Female BALBc Mice Research Articles

Efficacy of a saline wash plus vancomycin/tobramycin-doped PVA composite (PVA-VAN/TOB-P) in a mouse pouch infection model implanted with 3D-printed porous titanium cylinders.

The efficacy of saline irrigation for treatment of implant-associated infections is limited in the presence of porous metallic implants. This study evaluated the therapeutic efficacy of antibiotic doped bioceramic (vancomycin/tobramycin-doped polyvinyl alcohol composite (PVA-VAN/TOB-P)) after saline wash in a mouse infection model implanted with titanium cylinders. Air pouches created in female BalBc mice by subcutaneous injection of air. In the first of two independent studies, pouches were implanted with titanium cylinders (400, 700, and 100 µm pore sizes) and inoculated with Staphylococcus aureus (1 × 103 or 1 × 106 colony-forming units (CFU)/pouch) to establish infection and biofilm formation. Mice were killed after one week for microbiological analysis. In the second study, pouches were implanted with 400 µm titanium cylinders and inoculated with S. aureus (1 × 103 or 1 × 106 CFU/pouch). Four groups were tested: 1) no bacteria; 2) bacteria without saline wash; 3) saline wash only; and 4) saline wash plus PVA-VAN/TOB-P. After seven days, the pouches were opened and washed with saline alone, or had an additional injection of PVA-VAN/TOB-P. Mice were killed 14 days after pouch wash. The first part of the study showed that low-grade infection was more significant in 400 µm cylinders than cylinders with larger pore sizes (p < 0.05). The second part of the study showed that saline wash alone was ineffective in eradicating both low- and high-grade infections. Saline plus PVA-VAN/TOB-P eradicated the titanium cylinder-associated infections, as manifested by negative cultures of the washouts and supported by scanning electron microscopy and histology. Porous titanium cylinders were vulnerable to bacterial infection and biofilm formation that could not be treated by saline irrigation alone. Application of PVA-VAN/TOB-P directly into the surgical site alone or after saline wash represents a feasible approach for prevention and/or treatment of porous implant-related infections.

Effects of Boric acid as Maternal Feed Additives on the Development and Sex Ratio of Mouse pups

Boron is primarily used in industrial applications, with recent interest revolving around its effects on metabolism. In this study, we administered boric acid (BA), which has positive effects on reproduction, in conjunction with feed supplementation to serve as a model for experimental animal development and breeding. The pregnancy performance, offspring development, and biochemical effects of mice given feed supplemented with BA at concentrations of 0 (control group), 250, and 500 ppm (BA groups) were investigated. A total of 18 female Balb-C mice were utilized for pregnancy. The mice were given the BA-supplemented feed during a period encompassing three weeks of pregnancy and three weeks of lactation. The numbers and weights of offspring born in cages on days 19–21 were determined. Blood and tissue samples were collected from the offspring during the third week postnatal, and the malondialdehyde (MDA) and total antioxidant and oxidant status (TAS, TOS, and OSI) levels were determined. A significant increase in female offspring was observed in the groups born to mice fed with BA compared to the control group. Positive development in organ weights was observed in the 250-ppm BA group. The 250-ppm group exhibited a significant increase in TAS compared to the control group, while TOS and MDA levels showed a decrease. Also, the levels of BA groups were found to decrease in both the OSI index serum and organ samples compared to the control group. Thus, the use of 250-ppm BA demonstrated positive effects on female offspring production, organ development, and antioxidant levels.

The effects of MEK and PKA inhibition on Spire proteins during oocyte maturation.

Asymmetric division of oocytes driven by chromosome migration is a crucial step of oocyte maturation. Actin filaments take key roles in chromosome migration in oocytesThe aim of this study was to determine the effects of MEK and PKA inhibition on the levels of Spire-1 and Spire-2 proteins that are known to be related to actin nucleation.MEK inhibitor PD98059 and PKA inhibitor H89 were applied during IVM to the oocytes retrieved from preovulatory ovarian follicles of PMSG induced 3-5 weeks old female BalbC mice. GVBD and PBE rates were determined. Spire-1 and Spire-2 proteins were detected by immunofluorescence and western blot in oocytes at different maturation stages.Though GVBD rates were similar in different groups, PBE rates were lower in the MEK inhibition group. Through immunofluorescence, cortical localizations of Spire-1 and Spire-2 were determined. MEK inhibition resulted in a decrease in cortical Spire-1 and Spire-2 levels in PBE oocytes. PKA inhibition led to an increase in cortical Spire-1 levels in spindle migration stage oocytes, and an increase in cortical and total Spire-2 levels in PBE oocytes. Application of both MEK and PKA inhibition resulted in compensation of the decrease in Spire-1, while Spire-2 levels remained low with no compensation of PKA inhibition.According to the results of this study, chemical inhibition of MEK and PKA during oocyte maturation alters Spire-1 and Spire-2 protein levels.

