BALB Mice Research Articles

Analysis of experimental mouse PRNP genetic polymorphisms and their susceptibility to prion diseases

Research studies showed that the polymorphisms in prion protein gene (PRNP) were associated with susceptibility to prion diseases in several animals, including humans and mouse. Several mouse strains carried natural PRNP mutations which had been identified and these could provide as animal models for human prion diseases. In this study, the genetic polymorphisms of PRNP in six common mouse strains were investigated. The experimental mice included KM mouse, ICR mouse, DBA mouse, C3H/He mouse, C57BL mouse and BALB mouse. The results showed only one new polymorphism was identified compared with the reference sequence. The identified new mutation site was C564T and it was homozygous, but this locus did not result in amino acid change. Sequence analyses suggested that these six mouse strains were susceptible to prion diseases and are suitable as susceptibility models of prion diseases.

Involvement of serotonin and oxytocin in neural mechanism regulating amicable social signal in male mice: Implication for impaired recognition of amicable cues in BALB/c strain.

Social signals play a primary role in regulating social relationships among male mice. The present series of experiments investigated the neural mechanisms underlying an induction of amicable cues that facilitate social approach in male mice of the C57BL/6 (B6) and BALB/c (BALB) strains. Male mice exhibit approach behavior and suppression of territorial scent marking toward amicable counterparts. Exposure of a group-housed mouse that maintains an amicable relationship induced social approach in B6 recipient mice, as expressed by increased preference of stay in proximity and decreased scent marks relevant to those of a single-housed mouse. Nasal administration of oxytocin (OT) to stimulus mice appeared to enhance social approach in B6 recipient mice. Systemic administration of buspirone (5-HT1A agonist) to stimulus mice also increased approach in B6 recipient mice, whereas a nasal OT antagonist infusion followed by systemic buspirone injection of stimulus mice blocked this buspirone-induced approach in B6 recipient mice. BALB mice likely possess an intact signaling system as shown in B6 mice, in which the 5-HT → OT pathway is a primary modulator for social amicable signals. However, BALB mice could not exhibit signal-dependent change in approach behavior. No impairment in olfactory discrimination or approach behavior toward social stimuli was found in BALB mice. It is concluded that social cues for facilitating social approach are eliminated via the 5-HT → OT pathway, and BALB mice as a low social strain have a deficit in recognition of specific signals associated with amicability. (PsycINFO Database Record

Extraction of biofilm produced by Escherichia coli that are isolated from animals infected with diarrhea and study its protective role

This study was conducted to detect the ability of E.coli isolated from diarrhea to produce biofilm and protection against diseases cause by these bacteria compared with whole cell sonicated antigen. One hundred two fecal samples (52 fecal samples from cows, calves and 50 fecal samples from sheep,goat) were collected from College of veterinary medicine-university of Baghdad, College of Agriculture-university of Baghdad, Dora zone and Abu-Ghraib zone. Samples were cultured on MacConkey and Eosin Methylene Blue agar and after purification of cultured bacteria, biochemical tests, API 20 E System and RapIDTM ONE System kit were done. Results showed that 91 out of 102 fecal samples have the characteristics belong to E.coli ,the ability of these isolates to produce biofilm were detected and the results showed that 38 out of 49 E.coli isolates from fecal samples of cow produce biofilm (77.55%)and 39 out of 42 E.coli isolates from fecal samples of sheep produce biofilm (92.85%) with different thickness ranged between (0.2-2)mm, while 11 isolates from 49 fecal samples of cow and calves and 39 isolates from 42 fecal samples of sheep had not produce biofilm. The results showed that protein concentration of biofilm was 92 mg/ml for one isolate and 70 mg/ ml for the whole sonicated antigen of the bacteria that produce biofilm. Three types of bacterial antigens were prepared as follow; Whole cell sonicated antigen (WCA) of biofilm producer E.coli, biofilm extract antigen (biA) with protein concentration 3.5 mg/ml and biofilm extract antigen (biA) with protein concentration 14 mg/ml, then these antigens were injected in 50 healthy White BALB mice. Results showed that survival time of animals in the immunized group with biofilm antigen of high protein concentration (14 mg/ml) was longer (652.8 hrs) than animals immunized with whole sonicated antigen of protein concentration 3.5 mg/ml antigen (378 hrs ) and than the group with biofilm antigen of protein concentration 3.5 mg/ml (513.6 hrs) , heavy bacterial isolation were recorded in the internal organs of the immunized infected animals at (12-48) hours post infection, while moderate bacterial isolation at day 30 post infection. Histopathological examination showed large abscess which caused adhesion of liver with stomach and spleen with stomach and pancreas surround by dense cellular fibrous tissue, the result showed that all animals of control positive group,5 animals of group injected with(WCA) produce biofilm,3 animals of group injected with (biA) 3.5 mg/ml and 1 animal of group injected with (biA) 14 mg/ml were died during 12-48 hours post infection, acute suppurative reaction were seen in internal organs of animals died at 12-24 hours post infection, granulomatous lesions were seen in most internal organs of animals in group 2 , 3 and 4 at day 30 post challenge, while mild inflammatory reaction was recorded in most internal organs of group 4 at day 30 post challenge.

