Effect of ascorbic acid and epidermal growth factor in a rat tibia defect

Purpose: Bone repair aims to restore the anatomical, biomechanical, and functional integrity of the affected structure. Here we study the effects of ascorbic acid (AA) and epidermal growth factor (EGF) applied in a single dose and in combination on the repair of a noncritical bone defect model. Methods: Twenty-four rats were divided into four groups: an intact G-1 control group, and three groups that underwent a noncritical bone defect in the right tibia: G-2 treated with AA, G-3 treated with EGF, and G-4 treated with AA in combination with EGF. After 21 days of treatment, rats were sacrificed, the tibias were dissected and a destructive biomechanical analysis of three-point flexion test was performed in a universal testing machine; the values of stiffness, resistance, maximum energy, and energy at maximum load were statistically compared. Results: G-3 and G-4 recovered the biomechanical properties of strength and stiffness of an intact tibia 3 weeks after their application. Not so the energy and energy at maximum load. For G-2, only the stiffness of an intact tibia was recovered. Conclusion: EGF and AA-EGF applied to a noncritical bone defect in the rat tibia favors the recovery of bone resistance and stiffness.

The effect of olsalazine of chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt in BALB/c mice

Purpose: To explore effect and mechanism of olsalazine of Chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt (DSS) in BALB/c mice. Methods: The mouse model of ulcerative colitis was induced by free drinking of 3% (w/v) DSS aqueous solution for seven days. The mice were treated with olsalazine (0.6 g·kg-1) of Chinese generic drugs. The therapeutic effect of olsalazine on ulcerative colitis mice was evaluated by measuring disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS), and detected the expression levels of interleukin (IL)-2, IL-10, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-1ß in serum and IL-7, IL-17, IL-22, epidermal growth factor (EGF), transforming growth factor ß1 (TGF-ß1) in colonic homogenate of mice. Results: Olsalazine significantly increased the contents of IL-2, IL-10, IL-22, TGF and EGF in ulcerative colitis rats, and significantly decreased the scores of DAI, CMDI, HS and the contents in IL-7, IL-17, TNF-α, IL-1ß and IFN-γ when compared with the model group. It improved the degree of colonic lesion in ulcerative colitis mice. Conclusions: It was suggested that olsalazine has a therapeutic effect on ulcerative colitis induced by DSS in mice, and the mechanism may be related to the increase of IL-2, IL-10, IL-22, TGF, and EGF and the decrease of the expression of IL-7, IL-17, TNF-α, IL-1ß, and IFN-γ.

The ethanol extract of Periplaneta Americana L. improves ulcerative colitis induced by a combination of chronic stress and TNBS in rats

Purpose: To investigate the effects of Periplaneta americana L. on ulcerative colitis (UC) induced by a combination of chronic stress (CS) and 2,4,6-trinitrobenzene sulfonic acid enema (TNBS) in rats. Methods: The experiment UC model with CS was established in rats by a combination of chronic restraint stress, excess failure, improper, and TNBS. The body weight, disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS) and pro-inflammatory mediators were measured. The content of corticotropin-releasing hormone (CRH) in hypothalamus or adrenocorticotropic hormone (ACTH) and corticosteroids (CORT) in plasma were evaluated by enzyme-linked immunosorbent assay. The proportion of T lymphocyte subsets was detected by flow cytometry, and gut microbiota was detected by 16S rDNA amplicon sequencing. Results: Weight loss, DAI, CMDI, HS and proinflammatory mediators were reversed in rats by P. americana L. treatment after UC with CS. Increased epidermal growth factor (EGF) was observed in P. americana L. groups. In addition, P. americana L. could reduce the content of CRH and ACTH and regulate the ratio of CD3+, CD3+CD8+ and CD3+CD4+CD25+/CD4+ in spleen. Comparably, P. americana L. changes composition of gut microbiota. Conclusions: The ethanol extract of Periplaneta Americana L. improves UC induced by a combination of CS and TNBS in rats.