Coinfection with Schistosoma mansoni Enhances Disease Severity in Human African Trypanosomiasis.

Human African trypanosomiasis (HAT) and schistosomiasis are neglected parasitic diseases found in the African continent. This study was conducted to determine how primary infection with Schistosoma mansoni affects HAT disease progression with a secondary infection with Trypanosoma brucei rhodesiense (T.b.r) in a mouse model. Female BALB-c mice (6-8 weeks old) were randomly divided into four groups of 12 mice each. The different groups were infected with Schistosoma mansoni (100 cercariae) and Trypanosoma brucei rhodesiense (5.0 × 104) separately or together. Twenty-one days after infection with T.b.r, mice were sacrificed and samples were collected for analysis. The primary infection with S. mansoni significantly enhanced successive infection by the T.b.r; consequently, promoting HAT disease severity and curtailing host survival time. T.b.r-induced impairment of the neurological integrity and breach of the blood-brain barrier were markedly pronounced on coinfection with S. mansoni. Coinfection with S. mansoni and T.b.r resulted in microcytic hypochromic anemia characterized by the suppression of RBCs, hematocrit, hemoglobin, and red cell indices. Moreover, coinfection of the mice with the two parasites resulted in leukocytosis which was accompanied by the elevation of basophils, neutrophils, lymphocytes, monocytes, and eosinophils. More importantly, coinfection resulted in a significant elevation of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin, creatinine, urea, and uric acid, which are the markers of liver and kidney damage. Meanwhile, S. mansoni-driven dyslipidemia was significantly enhanced by the coinfection of mice with T.b.r. Moreover, coinfection with S. mansoni and T.b.r led to a strong immune response characterized by a significant increase in serum TNF-α and IFN-γ. T.b.r infection enhanced S. mansoni-induced depletion of cellular-reduced glutathione (GSH) in the brain and liver tissues, indicative of lethal oxidative damage. Similarly, coinfection resulted in a significant rise in nitric oxide (NO) and malondialdehyde (MDA) levels. Primary infection with S. mansoni exacerbates disease severity of secondary infection with T.b.r in a mouse model that is associated with harmful inflammatory response, oxidative stress, and organ injury.

The Effect of 4 Weeks Aerobic Exercise Training with Detraining Courses in Various Prevention Phases on BCl-2 and BAX Genes Expression and Proteins.

The aim of this study was to investigate the effect of aerobic exercise with detraining in different phases of prevention on BCL2 Associated X (BAX) and B-cell lymphoma 2 (BCl-2) gene expression and proteins. For this purpose, 32 female Balb-c mice (18-20 g) were purchased and randomly assigned to primordial prevention (A), primary prevention (B), secondary prevention (C), and control (D). A group performed aerobic exercise for 4 weeks, after 4T1 cells injection detrained for 8 weeks. Group B performed aerobic exercise for 4 weeks immediately after injecting 4T1 cells and then detrained for 4 weeks. In C group, the 4T1 cells were first injected and did not perform any activity for 4 weeks, followed by 4 weeks of aerobic exercise. Forty-eight hours after the last training session and detraining courses, after anesthesia, sacrificing, and tissue removal, were performed. Reverse transcription polymerase chain reaction (RT PCR) was used to measure gene expression and Western blot (WB) was used to measure protein content. One-way Analysis of variance (ANOVA) test was used to analyze data. The results showed that aerobic exercise in A, B, and C groups compared to D group reduced BCl-2 gene expression and protein and increased BAX gene expression and protein. Therefore, exercise can cause apoptosis in tumor cells by increasing pre-apoptotic factors and decreasing antiapoptotic factors in tumor cells, and consequently improving the disease status.