Age-related changes in cerebellar and hypothalamic function accompany non-microglial immune gene expression, altered synapse organization, and excitatory amino acid neurotransmission deficits.

We describe age-related molecular and neuronal changes that disrupt mobility or energy balance based on brain region and genetic background. Compared to young mice, aged C57BL/6 mice exhibit marked locomotor (but not energy balance) impairments. In contrast, aged BALB mice exhibit marked energy balance (but not locomotor) impairments. Age-related changes in cerebellar or hypothalamic gene expression accompany these phenotypes. Aging evokes upregulation of immune pattern recognition receptors and cell adhesion molecules. However, these changes do not localize to microglia, the major CNS immunocyte. Consistent with a neuronal role, there is a marked age-related increase in excitatory synapses over the cerebellum and hypothalamus. Functional imaging of these regions is consistent with age-related synaptic impairments. These studies suggest that aging reactivates a developmental program employed during embryogenesis where immune molecules guide synapse formation and pruning. Renewed activity in this program may disrupt excitatory neurotransmission, causing significant behavioral deficits.

Differential innate immune cell signatures and effects regulated by toll-like receptor 4 during murine lung tumor promotion

ABSTRACTTumor promotion is an early and critical stage during lung adenocarcinoma (ADC). We previously demonstrated that Tlr4 mutant mice were more susceptible to butylated hydroxytoluene (BHT)-induced pulmonary inflammation and tumor promotion in comparison to Tlr4-sufficient mice. Our study objective was to elucidate the underlying differences in Tlr4 mutant mice in innate immune cell populations, their functional responses, and the influence of these cellular differences on ADC progenitor (type II) cells following BHT-treatment. BALB (Tlr4-sufficient) and C.C3-Tlr4Lps-d/J (BALBLpsd; Tlr4 mutant) mice were treated with BHT (promoter) followed by bronchoalveolar lavage (BAL) and flow cytometry processing on the lungs. ELISAs, Club cell enrichment, macrophage function, and RNA isolation were also performed. Bone marrow-derived macrophages (BMDM) co-cultured with a type II cell line were used for wound healing assays. Innate immune cells significantly increased in whole lung in BHT-treated BALBLpsd mice compared to BALB mice. BHT-treated BALBLpsd mice demonstrated enhanced macrophage functionality, increased epithelial wound closure via BMDMs, and increased Club cell number in BALBLpsd mice, all compared to BALB BHT-treated mice. Cytokine/chemokine (Kc, Mcp1) and growth factor (Igf1) levels also significantly differed among the strains and within macrophages, gene expression, and cell surface markers collectively demonstrated a more plastic phenotype in BALBLpsd mice. Therefore, these correlative studies suggest that distinct innate immune cell populations are associated with the differences observed in the Tlr4-mutant model. Future studies will investigate the macrophage origins and the utility of the pathways identified herein as indicators of immune system deficiencies and lung tumorigenesis.

Platelet-derived Growth Factor-B Protects Rat Cardiac Allografts From Ischemia-reperfusion Injury.