The inhibitory and anticancer properties of Annona squamosa L. seed extracts / Propriedades inibitórias e anticancerígenas de extratos de sementes de Annona squamosa L

Although Annona squamosa Linn. (Annonaceae) has been used in traditional medicine and is known to have several pharmacological properties, its impact on EGFR kinase has not been fully investigated. An assay (biochemical) was used to govern the potential of different A. squamosa seed extracts to scavenge free radicals in petroleum ether, acetone, ethanol, and methanol. We also tested A. squamosa leaf extracts for their ability to inhibit the growth of HEK 293, MCF7, and HepG2 cell lines. The PSE, ASE, ESE, and MSE all contained anti-cancer substances like anethole, cyclopentane, 1,1,3-trimethyl, and phosphonate oxide tributyl, according to phytochemical analysis. ESE extracts from A. squamosa seeds have been selected based on free radical generation probabilities, cytotoxicity studies, and phytochemical analysis. Subsequent insilico studies have been conducted, and the results have shown that interactions between compounds present in ESE extracts and the EGFR kinase are what give these compounds their inhibitory effects. Preliminary phytochemical and pharmacological activities were studied and reported. A. squamosa ESE extracts inhibited the growth of MCF7 cells, and a pharmacokinetic study showed that the compounds anethole, cyclopentane, 1,1,3-trimethyl, and phosphonium oxide tributyl had few undesirable side effects. These substances can be used to both prevent and treat cancer diseases.

Embora a Annona squamosa Linn. (Annonaceae) tenha sido utilizada na medicina tradicional e seja conhecida por diversas propriedades farmacológicas, seu impacto na EGFR quinase ainda não foi totalmente investigado. Um ensaio bioquímico foi utilizado para controlar o potencial de diferentes extratos de sementes de A. squamosa para eliminar radicais livres em éter de petróleo, acetona, etanol e metanol. Extratos de folhas de A. squamosa também foram analisados em relação à sua capacidade de inibir o crescimento de linhagens celulares HEK 293, MCF7 e HepG2. O PSE, ASE, ESE e MSE continham substâncias anticancerígenas como anetol, ciclopentano, 1,1,3-trimetil e óxido de fosfonato tributil, de acordo com a análise fitoquímica. Extratos de ESE de sementes de A. squamosa foram selecionados com base em probabilidades de geração de radicais livres, estudos de citotoxicidade e análise fitoquímica. Estudos in silico subsequentes foram realizados e os resultados mostraram que as interações entre os compostos presentes nos extratos de ESE e a EGFR quinase são o que confere a esses compostos seus efeitos inibitórios. As atividades fitoquímicas e farmacológicas preliminares foram estudadas e relatadas. Os extratos de ESSE de A. squamosa inibiram o crescimento de células MCF7, e um estudo farmacocinético mostrou que os compostos anetol, ciclopentano, 1,1,3-trimetil e óxido de fosfônio tributil tiveram poucos efeitos colaterais indesejáveis. Essas substâncias podem ser usadas para prevenir e tratar doenças cancerígenas.

Prevalence and level of infection by endoparasites in different categories of sheep in the Western Amazon / Prevalência e nível de infecção por endoparasitas em diferentes categorias de ovinos na Amazônia Ocidental