Therapeutic Trial of Modified Levofloxacin (MOLVX) Against Induced Colorectal Aberrant Crypt Foci (Precancerous Tissue) in Mice

&#x0D; &#x0D; &#x0D; &#x0D; Colorectal cancer ranks as the third most commonly diagnosed cancer and the second leading cause of cancer-related deaths globally. ‎Fluoroquinolones, particularly Levofloxacin, have garnered interest for their potential anti-cancer effects, primarily due to their high affinity ‎for metallic ions like copper. This affinity enhances their spectrum of activity and enables greater interaction with DNA in cancer cells, ‎thereby inhibiting proliferation. Building on our prior work, where we synthesized copper-modified Levofloxacin (MOLVX), this study ‎explores its therapeutic potential in treating precancerous colorectal lesions, known as Aberrant Crypt Foci (ACF), in a murine model. Sixty ‎female Balb-C mice were randomized into six groups (n=10 per group). The first group served as a negative control and received no treatment. ‎The remaining groups were administered azoxymethane (AOM) at 10 mg/kg body weight (BW) twice weekly to induce ACF. Among these, the ‎second group acted as a positive control and received no further treatment. The third group was intraperitoneally administered doxorubicin at ‎‎16.2 mg/kg BW once a week for four weeks. The fourth group was treated with Levofloxacin at 25 mg/kg BW via oral lavage daily for four ‎weeks. The final two groups received daily oral lavage treatments of MOLVX at doses of 5 mg/kg and 2.5 mg/kg BW for four weeks, ‎respectively. Histopathological analysis of ACF tissues, stained with hematoxylin and eosin, revealed a statistically significant reduction ‎‎(P&lt;0.05) in ACF count among groups treated with MOLVX, doxorubicin, and Levofloxacin, compared to the positive control. Additionally, both ‎MOLVX-treated groups showed a significant decrease (P&lt;0.05) in proliferating cellular antigen (PCNA) levels. In conclusion, both MOLVX ‎dosages (2.5 mg/kg and 5 mg/kg BW) demonstrated promising therapeutic efficacy against precancerous colorectal lesions in mice. However, ‎the 5 mg/kg BW dose appeared to be more potent.&#x0D; &#x0D; &#x0D; &#x0D;

P02-07 Integrated approach to evaluate (immuno)toxicity of BDE-47 in female Balb-c mice

P02-07 Integrated approach to evaluate (immuno)toxicity of BDE-47 in female Balb-c mice

Kontrollü Ovaryen Hiperstimulasyon Uterusta Apelin ve Apelin Reseptör Ekspresyonunu Arttırır

Objective; In this study, we investigated whether superovulation, which is frequently used in the clinic as controlled ovarian hyperstimulation, affect the expression of Apelin (APLN) and Apelin Receptor (APJ) in the uterus.&#x0D; &#x0D; Methods: Twelve female Balb-C mice 6-8 weeks old were used in this study. The groups were established as follows; virgin control female group (Knt) which did not receive any treatment, the pregnant control group (GK) which was on their pregnancy day 1, and the PMSG+hCG group (P+H) that received 5 IU/mouse/intraperitoneal (ip) PMSG and 5 IU/mouse/ip hCG . Vaginal plaque control was performed on the day after mating in the P+H and GK groups, and vaginal plaque-positive mice were accepted at day 0.5 of pregnancy. Mice in the P+H group were sacrificed 42 hours after hCG injection. Uterine tissues were obtained from all groups for immunohistochemical analysis, and paraffin sections were obtained. Adipose tissue was used as a positive control.&#x0D; &#x0D; Results: In the Knt group, APLN protein expression was present especially in the uterine glands and stromal cells that are located close to the glands. The localization of APJ protein expression was also similar to APLN. In the GK group, APLN and APJ expression levels were similar to the Knt group. However, when the localization of APJ was evaluated, it has been observed that APJ expression was intensely expressed especially in the perimetrial glands of the uterus, which was not present in the Knt group. In the P+H group, APLN expression significantly increased in the endometrial glands and stromal cells compared to the Knt and GK groups (p

Measurement of glomerular filtration rate reveals that subcapsular injection of shear-thinning hyaluronic acid hydrogels does not impair kidney function in mice.