Microvascular dysfunction and cardiomyocyte injury are hallmarks of ischemia-reperfusion injury (IRI) after heart transplantation. Platelet-derived growth factors (PDGF) have an ambiguous role in this deleterious cascade. On one hand, PDGF may exert vascular stabilizing and antiapoptotic actions through endothelial-pericyte and endothelial-cardiomyocyte crosstalk in the heart; and on the other hand, PDGF signaling mediates neointimal formation and exacerbates chronic rejection in cardiac allografts. The balance between these potentially harmful and beneficial actions determines the final outcome of cardiac allografts. We transplanted cardiac allografts from Dark Agouti rat and Balb mouse donors to fully major histocompatibility complex-mismatched Wistar Furth rat or C57 mouse recipients with a clinically relevant 2-hour cold ischemia and 1-hour warm ischemia. Ex vivo intracoronary delivery of adenovirus-mediated gene transfer of recombinant human PDGF-BB upregulated messenger RNA expression of anti-mesenchymal transition and survival factors BMP-7 and Bcl-2 and preserved capillary density in rat cardiac allografts at day 10. In mouse cardiac allografts PDGF receptor-β, but not -α intragraft messenger RNA levels were reduced and capillary protein localization was lost during IRI. The PDGF receptor tyrosine kinase inhibitor imatinib mesylate and a monoclonal antibody against PDGF receptor-α enhanced myocardial damage evidenced by serum cardiac troponin T release in the rat and mouse cardiac allografts 6 hours after reperfusion, respectively. Moreover, imatinib mesylate enhanced rat cardiac allograft vasculopathy, cardiac fibrosis, and late allograft loss at day 56. Our results suggest that PDGF-B signaling may play a role in endothelial and cardiomyocyte recovery from IRI after heart transplantation.

The Th17 pathway in the peripheral lung microenvironment interacts with expression of collagen V in the late state of experimental pulmonary fibrosis

BackgroundMyofibroblasts derived from fibroblasts in the pathogenesis of pulmonary fibrosis causes excessive and disordered deposition of matrix proteins, including collagen V, which can cause a Th17-mediated immune response and lead to apoptosis. However, whether the intrinsic ability of lung FBs to produce the matrix depends on their site-specific variations is not known. AimTo investigate the link between Th17 and collagen V that maintains pulmonary remodeling in the peripheral lung microenvironment during the late stage of experimental pulmonary fibrosis. MethodsYoung male mice including wild Balb/c mice (BALB, n=10), wild C57 Black/6J mice (C57, n=10) and IL-17 receptor A knockout mice (KO, n=8), were sacrificed 21 days after treatment with bleomycin. Picrosirius red staining, immunohistochemistry for IL-17-related markers and “in situ” detection of apoptosis, immunofluorescence for collagen types I and V, primary cell cultures from tissue lung explants for RT-PCR and electron microscopy were used. ResultsThe peripheral deposition of extracellular matrix components by myofibroblasts during the late stage is maintained in C57 mice compared with that in Balb mice and is not changed in the absence of IL-17 receptor A; however, the absence of IL-17 receptor A induces overexpression of type V collagen, amplifies the peripheral expression of IL-17 and IL-17-related cytokines and reduces peripheral lung fibroblast apoptosis. ConclusionA positive feedback loop between the expression patterns of collagen V and IL-17 may coordinate the maintenance of peripheral collagen I in the absence of IL-17 receptor A in fibrosis-susceptible strains in a site-specific manner.

Ehrlich tumor inhibition using doxorubicin containing liposomes

Ehrlich tumors were grown in female balb mice by subcutaneous injection of Ehrlich ascites carcinoma cells. Mice bearing Ehrlich tumor were injected with saline, DOX in solution or DOX encapsulated within liposomes prepared from DMPC/CHOL/DPPG/PEG-PE (100:100:60:4) in molar ratio. Cytotoxicity assay showed that the IC50 of liposomes containing DOX was greater than that DOX only. Tumor growth inhibition curves in terms of mean tumor size (cm3) were presented. All the DOX formulations were effective in preventing tumor growth compared to saline. Treatment with DOX loaded liposomes displayed a pronounced inhibition in tumor growth than treatment with DOX only. Histopathological examination of the entire tumor sections for the various groups revealed marked differences in cellular features accompanied by varying degrees in necrosis percentage ranging from 12% for saline treated mice to 70% for DOX loaded liposome treated mice. The proposed liposomal formulation can efficiently deliver the drug into the tumor cells by endocytosis (or passive diffusion) and lead to a high concentration of DOX in the tumor cells. The study showed that the formulation of liposomal doxorubicin improved the therapeutic index of DOX and had increased anti-tumor activity against Ehrlich tumor models.