This study aimed to understand the epidemiology of the main gastrointestinal endoparasites affecting sheep raised in the municipality of Sena Madureira, Acre, Brazil. A total of 178 fecal samples were collected from sheep raised in an extensive system of 10 farms. The samples were grouped into lambs, ewe lambs, lambing ewes, ewes, and rams for laboratory analysis by the flotation technique for counting eggs per gram of feces (EGF), oocysts of Eimeria sp., and presence of eggs of Moniezia sp. The analyzed variables consisted of the prevalence of nematodes, cestodes, and coccidia and intensity of strongylid and coccidium infection by quantification (mean ± SE) of eggs and oocysts. Prevalence data were compared by the chisquare test and intensity of infection (mean ± SE) by the Scott-Knott test (SAEG 9.1), both with P < 0.05. The overall prevalence was 77.6%, reaching 64.15% for strongylids, 36.2% for coccidia, and 8.81% for cestodes (Moniezia sp.). Ewes had the lowest prevalence (52.5%), while lambs had a prevalence of 95.5%, not differing from the others (P < 0.05). The intensity of infection by nematodes showed that lambs had the highest EGF compared to ewes, with values of 1297±270 and 232±79, respectively. The categories lambing ewes, ewes, and rams presented an average EGF below what is indicated for treatment with drugs. The highest intensity of infection was observed for ewe lambs (5859±3648), exceeding acceptable rates. Sheep raised in Sena Madureira has a high prevalence for verminosis by strongylids and a low prevalence for coccidiosis and cestodiasis. Young animal categories present a high health risk for endoparasitoses, and management measures that collaborate with the prophylaxis and control of these diseases should be adopted.(AU)

Objetivou-se conhecer a epidemiologia dos principais endoparasitas gastrointestinais que acometem ovinos criados no município de Sena Madureira-Acre. Foram coletadas 178 amostras fecais de ovinos criados em sistema extensivo de 10 propriedades rurais. As amostras foram agrupadas: cordeiros, marrãs, ovelhas paridas, ovelhas solteiras e reprodutores para análise laboratorial pela técnica de flutuação para a contagem de ovos por grama de fezes (OPG), oocistos de Eimeria sp. e presença de ovos de Moniezia sp. As variáveis analisadas foram: prevalência dos nematódeos, cestódeos e coccídios; intensidade de infecção por estrongilídeos e coccídios pela quantificação (média±EP) de ovos e oocistos. Os dados de prevalência foram comparados pelo teste Qui-quadrado, intensidade da infecção (média±EP) pelo teste Scott-Knott (SAEG 9.1), ambos com P < 0,05. A prevalência geral encontrada foi de 77,6%, para estrongilídeos foi 64,15%, coccídios 36,2% e cestódeos (Moniezia sp.) 8,81%. As ovelhas solteiras apresentaram a menor prevalência (52,5%), já os cordeiros apresentaram 95,5%, não diferindo das demais (P < 0,05). Quanto a intensidade da infecção por verminoses por nematódeos, os cordeiros apresentaram OPG mais elevado versus ovelhas solteiras 1297±270 vs. 232±79, respectivamente. As categorias ovelhas paridas, solteiras e reprodutores apresentaram média de OPG abaixo do que é indicado para tratamento com fármacos. A intensidade de infecção mais alta foi das marrãs (5859±3648), superando taxas aceitáveis. Ovinos criados em Sena Madureira apresentam alta prevalência para verminose por estrongilídeos, baixas prevalências para coccidiose e cestodioses. Categorias animais jovens apresentam elevado risco sanitário para as endoparasitoses, devendo-se adotar medidas de manejo que colaborem com a profilaxia e controle dessas enfermidades.(AU)

Changes in somatic cell count, composition and cytokine levels in milk from cows with mastitis due to mixed infections