The continued development of minimally invasive therapeutic implants, such as injectable hydrogels, necessitates the concurrent advancement of methods to best assess their biocompatibility via functional outcomes in vivo. Biomaterial implants have been studied to treat kidney disease; however, assessment of biocompatibility has been limited to biomarker and histological assessments. Techniques now exist to measure kidney function serially in vivo in murine studies via transcutaneous measurements of glomerular filtration rate (tGFR). In this study, adult male and female wild‐type BalbC mice underwent right unilateral nephrectomy. The remaining solitary left kidney was allowed 4 weeks to recover via compensatory hypertrophy, after which subcapsular injection of either saline or shear‐thinning hyaluronic acid hydrogel was performed. Serial tGFR measurements before and after treatment were used to assess the effect of hydrogel injection on kidney filtration. Urine and serum biomarkers of kidney function, and kidney histology were also quantified. Hydrogel injection did not affect kidney function, as assessed by tGFR. Results were in agreement with standard metrics of serum and urine biomarkers of injury as well as histological assessment of inflammation. The model developed provides a direct functional assessment of implant compatibility for the treatment of kidney disease and impact on kidney function.

Can UVA radiation have a germicidal effect?

This study examines the efficiency of UVA radiation in the eradication of murine mites. The experiment consisted of the radiation of female BALBc mice in a solarium by UVA rays, up to total dosages of 7800 J/cm2and 12 500 J/cm2. A total of 88 animals were divided into four experimental and three control groups. Animals from the experimental and two control groups were shaved, and animals in two experimental groups were treated by a photoprotective cream half an hour before exposure. The animals in the third control group were not shaven. The presence of mites was detected in histological preparations of most of the animals in the control groups. When compared to the experimental groups of animals exposed to radiation up to 7800 J/cm2, the presence of mites was significantly more frequent in the control group of shaven animals (p = 0.013) and unshaven animals (p &lt; 0.001). When compared to the experimental groups exposed to radiation up to 12 500 J/cm2, the presence of mites was significantly more frequent in the control group of shaven animals (p &lt; 0.001) and unshaven animals (p &lt; 0.001). Median value of mites was statistically much higher in the control, unshaven group of animals in comparison to the untreated group of animals exposed to radiation up to 7800 J/cm2(p = 0.006). Median value of mites between untreated group of animals exposed to radiation up to 7800 J/cm2and their shaven control group was near a statistically significant difference (p = 0.056). By comparing the number of mites between the control groups of animals, no presence of statistically significant difference (p = 0.901) was confirmed. UVA radiation led to the effective eradication of murine mites.

The structural characteristics of photoaging in mice caused by the effects of ultraviolet A radiation.

Due to its deep penetration into the dermis, ultraviolet A (UVA) radiation is considered a primary factor in skin photoaging. The aim of this study is to use a qualitative and quantitative analysis to determine the structural parameters of skin photoaging in mice exposed to UVA radiation, with or without the application of a photoprotective cream. The experiment consisted of the radiation of female BALBc mice in a solarium by UVA rays, up to total dosages of 7800 J/cm² and 12500J/cm². A total of 78 animals were divided into 4 experimental and 2 control groups. All animals were shaved and the animals in 2 experimental groups were treated with a photoprotective cream half an hour before exposure. The samples of the treated skin were stained with Hematoxylin-eosin and Van-Gieson staining methods. All measurements, except for the presence of dyskeratosis, were taken using ImageJ 150i software. In the study, the signs of skin photoaging were more evident in untreated groups of animals. Dysceratosis was more frequent in both of the untreated groups of animals (p=0.004) and (p=0.003). The lowest values of epidermal thickness (13.8± 2.6μm and 12.7±2.3μm) were present in both of the untreated groups of animals (p<0.001) and (p<0.001). The highest values of stratum corneum thickness (34.3±8.5μm) were observed in the untreated, shorter radiated group of animals (p<0.001) which was irradiated for the shortest period of time. Beside the control groups, the highest length of dermo-epidermal junction (DEJ) was recorded in the group of treated, longer radiated animals (1467.6±94.6μm) (p=0.373). The lowest values of dermal thickness (115.9±10.5μm and 134.8±21.8μm) and volumetric density of the collagen fibers (31.92±3.19% and 29.40±4.54%) were present in both untreated groups of animals (p<0.001), (p<0.001), (p=0.035). Skin photoaging was most pronounced in the groups of animals irradiated without the application of photoprotective cream.