Mesoporous silica coated gold nanorods loaded doxorubicin for combined chemo-photothermal therapy.

The efficacy of the combined chemo-photothermal therapy, using a mesoporous silica-coated gold nanorods loaded DOX (pGNRs@mSiO2-DOX), was consistently tested both in vitro and in vivo. The prepared nanoparticles that were characterized using transmission electron microscopy (TEM), UV-vis absorption spectroscopy and zeta potential showed high doxorubicin loading capacity in addition to its pH-responsive release. The pGNRs@mSiO2-DOX photo-heat conversion characteristic found to be stable for several repeated NIR irradiated doses was tested in simulated body fluid. In vitro results showed that pGNRs@mSiO2-DOX causes a significant damage in breast cancer cell line MCF-7 compared to free DOX. Contrary to this, it showed low toxicity to human amnion wish cells compared to CTAB coated GNRs and free DOX. In vivo results showed that intravenous administration of pGNRs@mSiO2-DOX (1.7 mg/kg) markedly suppresses the growth of subcutaneous Ehrlich carcinoma in female Balb mice (p<0.0001). Consistently, histopathological examination revealed a complete loss of tumor cellular details for mice that received the combined treatment. Based on the obtained results, this passively targeted pGNRs@mSiO2-DOX could specifically deliver drug and excessive local heat to tumor sites achieving high combined therapeutic efficacy.

Escitalopram or Novel Herbal Mixture Treatments during or following Exposure to Stress Reduce Anxiety-Like Behavior through Corticosterone and BDNF Modifications

Anxiety disorders are a major public health concern worldwide. Studies indicate that repeated exposure to adverse experiences early in life can lead to anxiety disorders in adulthood. Current treatments for anxiety disorders are characterized by a low success rate and are associated with a wide variety of side effects. The aim of the present study was to evaluate the anxiolytic effects of a novel herbal treatment, in comparison to treatment with the selective serotonin reuptake inhibitor escitalopram. We recently demonstrated the anxiolytic effects of these treatments in BALB mice previously exposed to one week of stress. In the present study, ICR mice were exposed to post natal maternal separation and to 4 weeks of unpredictable chronic mild stress in adolescence, and treated during or following exposure to stress with the novel herbal treatment or with escitalopram. Anxiety-like behavior was evaluated in the elevated plus maze. Blood corticosterone levels were evaluated using radioimmunoassay. Brain derived neurotrophic factor levels in the hippocampus were evaluated using enzyme-linked immunosorbent assay. We found that (1) exposure to stress in childhood and adolescence increased anxiety-like behavior in adulthood; (2) the herbal treatment reduced anxiety-like behavior, both when treated during or following exposure to stress; (3) blood corticosterone levels were reduced following treatment with the herbal treatment or escitalopram, when treated during or following exposure to stress; (4) brain derived neurotrophic factor levels in the hippocampus of mice treated with the herbal treatment or escitalopram were increased, when treated either during or following exposure to stress. This study expands our previous findings and further points to the proposed herbal compound's potential to be highly efficacious in treating anxiety disorders in humans.

Dopamine D1 and opioid receptor antagonism effects on the acquisition and expression of fat-conditioned flavor preferences in BALB/c and SWR mice