Mastitis is a mammary gland inflammation that is very common worldwide, mostly caused by bacteria, and causes enormous economic losses. Many microorganisms cause this disease. The most common causes of mastitis by these microorganisms are Staphylococcus aureus (S. aureus), Escherichia coli (E. coli) and Streptococcus agalactiae (S. agalactiae). The anti-inflammatory properties of transforming growth factor (TGF)-ß include: 1) limiting interferon (IFN)-γ production; 2) increasing the expression of the interleukine (IL)-1 receptor antagonist; 3) inhibiting macrophage production of chemokines, pro-inflammatory cytokines, nitric oxide, and reactive oxygen intermediates; and 4) increasing macrophage clearance of bacterial debris and damaged parenchymal cells. It is stated that cytokines and milk composition change in case of mastitis. In this study, it was aimed to reveal the changes in milk TGF-ß1 and Tumor necrosis factor (TNF)-α concentrations and milk composition in mixed infections caused by three pathogens causing mastitis. In this study, milk samples from 90 cows were divided into 5 groups. Tumor necrosis factor (TNF)-α and TGF-ß1 concentrations and milk composition were determined in these milk samples. The California Mastitis Test (CMT) was applied to the cows included in the study and scoring was done. According to the CMT results of the milk samples taken, CMT(-) cows were included in group 1 (n = 22). Those with the CMT(+) were sent to the microbiology laboratory for analysis within 2 h. After the bacteria was determined, combination groupings were formed. Group 2 (n = 17), in which S. aureus and E. coli grew together, group 3 (n = 21), in which S. aureus and S. agalactiae grew together, group 4 (n = 8), in which S. agalactiae and E. coli grew together in milk samples, and milk samples without any bacterial growth in CMT (+) formed group 5 (n = 22), respectively. Somatic cell count was measured with the DeLaval Cell Counter® (Cell Counter DCC) device. Mineral matter, fat, protein, lactose, electrical conductivity and specific gravity were measured in milk samples using Lactoscan Milk Analyzer (Milkotronic/EUROPE). Milk samples were then stored at -80°C to measure TGF-ß1 and TNF-α. Tumor necrosis factor-α and TGF-ß1 concentrations in milk samples were measured using ELISA kits (Sunred Biological Technology). Changes in milk TNF-α and TGF-ß1 concentration and milk composition were determined in milk samples with mastitis caused by mixed infection. The TNF-α concentration of group 4 was higher than the other groups. On the other hand, the highest concentration of TGF-ß1 was found in group 2. While the number of somatic cells in group 1 was lower than in groups 2, 3, and 4, there was no statistical difference between groups 1 and 5. The lowest milk fat ratio was found in group 1, and it was found to be statistically lower than groups 2, 3, and 4. While the rate of solid-non-fat of group 1 increased compared to groups 2 and 3, the highest protein ratio was found in groups 1 and 5. There was no difference between the 5 groups in terms of mineral matter ratios. While the specific gravity was highest in group 1, there was no statistical difference between the other 4 groups. Overall, it was concluded that there was an increase in TNF-α and TGF-ß1 concentrations and a change in milk composition in samples with bacterial growth.(AU)

Epidermal renewal after treatment of primary canine hypothyroidism using levothyroxine

Background: Primary canine hypothyroidism, an endocrine disorder that causes imbalances in the hypothalamus-pituitarythyroid axis, is a common cause of endocrine dermatoses, which frequently presents with opaque dry brittle hair. Tissue changes are also visible, such as myxedema, hyperkeratosis, epidermal atrophy, alopecia, and others. This paper describes the skin changes caused by primary hypothyroidism in a female dog before and after treatment with levothyroxine. Case: This case study involved a 7-year-old Dalmatian bitch with a history of weight gain and changes such as rough dry brittle hair. For about a year, the dog also had also presented symmetrical erythematous and alopecia skin lesions in the regions of the hind limbs, lower back and tail, progressive lethargy and fatigue after exercise. A hemogram showed mild normocytic and normochromic regenerative anemia, as well as mild leukopenia and neutropenia. Biochemical tests revealed increased levels of creatinine, total cholesterol, and triglycerides. Skin biopsy revealed the presence of comedones in the epidermis and acanthosis and trichilemmal keratinization. Hormonal tests revealed high TSH and low free T4 and total T4 levels. A cervical ultrasound scan showed changes in the thyroid glands, with hypoechoic parenchyma, in addition to increased size of the right lobe, suggesting thyroiditis. Based on these exams, the patient was diagnosed with primary hypothyroidism, and treatment with levothyroxine was instituted. Discussion: Around 90% of dogs with hypothyroidism show a decrease in total T4 and about 65-75% of them show an increase in TSH levels. In endocrinopathy, there is a decrease in the expression of T3 receptors in the keratinocytes and genes responsible for epidermal renewal. Also, molecules that affect epidermal differentiation bind to intracellular receptors belonging to the steroid/thyroid hormone superfamily. In this study, however, the application of the minimum levothyroxine dose restored the normal epidermal pattern for the species and the remaining parameters returned to normal. The hormone T3 acts in the differentiation of keratinocytes, a fact that, although not yet proven, is believed to be indirect and mediated by the epidermal growth factor or by the expression of the genes responsible for the renewal of the epidermis. Besides, some effects on the epidermis caused by deficiency of thyroid hormones may be due to secondary vitamin A deficiency, which is necessary for epithelial differentiation and binding epidermal growth factor (EGF) to the cell. Still, there is a theory that sex hormones also affect the differentiation of the epidermis, as studies show that deficiency causes effects similar to those caused by hypothyroidism. Some studies indicate the investigation of the relationship between secondary vitamin D deficiency and the increased cellularity of the epidermis of the spayed hypothyroid female rats. Additionally, the mechanism of the formation of hyperkeratosis in hypothyroidism is not yet elucidated. T3 is known to regulate keratin gene expression and perhaps also epidermal maturation, epithelial cycle, and normal keratin synthesis. The single treatment with levothyroxine in the female dog was effective in promoting the restoration of gene expression to T3 in keratinocytes.