Calcium Fructoborate Prevents Skin Cancer Development in Balb-c Mice.

Tumor microenvironment, genetic, and non-genetic factors are responsible for the atypical metabolic feature of cancer cells. Aberrant activity of PI3K/Akt pathway, increased glycolytic flux, and decreased intracellular pH gradient are the leading causes of this feature. Calcium Fructoborate (CaFB), a sugar-borate ester, has major benefits for human health. The aim of this study was to explore the implication of CaFB on experimentally induced skin cancer in vivo. According to the treatment, 92 female Balb-c mice are divided into six groups: control, CaFB (3mg/kg/day), 7,12-Dimethylbenz(a)anthracene (DMBA)+12-O-tetradecanoylphorbol-13-acetate (TPA) (97.5nmol DMBA, 6.5nmol TPA), T1: CaFB+DMBA+TPA (3mg/kg/day CaFB together with DMBA), T2: DMBA+CaFB+TPA (3mg/kg/day CaFB together with TPA), T3: DMBA+TPA+CaFB (3mg/kg/day CaFB after tumor formation). Topical DMBA and TPA application resulted in a significant increase in the protein levels, immunoreactivity, and mRNA expression of HRAS, HIF1α, Akt, and PTEN (p < 0.05). Moreover, an increase in the number of TUNEL-positive cells was observed in DMBA-TPA group compared with the control group (p < 0.05). CaFB application reduced the protein levels, immunoreactivity, and mRNA expressions of HRAS, HIF1α, Akt, and PTEN and also decreased the number of TUNEL-positive cells. Recent evidence obtained from our study validated that CaFB treatment may have skin cancer-preventing effect.

Low Protein/low Methionine/high Carbohydrate Diets Induce Hyperphagia, Increase Energy Expenditure and FGF21, but Modestly Affect Adiposity in Female BalbC Mice (OR09-01-19)

ObjectivesLow-protein diets are reported to induce hyperphagia to fulfil protein needs but at the expense of energy balance with a risk to gain in adiposity. However, different studies did not show body fat gain because an increased energy expenditure partly compensated the increase in energy intake and prevents the gain in adiposity. The present study evaluated in mice the consequence of protein restricted diets combined with protein quality (milk protein versus soy protein with slight methionine deficiency) on energy balance and adiposity and the role of FGF21 in the response to the protein restricted diets. MethodsThe study investigated in female BalbC mice the behavioural, metabolic and phenotypic responses to 8 weeks feeding a very low (3% energy - P3), moderately low (6% - P6) or adequate (20% - P20) dietary protein diet and evaluated if methionine scarcity, using soy protein (S) vs casein (C), affected these responses. Food intake, body weight, adiposity (assessed by DEXA), were measured throughout the study and body composition determined at the end of the study. Plasma, liver, muscle, adipose tissue and hypothalamus samples were collected for nutrient, hormones and gene expression measurements. ResultsDecreasing dietary casein from 20% to 3% increased energy intake, slightly increased adiposity, and this was exacerbated with methionine-deficient soy protein (figure 1). Lean body mass was reduced in 3% casein fed mice but preserved in all 6% fed mice. The effect on fat mass was limited because energy expenditure was also increased (figure 2). In plasma, low protein diets decreased IGF-1 and increased FGF21 that was related to protein level, protein to carbohydrate ratio and methionine content in the diet (figure 3). Insulin response to an oral glucose test was reduced in soy and low-protein fed mice. Low-protein diets did not affect Ucp1 but increased Fgf21 in brown adipose tissue and Fgf21, Fas, and Cd36 in the liver. In the hypothalamus, Npy was increased and Pomc was decreased only in 3% casein fed mice. ConclusionsReducing dietary protein and protein quality increases both energy intake and energy expenditure resulting only in slight increase in adiposity. In this process FGF21 is probably a signal that responds to a combination of protein restriction and carbohydrate content of the diet. Funding SourcesInstitut National de la Recherche Agronomique, AgroParisTech. Supporting Tables, Images and/or Graphs▪▪▪

Antimicrobial photodynamic therapy as a new approach for the treatment of vulvovaginal candidiasis: preliminary results.