Sugar and fat appetites are influenced by unlearned and learned responses to orosensory and post-ingestive properties which are mediated by dopamine (DA) and opioid transmitter systems. In BALB and SWR mice, acquisition and expression of sucrose conditioned flavor preferences (CFP) are differentially affected by systemic DA D1 (SCH23390: SCH) and opioid (naltrexone: NTX) antagonism. The present study examined whether fat-CFP occurred in these strains using preferred (5%) and less preferred (0.5%) Intralipid solutions, and how SCH and NTX altered its expression and acquisition. Food-restricted mice drank flavored (CS+, e.g., cherry) 5% and flavored (CS−, e.g., grape) 0.5% Intralipid solutions in alternate 1h/day sessions. Six two-bottle tests with both CS flavors presented in 0.5% Intralipid occurred. Robust and comparable fat-CFP was observed in both strains. In Experiment 2, vehicle (Veh), SCH (50–800nmol/kg) or NTX (1–5mg/kg) were administered prior to two-bottle testing in an identical paradigm. Expression of fat-CFP was significantly reduced by SCH and NTX in BALB mice, and only by SCH in SWR mice. NTX produced significantly greater inhibition of fat-CFP expression in BALB versus SWR mice. In Experiment 3, separate groups of BALB and SWR mice received Veh, SCH (50nmol/kg) or NTX (1mg/kg) injections prior to daily one-bottle CS+ and CS− training sessions to assess acquisition effects. Six two-bottle CS+ vs. CS− choice tests were then conducted. Fat-CFP acquisition was significantly reduced in SWR mice by SCH, but not NTX, and was only mildly affected by both antagonists in BALB mice. SCH produced significantly greater inhibition of fat-CFP acquisition in SWR versus BALB mice. The pattern of SCH effects upon the expression and acquisition of fat-CFP in BALB and SWR mice was quite similar to that observed for sucrose-CFP, suggesting similar DA D1 receptor involvement in sugar- and fat-CFP in these mouse strains.

Abstract 3607: Toll-like receptor (TLR) 4 and innate immune cells are involved in the early stages of murine lung tumor promotion.

Abstract Chronic inflammatory lung diseases predispose to lung neoplasia. Activation of certain innate immune pathways may confer protection against lung adenocarcinoma (AC) development. Lack of the innate immunity toll-like receptor 4 (Tlr4) gene enhances susceptibility to lung inflammation via bacterial infections while its presence can mediate anti-cancer efficacy. We previously found that Tlr4 is protective in butlylated hydroxytoulene (BHT)-induced pulmonary chronic inflammation and turmorigenesis and using transcriptomics, analyzed the downstream mechanisms involved (Bauer et al., 2005, 2009). The objective of this study was to further characterize the early stage of tumor promotion by determining the phenotypes of the immune cells in the lungs and the interactions between the alveolar immune cells and pulmonary epithelial cells. C.C3H-Tlr4Lps-d(BALBLps-d;Tlr4 mutant) and BALB/c (BALB;Tlr4 wild-type) mice were chronically injected weekly for a min. of 4 weeks with BHT or oil vehicle. Mice were then euthanized at several time points (1, 3, and 30 dy) following the last injection. Subsequently, lungs were half-lung bronchoalveolar lavaged (BAL) for BAL fluid (BALF), while the unlavaged lung was snap frozen. BALF was analyzed for cellular infiltrates by differentials and protein content (indicative of hyperpermeability). Staining and flow cytometric analysis was performed on the same lungs using markers to differentiate between lymphocytes, neutrophils and several pulmonary macrophage (MΦ) populations, including alveolar, interstitial, and activated macrophages. BHT-induced chronic pulmonary inflammation in the BALBLps-d mice had significantly elevated levels of inflammatory cells and hyperpermeability during the early stages of promotion above that observed in the BALB mice. Results from the flow cytometric analysis suggest that there are increased numbers of MΦs and neutrophils in the lungs of BALBLps-d mice compared to the BALB mice. In further phenotyping of the MΦs via flow cytometry, alternatively activated MΦs (M2; CD206+), were found in greater abundance than classically activated MΦs (M1, iNOS+) in both strains with an increase in BALBLps-d vs. BALB mice. These findings, along with several genes upregulated in the BALBLps-d but not the BALB mice (Pparg, Arg1, Ccl17), suggest that the absence of TLR4 function may lead to an increase in the M2 phenotype, which has been found to assist tumorgenesis. The results demonstrate the importance of TLR4 in the immune response against BHT-induced lung inflammation and subsequently lung tumorgenesis. Future experiments will further investigate the interactions between pulmonary MΦs and pulmonary epithelial cells. Funded by ACS RSG-10-162-01-LIB (AKB). Citation Format: Carla-Maria Alexander, Thomas Hill, Ross Osgood, Alison K. Bauer. Toll-like receptor (TLR) 4 and innate immune cells are involved in the early stages of murine lung tumor promotion. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3607. doi:10.1158/1538-7445.AM2013-3607

Abstract 4801: Bronchoalveolar lavage fluid (BALF) from subchronic exposure to butylated-hydroxytoluene (BHT) inhibits gap junctional communication in a toll-like receptor 4-dependent manner.