To explore the therapeutic effect of Kangfuxiaomi shuan II on cervicitis in rats

ABSTRACT Purpose: The rat cervicitis model was established with 20% phenol glue to explore the therapeutic effect of Kangfuxiaomi shuan II on rat cervicitis and its mechanism. Methods: After modeling, the rats were treated with Shuangzuotai suppository (37.84 mg/kg), Kangfuxiaoyan shuan (205.6 mg/kg) and Kangfuxiaomi shuan II (40, 80, 160 mg/kg). The histopathological changes and injury degree of cervix in rats were evaluated by vulvar inflammation score and organ index. The therapeutic effect of Kangfuxiaomi shuan II on cervicitis was evaluated by detecting the levels of copper-protein (CP), C-reactive protein (CRP), Rat interleukin 6 (IL-6), superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and epidermal growth factor (EGF), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in cervical tissue. Results: Compared with the model group, the vulvar inflammation score and cervical index of rats in other groups decreased significantly (P<0.01). Kangfuxiaomi shuan II could significantly reduce the levels of CP, CRP, and MDA in serum of rats with cervicitis, and significantly increase the activity of SOD in serum of rats with cervicitis (P<0.01). The levels of EGF and iNOS in cervical tissue of rats also increased in different degrees, while the level of COX-2 decreased significantly (P<0.01), which significantly improved the pathological degree of vulvar inflammation in rats with cervicitis. Conclusions: Kangfuxiaomi shuan II has a certain therapeutic effect on cervicitis in rats, and its mechanism may be related to the regulation of inflammatory cytokine network and immunity.

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.(AU)

Osteopontin expression and its relationship with prognostic biomarkers in canine mammary carcinomas

ABSTRACT: Osteopontin is a glycophosphoprotein implicated in different physiologic and pathologic processes and is known to be involved in progression and metastasis of various cancers in humans, but this relation is still little explored in the veterinary. The aim was to evaluate the expression of osteopontin in canine mammary carcinomas and its relation with well-established canine mammary tumor biomarkers. For that, expression of OPN, EGFR, HER2, and c-Kit were evaluated along with Ki67 rate in 43 mammary carcinomas. Osteopontin was demonstrated to be expressed by neoplastic epithelial cells in all carcinomas as well as in stromal cells from the tumor microenvironment. Relation between high osteopontin expression and EGFR positivity (P 0.001) and HER2 overexpression (P=0.012) was demonstrated. In conclusion, high OPN expression seems to be related to poor prognosis and MAPK pathway activation, given the association with EGFR and HER2, members of the MAPK signaling pathway.