In this work, we present the efficacy of photodynamic therapy against yeast cells in an animal model. We tested two photosensitizers, methylene blue and protoporphyrin IX. Thirty-seven female BALB-c mice with a body mass of 20-25g were used. To achieve persistent vaginitis, estrus was induced by subcutaneous injection of 0.1mg/mL estradiol valerate applied weekly. Three days after pseudo-estrus, intravaginal inoculation with Candida albicans was performed. Mice were anesthetized with ketamine (80mg/kg) and xylazine (10mg/kg) by intraperitoneal injection before inoculation, and antimicrobial photodynamic therapy (aPDT) was performed 5days after fungal inoculation. Two photosensitizers were tested, methylene blue (MB; 100μM) and protoporphyrin IX (PpNetNI; 10μM). Two custom-made LEDs emitting light at 660 and 630nm at approximately 800mW each were used for irradiation. The aPDT treatment reduced the fungal colony-forming units (CFUs) by one order of magnitude for the MB (p = 0.020) and PpNetNI (p = 0.018) photosensitizers. Seven days after the treatment, there were significantly fewer CFUs compared to the control group (p = 0.041 and p = 0.035 for MB and PpNetNI, respectively), but this was not increased compared to the initial number immediately after aPDT. Using aPDT as a therapeutic option to decrease fungal infection in a vaginal candidiasis model resulted in a significant reduction in the C. albicans population. Both photosensitizers were effective for preventing reinfection within 7days. The aPDT also had no effect on the vaginal mucosa at the ultrastructural level. In addition to the fungicide effect, we observed reduced swelling and lack of the formation of abscesses, microabscesses coating the cornified epithelial layer, and the accumulation of neutrophils in the submucosa.

Effects of pentoxifylline on the histological and ultra-structural features of vitrified mouse ovarian tissue: An experimental study

Vitrification is a process that can be used to preserve gonads in the healthy and natural status. Oxidative stress is one of the disadvantages of vitrification. Pentoxifylline (PTX) is an antioxidant that can reduce reactive oxidative stress effects. We aimed to investigate the effects of PTX on histological and ultra-structural features of vitrified and non-vitrified mouse ovarian tissue. Twenty-five adult female Balb-C mice were randomly and equally divided into control group: the ovaries did not receive any treatment; experimental 1 and 2: the vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin without and with PTX, respectively, for 30 min; sham 1 and 2: the non-vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin and were incubated without and with PTX, respectively for 30 min. The right and left ovaries in all of the groups were evaluated using light and transmission electron microscopy, respectively. The histological and ultra-structural features of vitrified ovaries were seriously damaged. There was non-uniformed germinal epithelium and tunica albuginea, degenerated granulosa cells and stromal cells, puffy basement membrane and irregular thickness of zona pellucida, as well as a pyknotic nucleus and bubbly and segmented ooplasmic in the follicles. Also, ovarian tissues were damaged by the PTX in the non-vitrified ovaries. Vitrification can damage the histological and ultra-structural features of the ovary in mouse models. PTX as an antioxidant, with concentration of 1.8 mM could not prevent and restore these damages and had no adequate effects on the vitrified ovarian tissues.

Gut microbiota in experimental murine model of Graves\u2019 orbitopathy established in different environments may modulate clinical presentation of disease

BackgroundVariation in induced models of autoimmunity has been attributed to the housing environment and its effect on the gut microbiota. In Graves’ disease (GD), autoantibodies to the thyrotropin receptor (TSHR) cause autoimmune hyperthyroidism. Many GD patients develop Graves’ orbitopathy or ophthalmopathy (GO) characterized by orbital tissue remodeling including adipogenesis. Murine models of GD/GO would help delineate pathogenetic mechanisms, and although several have been reported, most lack reproducibility. A model comprising immunization of female BALBc mice with a TSHR expression plasmid using in vivo electroporation was reproduced in two independent laboratories. Similar orbital disease was induced in both centers, but differences were apparent (e.g., hyperthyroidism in Center 1 but not Center 2). We hypothesized a role for the gut microbiota influencing the outcome and reproducibility of induced GO.ResultsWe combined metataxonomics (16S rRNA gene sequencing) and traditional microbial culture of the intestinal contents from the GO murine model, to analyze the gut microbiota in the two centers. We observed significant differences in alpha and beta diversity and in the taxonomic profiles, e.g., operational taxonomic units (OTUs) from the genus Lactobacillus were more abundant in Center 2, and Bacteroides and Bifidobacterium counts were more abundant in Center 1 where we also observed a negative correlation between the OTUs of the genus Intestinimonas and TSHR autoantibodies. Traditional microbiology largely confirmed the metataxonomics data and indicated significantly higher yeast counts in Center 1 TSHR-immunized mice. We also compared the gut microbiota between immunization groups within Center 2, comprising the TSHR- or βgal control-immunized mice and naïve untreated mice. We observed a shift of the TSHR-immunized mice bacterial communities described by the beta diversity weighted Unifrac. Furthermore, we observed a significant positive correlation between the presence of Firmicutes and orbital-adipogenesis specifically in TSHR-immunized mice.ConclusionsThe significant differences observed in microbiota composition from BALBc mice undergoing the same immunization protocol in comparable specific-pathogen-free (SPF) units in different centers support a role for the gut microbiota in modulating the induced response. The gut microbiota might also contribute to the heterogeneity of induced response since we report potential disease-associated microbial taxonomies and correlation with ocular disease.