Abstract Subchronic dosing of BHT in mice is a well-known model used to discretely study the tumor promotion stage of lung adenocarcinoma. Using this model, we have previously shown that functional toll-like receptor 4 (TLR4) is protective against inflammatory sequelae and primary tumor formation in the mouse lung. Gap junctional intercellular communication (GJIC) has been shown to regulate the signal transduction pathways of cells, and inhibition of this communication by toxicant exposure has been linked to dysregulation and eventual carcinogenic endpoints. As part of our research on the protective nature of TLR4 during tumor promotion, we utilized the BHT subchronic model (4 weekly i.p. doses of BHT) to examine the early effects on BALF in both BALB/c (BALB; Tlr4 sufficient) and C.C3H-Tlr4Lps-d mice (BALBLps-d-d; Tlr4 mutant). Analysis of the BALF 24 hours after the final dose for protein content and cytology was indicative of an inflammatory profile in all mice treated with BHT. Magnitude of response to BHT was highest in the BALBLps-d mice; the BALB appeared resistant. In an effort to further characterize the response of the pulmonary epithelium, the BALF samples were used to treat cultured C10 cells, a murine immortalized line derived from BALB/c alveolar type II cells, which are a precursor cell type for adenocarcinoma. Using the scrape-load dye transfer assay, a well-defined method to measure GJIC, we demonstrated marked inhibition of cellular communication after a 4 hour exposure to cell-free BALF that is both treatment and strain dependent. The direction and magnitude of effect is in parallel with that of the direct analysis of the BALF, i.e. we observed more inhibition in BALF from BALBLps-d compared to BALB mice. In addition, quantitative PCR (QPCR) analysis of C10 cells exposed to cell-free BALF for 24 hours indicates alterations in the transcription patterns in genes of innate immunity that are also treatment and strain dependent. These novel findings suggest endocrine signaling from the inflammatory milieu are complicit in the inhibition of GJIC during the promotion phase of carcinogenesis. Furthermore, these data establish a link between active gap junctions, tumor promotion, and the protective nature of TLR4 in an ex vivo model which will facilitate a mechanistic examination of these protective effects at the molecular level. Funded by ACS RSG-10-162-01-LIB (AKB). Citation Format: Thomas Hill, Carla-Maria Alexander, Alison K. Bauer. Bronchoalveolar lavage fluid (BALF) from subchronic exposure to butylated-hydroxytoluene (BHT) inhibits gap junctional communication in a toll-like receptor 4-dependent manner. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4801. doi:10.1158/1538-7445.AM2013-4801

Increased Resistance to Staphylococcus aureus Endophthalmitis in BALB/c Mice: Fas Ligand Is Required for Resolution of Inflammation but Not for Bacterial Clearance

FasL was recently shown be required for bacterial clearance in C57BL/6 mice that express the FasL.1 allotype. The FasL.2 allotype is expressed in BALB/c mice and exhibits increased binding affinity to and increased cytotoxic activity against Fas(+) target cells. Therefore, we hypothesized that BALB/c mice would be more resistant to Staphylococcus aureus-induced endophthalmitis. To test this hypothesis, C57BL/6, BALB/c, and BALB(gld) mice received intravitreal injections of 2,500 CFU of S. aureus (RN6390). Clinical examinations, electroretinography (ERG), histology, and bacterial quantification were performed at 24, 48, 72, and 96 h postinjection. The myeloperoxidase (MPO) assay was used to quantitate neutrophil infiltration. At 96 h postinfection, 86% of C57BL/6 mice presented with complete destruction of the eye, compared to only 29% of BALB/c mice with complete destruction. To our surprise, in the absence of Fas ligand, BALB(gld) mice showed no difference in bacterial clearance compared to BALB/c mice. However, histology and ERG analysis revealed increased retinal damage and significant loss of retinal function. MPO analysis revealed equal numbers of neutrophils in BALB(gld) and BALB/c mice at 24 h postinfection. However, at 48 h, the neutrophil numbers remained significantly elevated in BALB(gld) mice, correlating with the increased retinal damage observed in BALB(gld) mice. We conclude that the increased resistance to S. aureus induced endophthalmitis in BALB/c mice is not dependent upon the FasL. However, in contrast to C57BL/6 mice, FasL is required for resolution of inflammation and protecting host tissue from nonspecific damage in BALB/c mice.