RESUMO: A osteopontina é uma glicofosfoproteina implicada em diferentes processos fisiológicos e patológicos, sendo conhecida por estar envolvida na progressão e metástase de vários cânceres nos humanos, no entanto, essa relação é ainda pouco explorada na veterinária. O objetivo deste trabalho foi avaliar a expressão da osteopontina nos carcinomas mamários caninos e sua relação com biomarcadores bem estabelecidos para esta neoplasia. Para isto, foi avaliada a expressão de OPN, EGRH, HER2 e c-Kit juntamente com a taxa de Ki67 em 43 carcinomas mamários. A osteopontina foi expressa pelas células epiteliais neoplásicas em todos os carcinomas, assim como, nas células estromais do microambiente tumoral. Foi demonstrada uma relação entre uma alta expressão de osteopontina e positividade para EGFR (P 0.001) e superexpressão de HER2 (P=0.012). Em conclusão, alta expressão de OPN parece estar relacionada com mau prognóstico e ativação da via MAPK, devido a sua associação com EGRF e HER2, os quais são membros desta via de sinalização.

Periplaneta americana extract promotes intestinal mucosa repair of ulcerative colitis in rat

Purpose: To investigate the mechanism of Periplaneta americana extract promoting intestinal mucosal repair of OXZ-induced colitis in rat. Methods: All experiments used an equal number of male and female SD rats (n=48). We injected OXZ into the colon to induce UC rat model. To determine the optimal concentration of P. Americana's extract (PA-40), it was classified into low (L), medium (M), and high (H) doses. After OXZ treatment, each drug was administered by enema for 7 consecutive days. Rats were divided into the following 6 groups: (1) Saline treatment group (NC), (2) OXZ treatment UC model group (MC), (3) OXZ + budesonide group (BUN), (4) OXZ + PA-40 L group, (5) OXZ + PA-40 M group, (6) OXZ + PA-40 H group. Disease activity index (DAI) scores, colon length, histopathological score, serum cytokine level (IL-4, IL-10, iNOS, tNOS), and amount of MPO, EGF, IL-13 in colonic mucosa were measured. Results: PA treatment had a significant healing effect on the OXZ-colitis model and significantly reduced the lesioned area, especially in the PA-40H groups. PA treatment did not alter the expression of IL-10 and MPO level, but increased EGF (epidermal growth factor) and decrease IL-13 in the colonic tissue. PA inhibited the rise of NOSs (nitric oxide synthase) and decreased the serum IL-4 level. Conclusions: The data suggest that Periplaneta americana extract may be a potential compound for the treatment of colonic lesions. The mechanism may be related to inhibiting the secretion of IL-13 and promoting the formation of EGF.(AU)

Osteopontin expression and its relationship with prognostic biomarkers in canine mammary carcinomas / Expressão de osteopontina e sua relação com biomarcadores prognóstico nos carcinomas mamários caninos

Osteopontin is a glycophosphoprotein implicated in different physiologic and pathologic processes and is known to be involved in progression and metastasis of various cancers in humans, but this relation is still little explored in the veterinary. The aim was to evaluate the expression of osteopontin in canine mammary carcinomas and its relation with well-established canine mammary tumor biomarkers. For that, expression of OPN, EGFR, HER2, and c-Kit were evaluated along with Ki67 rate in 43 mammary carcinomas. Osteopontin was demonstrated to be expressed by neoplastic epithelial cells in all carcinomas as well as in stromal cells from the tumor microenvironment. Relation between high osteopontin expression and EGFR positivity (P<0.001) and HER2 overexpression (P=0.012) was demonstrated. In conclusion, high OPN expression seems to be related to poor prognosis and MAPK pathway activation, given the association with EGFR and HER2, members of the MAPK signaling pathway.(AU)

A osteopontina é uma glicofosfoproteina implicada em diferentes processos fisiológicos e patológicos, sendo conhecida por estar envolvida na progressão e metástase de vários cânceres nos humanos, no entanto, essa relação é ainda pouco explorada na veterinária. O objetivo deste trabalho foi avaliar a expressão da osteopontina nos carcinomas mamários caninos e sua relação com biomarcadores bem estabelecidos para esta neoplasia. Para isto, foi avaliada a expressão de OPN, EGRH, HER2 e c-Kit juntamente com a taxa de Ki67 em 43 carcinomas mamários. A osteopontina foi expressa pelas células epiteliais neoplásicas em todos os carcinomas, assim como, nas células estromais do microambiente tumoral. Foi demonstrada uma relação entre uma alta expressão de osteopontina e positividade para EGFR (P<0.001) e superexpressão de HER2 (P=0.012). Em conclusão, alta expressão de OPN parece estar relacionada com mau prognóstico e ativação da via MAPK, devido a sua associação com EGRF e HER2, os quais são membros desta via de sinalização.(AU)