Attenuation of Vascular Contractility in Metastatic Breast Cancer Mice

Breast cancer (BC) is the one of crucial health burden among women worldwide. Even though the alteration of vascular function were observed by few researchers in metastatic BC, the changes in vascular reactivity and its underlying mechanism were not fully understood. Here, we investigate the physiological changes of vascular function from isolated mesenteric arteries in metastatic breast cancer mice model. In this study, 105 of 4T1 murine BC cells were injected into the subcutaneous or tail vein of female Balb‐c mice aged 8 weeks old. Mice were randomly divided into the following group: 1) control; n=10, 2) non‐metastatic model (4T1; n=10), 3) tumor‐bearing model (4T1M; n=10), and 4) tail vein injection model (4T1ML; n=10). Mesenteric arteries were removed after 4 weeks of tail vein injection group and 5 weeks of tumor‐bearing groups. Isometric tension was investigated in response to various vasoconstrictors such as phenylephrine (PhE), Serotonin (5‐HT), Angiotensin II (ANG II) and high potassium chloride (HK). Our results showed that PhE‐induced vasoconstriction was no significant differences in the non‐metastatic group compared to control. However, the attenuated vasoconstriction in response to PhE, 5‐HT, and High K was found in metastatic cancer groups. Application of L‐type voltage‐gated calcium channel inhibitor (Nifedipine) induced vascular relaxation with PhE‐induced contraction in each group, but there were no significant differences among groups. Interestingly, PhE‐induced contraction in metastatic groups was not inhibited by Rho kinase inhibitor (Y‐27632), but not in control. From above results, it is suggested that decreased Rho kinase activity might be attributable to attenuated vasoconstriction in mesenteric artery from advanced breast cancer mice.Support or Funding InformationThis study was supported by a National Research Foundation of Korea grant funded by Ministry of Science, ICT, &amp; Future Planning (2015R1C1A1A01054038).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 Effects of Photobiomodulation of 808 nm Diode Laser Therapy at Higher Fluence on the in Vitro Osteogenic Differentiation of Bone Marrow Stromal Cells

The literature has supported the concept of mesenchymal stromal cells (MSCs) in bone regeneration as one of the most important applications in oro-maxillofacial reconstructions. However, the fate of the transplanted cells and their effects on the clinical outcome is still uncertain. Photobiomodulation (PBM) plays an important role in the acceleration of tissue regeneration and potential repair. The aim of this in vitro study is to evaluate the effectiveness of PBM with 808 nm diode laser therapy, using a flat-top hand-piece delivery system at a higher-fluence (64 J/cm2) irradiation (1 W, continuous-wave) on bone marrow stromal cells (BMSCs). The BMSCs of 3 old female Balb-c mice were analyzed. The cells were divided into two groups: irradiated group and control group. In the former the cells were irradiated every 24 h during 0 day (T0), 5 (T1), 10 (T2), and 15 (T3) days, whereas the control group was non-irradiated. The results have shown that the 64 J/cm2 laser irradiation has increased the Runt-related transcription factor 2 (Runx2). Runx2 is the most important early marker of osteoblast differentiation. The higher-fluence suppressed the synthesis of adipogenic transcription factor (PPARγ), the pivotal transcription factor in adipogenic differentiation. Also, the osteogenic markers such as Osterix (Osx) and alkaline phosphatase (ALP) were upregulated with an increase in the matrix mineralization. Furthermore, western blotting data demonstrated that the laser therapy has induced a statistically valid increase in the synthesis of transforming growth factor β1 (TGF-β1) but had no effects on the tumor necrosis factor α (TNFα) production. The data has statistically validated the down-regulation of the important pro-inflammatory cytokines such as interleukin IL-6, and IL-17 after 808 nm PBM exposition. An increase in anti-inflammatory cytokines such as IL-1rα and IL-10 was observed. These in vitro studies provide for first time the initial proof that the PBM of the 808 nm diode laser therapy with flat-top hand-piece delivery system at a higher-fluence irradiation of 64 J/cm2 (1 W/cm2) can modulate BMSCs differentiation in enhancing osteogenesis.