Preparation and Characterization of SiO&lt;SUB&gt;2&lt;/SUB&gt;–Au Nanoshells: &lt;I&gt;In&lt;/I&gt; &lt;I&gt;Vivo&lt;/I&gt; Study of Its Photo-Heat Conversion

In the event of cancer treatment, photothermal therapy has met successful cancerous cells damage with highly reduced toxicity to normal cells. The prepared GNSs samples have been characterized using transmission electron microscope (TEM), dynamic light scattering, zeta potential and UV-VIS absorption spectroscopy. In-vivo photo-heat conversion of GNSs accumulated in Ehrlich tumor cells inoculated in female balb mice was monitored by measuring tumor tissue temperature as a function of NIR laser exposure time. Resultant heating and therapeutic efficacy were assessed by monitoring tumor growth/regression and tumor cells necrotic percentage. Histopathological examinations for treated and control tumors using light microscope and transmission electron microscopes (TEM) were performed to evaluate the treatment effects. Passively targeted pegylated gold nanoshells were found to have localized photo-heat conversion sufficient to selectively destruct tumor cells. This has been emphasized by the significant decrease in Ehrlich tumor volume for treated groups that administrated either intratumorly (IT) or intravenously (IV) with GNSs. Light microscope examinations revealed high necrotic percentages for both administration routes. TEM images showed degenerated cell membrane and nuclear envelop as well as the appearance of nucleus debris and other cell organelles. This non-invasive protocol showed great promise as a technique for selective cancer photo-thermal therapy.

Bronchoalveolar Lavage Fluid Utilized Ex Vivo to Validate In Vivo Findings: Inhibition of Gap Junction Activity in Lung Tumor Promotion is Toll-Like Receptor 4-Dependent.

TLR4 protects against lung tumor promotion and pulmonary inflammation in mice. Connexin 43 (Cx43), a gap junction gene, was increased in Tlr4 wildtype compared to Tlr4-mutant mice in response to promotion, which suggests gap junctional intercellular communication (GJIC) may be compromised. We hypothesized that the early tumor microenvironment, represented by Bronchoalveolar Lavage Fluid (BALF) from Butylated hydroxytoluene (BHT; promoter)-treated mice, would produce TLR4-dependent changes in pulmonary epithelium, including dysregulation of GJIC in the Tlr4-mutant (BALBLps-d) compared to the Tlr4-sufficient (BALB; wildtype) mice. BHT (4 weekly doses) was injected ip followed by BALF collection at 24 h. BALF total protein and total macrophages were significantly elevated in BHT-treated BALBLps-d over BALB mice, similar to previous findings. BALF was then utilized in an ex vivo manner to treat C10 cells, a murine alveolar type II cell line, followed by the scrape-load dye transfer assay (GJIC), Cx43 immunostaining, and quantitative RT-PCR (Mcp-1, monocyte chemotactic protein 1). GJIC was markedly reduced in C10 cells treated with BHT-treated BALBLps-d BALF for 4 and 24 h compared to BALB and control BALF from the respective mice (p < 0.05). Mcp-1, a chemokine, was also significantly increased in the BHT-treated BALBLps-d BALF compared to the BALB mice, and Cx43 protein expression in the cell membrane altered. These novel findings suggest signaling from the BALF milieu is involved in GJIC dysregulation associated with promotion and links gap junctions to pulmonary TLR4 protection in a novel ex vivo model that could assist in future potential tumor promoter screening.

