrating similar results. Although it cannot be ruled out that culture-induced changes in the cell model could have impaired a potential insulin transport mechanism, these in vitro data indicate that peripheral insulin must reach the brain parenchyma through alternative pathways rather than crossing the blood-brain barrier via receptor mediated transcytosis.

Investigation of the composition and antioxidant activity of acetone and methanol extracts of Daphne sericea L. and Daphne gnidioides L.

The compositions of methanol and acetone extracts obtained from Daphne sericea L. and Daphne gnidioides L. were investigated. The antioxidant activities of each extract were determined by various test methods. Phenolic composition profile analysis by HPLC-DAD showed that D. gnidioides extracts contain more phenolic compounds than D. sericea extracts. Among the components, syringic acid was found to be the most abundant one in D. gnidioides extracts (42.8 and 38.4mg per g dry extract of methanol and acetone, respectively). Total flavonoid, total phenolic and total carotenoid contents of methanolic D. gnidioides extracts were found to be 244.5mg rutin/g dry weight of extract, 1219.3mg GAE/g dry weight of extract and 11.9mg/g dry weight of extract, respectively. DPPH·, ABTS·+ and O2·- assays were applied to plant extracts as radical scavenging activity tests. Methanolic extracts of D. sericea and D. gnidioides showed the highest radical scavenging activities according to DPPH· and ABTS·+ tests (61.6 and 147.2 in terms of IC50, respectively). Antioxidant activity tests for measuring lipid oxidation inhibiting capacity were carried in low density lipoprotein (LDL) and bovine brain extract (BBE). Methanolic extracts of D. gnidioides and D. sericea demonstrated remarkable lipid oxidation inhibiting capacity in LDL and BBE tests.