Osteopontin expression and its relationship with prognostic biomarkers in canine mammary carcinomas / Expressão de osteopontina e sua relação com biomarcadores prognóstico nos carcinomas mamários caninos

Osteopontin is a glycophosphoprotein implicated in different physiologic and pathologic processes and is known to be involved in progression and metastasis of various cancers in humans, but this relation is still little explored in the veterinary. The aim was to evaluate the expression of osteopontin in canine mammary carcinomas and its relation with well-established canine mammary tumor biomarkers. For that, expression of OPN, EGFR, HER2, and c-Kit were evaluated along with Ki67 rate in 43 mammary carcinomas. Osteopontin was demonstrated to be expressed by neoplastic epithelial cells in all carcinomas as well as in stromal cells from the tumor microenvironment. Relation between high osteopontin expression and EGFR positivity (P<0.001) and HER2 overexpression (P=0.012) was demonstrated. In conclusion, high OPN expression seems to be related to poor prognosis and MAPK pathway activation, given the association with EGFR and HER2, members of the MAPK signaling pathway.(AU)

A osteopontina é uma glicofosfoproteina implicada em diferentes processos fisiológicos e patológicos, sendo conhecida por estar envolvida na progressão e metástase de vários cânceres nos humanos, no entanto, essa relação é ainda pouco explorada na veterinária. O objetivo deste trabalho foi avaliar a expressão da osteopontina nos carcinomas mamários caninos e sua relação com biomarcadores bem estabelecidos para esta neoplasia. Para isto, foi avaliada a expressão de OPN, EGRH, HER2 e c-Kit juntamente com a taxa de Ki67 em 43 carcinomas mamários. A osteopontina foi expressa pelas células epiteliais neoplásicas em todos os carcinomas, assim como, nas células estromais do microambiente tumoral. Foi demonstrada uma relação entre uma alta expressão de osteopontina e positividade para EGFR (P<0.001) e superexpressão de HER2 (P=0.012). Em conclusão, alta expressão de OPN parece estar relacionada com mau prognóstico e ativação da via MAPK, devido a sua associação com EGRF e HER2, os quais são membros desta via de sinalização.(AU)

Clinicopathological and immunohistochemical features of pleomorphic lobular mammary carcinoma in a female dog

This report describes a pleomorphic lobular carcinoma (PLC) case, in an eight-year-old, female Shih Tzu dog, which attended the Veterinary Hospital of the University of Franca, Brazil. A single irregular, non-ulcerated, nonhyperaemic soft nodule, 2 cm in diameter, was observed during the clinical examination. The nodule was adhered within the deep tissues in the right caudal thoracic mammary gland. After removal of the nodule, the histopathologic examination showed that the neoplastic epithelial cells were scattered throughout the stroma in linear patterns, had abundant eosinophiliccytoplasm, eccentric nuclei and cytoplasmatic vacuoles, features compatible with the diagnosis of pleomorphic lobularcarcinoma. An immunohistochemical evaluation was performed in order to better characterise the tumour. Based on thenegative immunoreactivity for hormone receptors and human epidermal growth factor receptor-2 and positive expression for one of the basal markers, the basal-like triple-negative phenotype was characterized. The treatment indicated was chemotherapy with carboplatin; however, the disease progressed, and the patient had an overall survival of 47 days after surgery, confirming the aggressiveness of the disease.