Potential role of Atp5g3 in epigenetic regulation of alcohol preference or obesity from a mouse genomic perspective

The mitochondrial ATP synthase, subunit c, isoform 3 gene (Atp5g3) encodes subunit 9, the subunit of the multisubunit enzyme that catalyzes ATP synthesis during oxidative phosphorylation in mitochondria. According to the Ensembl database, Atp5g3 in mice is located on chromosome 2 between 73746504 and 73749383 bp, within the genomic regions of two sets of quantitative trait loci - alcohol preference and body weight. Both of those traits are more influenced by epigenetic factors than many other traits are. Using currently available phenotype and gene expression profiles from the GeneNetwork database, we obtained correlations between Atp5g3 and alcoholism- and obesity-relevant phenotypes. The correlation in expression levels between Atp5g3 and each of its 12 partner genes in the molecular interaction are different in various tissues and genes. Transcriptome mapping indicated that Atp5g3 is differentially regulated in the hippocampus, cerebellum, and liver. Owing to a lack of known polymorphisms of Atp5g3 among three relevant mouse strains, C57BL/6J (B6), DBA/2J (D2), and BALB/ cJ, the molecular mechanism for the connection between Atp5g3 and alcoholism and body weight requires further investigation.

Biodistribution of ultra small superparamagnetic iron oxide nanoparticles in BALB mice

Recently ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles (NPs) have been widely used for medical applications. One of their important applications is using these particles as MRI contrast agent. While various research works have been done about MRI application of USPIOs, there is limited research about their uptakes in various organs. The aim of this study was to evaluate the biodistribution of dextran coated iron oxide NPs labelled with 99mTc in various organs via intravenous injection in Balb/c mice. The magnetite NPs were dispersed in phosphate buffered saline and SnCl2 which was used as a reduction reagent. Subsequently, the radioisotope 99mTc was mixed directly into the reaction solution. The labeling efficiency of USPIOs labeled with 99mTc, was above 99 %. Sixty mice were sacrificed at 12 different time points (From 1 min to 48 h post injections; five mice at each time). The percentage of injected dose per gram of each organ was measured by direct counting for 19 harvested organs of the mice. The biodistribution of 99mTc-USPIO in Balb/c mice showed dramatic uptake in reticuloendothelial system. Accordingly, about 75 percent of injected dose was found in spleen and liver at 15 min post injection. More than 24 % of the NPs remain in liver after 48 h post-injection and their clearance is so fast in other organs. The results suggest that USPIOs as characterized in our study can be potentially used as contrast agent in MR Imaging, distributing reticuloendothelial system specially spleen and liver.

N,N,N-Trimethyl chitosan nanoparticles for controlled intranasal delivery of HBV surface antigen

Hepatitis B virus surface antigen (HBsAg) loaded N,N,N-trimethyl chitosan nanoparticles (N-TMC NPs) were formulated and studied for controlled intranasal delivery. The size and surface properties of the NPs can be tuned by modifying the concentration of N-TMC and found to be 66±13, 76±9nm for 0.25 and 0.5wt.% respectively. Loading of 380 and 760μl of HBsAg yielded 143±33, 259±47nm sized spherical N-TMC NPs with highest loading efficiency and capacity of 90–93%, and 96–97% respectively. In vitro drug release analysis ensured 93% cumulative release of HBsAg antigen over prolonged period (43days). In vivo immunological study was performed using 6–8weeks old female BALB mice and reveals adjuvants efficiency of NPs for antigen is highly stable and better than standard. Obtained results show that N-TMC NPs can be extensively used in controlled intra nasal delivery to treat various diseases including hepatitis B and allergic rhinitis.

Comparison of focally induced epileptiform activities in C57BL/6 and BALB/c mice by using in vivo EEG recording

Epilepsy characterized by repeated seizures is influenced by genetic factors. Seizure response of inbred mouse strains changes depending on the variety of stimuli including chemical (e.g., pentylenetetrazole, nicotine, cocaine, NMDA, kainate), physical (e.g., auditory) or electrical. In this study, we compared the susceptibilities of C57BL/6 and BALB/c mice strains to penicillin induced epileptiform activity (a focally induced, experimental epilepsy model), by analyzing the spike onset latency, spike amplitude and spike frequency. The power spectrums of baseline EEGs were also investigated. We found no alterations of spike onset latencies between the C57 and BALB mice. However, spike amplitudes and spike frequencies were found to be higher in BALB mice than C57 mice. With regard to EEG power spectrum, absolute power of investigated bands was not different between the two strains. Interestingly, the relative power of all investigated bands differed significantly between two strains. The relative power of delta and theta was lower whereas relative power of alpha, beta and gamma was higher in C57 mice compared to BALB mice. In conclusion, our findings showed that BALB mice are more sensitive to penicillin induced epileptiform activity when compared to C57 strain.