Effects of epigallocatechin-3-gallate (EGCG) on a scleroderma model of fibrosis

Abstract Objective The aim of the present study was to evaluate the potential protective effects of epigallocatechin-3-gallate (EGCG) on fibrosis in bleomycin induced scleroderma model. Materials and methods Thirty-two healthy female Balb-c mice with the average body weight of 22±5 g were used in this study. The mice were randomly divided into four groups as control (n=8), Bleomycin (n=8), Bleomycin+EGCG (n=8) and EGCG (n=8). Skin tissue samples were collected to quantify matrix metalloproteinases (MMP-1, MMP-8, MMP-13), p-SMAD 2/3 and SMAD 2/3 in protein homogenates by western blotting. TGF-β1 expression was determined by real-time PCR. Immunohistopathological and histopathological examinations of skin tissues were also done. Results When measured with Masson Trichrome, EGCG treatment was found to decrease fibrosis in connective tissue compared to the BLM injected control. EGCG was decreased dermal fibrosis. Bleomycin+EGCG group showed a significant reduction in fibrosis at the dermal surface area using hematoxylin measurements compared with the BLM group. MMP-1, MMP-8 protein levels were increased and p-SMAD 2/3 protein level was decreased. TGF-β mRNA expression was decreased in the EGCG+BLM group compared with the BLM group. Conclusion These results suggest an antifibrotic role for EGCG.

Exposure to Low-Dose X-Ray Radiation Alters Bone Progenitor Cells and Bone Microarchitecture.

Exposure to high-dose ionizing radiation during medical treatment exerts well-documented deleterious effects on bone health, reducing bone density and contributing to bone growth retardation in young patients and spontaneous fracture in postmenopausal women. However, the majority of human radiation exposures occur in a much lower dose range than that used in the radiation oncology clinic. Furthermore, very few studies have examined the effects of low-dose ionizing radiation on bone integrity and results have been inconsistent. In this study, mice were irradiated with a total-body dose of 0.17, 0.5 or 1 Gy to quantify the early (day 3 postirradiation) and delayed (day 21 postirradiation) effects of radiation on bone microarchitecture and bone marrow stromal cells (BMSCs). Female BALBc mice (4 months old) were divided into four groups: irradiated (0.17, 0.5 and 1 Gy) and sham-irradiated controls (0 Gy). Micro-computed tomography analysis of distal femur trabecular bone from animals at day 21 after exposure to 1 Gy of X-ray radiation revealed a 21% smaller bone volume (BV/TV), 22% decrease in trabecular numbers (Tb.N) and 9% greater trabecular separation (Tb.Sp) compared to sham-irradiated controls (P < 0.05). We evaluated the differentiation capacity of bone marrow stromal cells harvested at days 3 and 21 postirradiation into osteoblast and adipocyte cells. Osteoblast and adipocyte differentiation was decreased when cells were harvested at day 3 postirradiation but enhanced in cells isolated at day 21 postirradiation, suggesting a compensatory recovery process. Osteoclast differentiation was increased in 1 Gy irradiated BMSCs harvested at day 3 postirradiation, but not in those harvested at day 21 postirradiation, compared to controls. This study provides evidence of an early, radiation-induced decrease in osteoblast activity and numbers, as well as a later recovery effect after exposure to 1 Gy of X-rays, whereas osteoclastogenesis was enhanced. A better understanding of the effects of radiation on osteoprogenitor cell populations could lead to more effective therapeutic interventions that protect bone integrity for individuals exposed to low-dose ionizing radiation.