Clinicopathological and immunohistochemical features of pleomorphic lobular mammary carcinoma in a female dog

This report describes a pleomorphic lobular carcinoma (PLC) case, in an eight-year-old, female Shih Tzu dog, which attended the Veterinary Hospital of the University of Franca, Brazil. A single irregular, non-ulcerated, nonhyperaemic soft nodule, 2 cm in diameter, was observed during the clinical examination. The nodule was adhered within the deep tissues in the right caudal thoracic mammary gland. After removal of the nodule, the histopathologic examination showed that the neoplastic epithelial cells were scattered throughout the stroma in linear patterns, had abundant eosinophiliccytoplasm, eccentric nuclei and cytoplasmatic vacuoles, features compatible with the diagnosis of pleomorphic lobularcarcinoma. An immunohistochemical evaluation was performed in order to better characterise the tumour. Based on thenegative immunoreactivity for hormone receptors and human epidermal growth factor receptor-2 and positive expression for one of the basal markers, the basal-like triple-negative phenotype was characterized. The treatment indicated was chemotherapy with carboplatin; however, the disease progressed, and the patient had an overall survival of 47 days after surgery, confirming the aggressiveness of the disease.(AU)

Milk fat globule-epidermal growth factor 8 (MFG-E8) attenuates sepsis-induced acute kidney injury by inhibiting NF-kappa B signaling pathway

Purpose:To explore the effect of milk fat globule-epidermal growth factor 8 (MFG-E8) on sepsis-induced acute kidney injury (SAKI).Methods:Male C57BL/6 mice were randomized to control, sham, CLP, CLP+PBS, and CLP+rmMFG-E8 groups. SAKI was induced by cecal ligation and puncture (CLP). Recombinant mouse MFG-E8 (rmMFG-E8) (20 μg/kg) or PBS (vehicle) was administered intraperitoneally. Blood, urine and renal tissue were collected at 24 h after CLP. Blood samples were tested for serum kidney injury biomarker and cytokines. Urine samples were collected to detect KIM-1, and NGAL. Real-time PCR was tested for Bax and Bcl-2. TUNEL staining was used to determine renal apoptosis. Western blot was used to detect the expression of Bax, Bcl-2, and proteins in the NF-κB pathway.Results:MFG-E8 alleviated SAKI by decreasing serum Cre, BUN, urine KIM-1 and NGAL and by mitigating renal pathological changes significant (p < 0.05). IL-1β, IL-6, TNF-α were significantly inhibited by MFG-E8 (p < 0.05). Apoptosis induced by SAKI was markedly suppressed by MFG-E8. Finally, MFG-E8 attenuated the activation of the NF-𝜅B signaling pathway in SAKI.Conclusion:MFG-E8 has beneficial effects on SAKI, which may be achieved by inhibiting the NF-κB pathway.(AU)

Production of recombinant human epidermal growth factor in Pichia pastoris

This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27 µg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.(AU)

Blocking the PI3K pathway or the presence of high concentrations of EGF inhibits the spontaneous activation of ovine primordial follicles in vitro

The aims of this study were to verify the effects of Epidermal Growth Factor (EGF) on the morphology, primordial follicle activation, growth and proliferation of granulosa cells of ovine follicles cultured in situ, as well as the effect of a PI3K inhibitor on the follicular activation. Ten ovine ovaries were divided into fragments, being one fixed for histological analysis (fresh control). The remaining fragments were cultured for 7 days in control medium (α-MEM+) alone or supplemented with EGF (1, 10, 50, 100 or 200 ng/mL). Follicles were classified as normal or atretic, as primordial or growing, and the oocyte and follicle diameters were measured. PCNA immunohistochemistry was performed in the fresh control and in treatment that showed the bestresults for follicular activation. Pharmacologic inhibition of PI3K activity was performed through pretreatment in media added with 50 μMLY294002 for 1 h. The percentage of normal follicles decreased (P 0.05). In conclusion, PI3K pathway mediates the in vitrospontaneous activation of sheep primordial follicles. Moreover, EGF may act indirectly on follicular activation by promoting granulosa cell proliferation at 1 ng/mL, and EGF inhibited follicle activation in concentrations similar or higher than 10 ng/mL.

Blocking the PI3K pathway or the presence of high concentrations of EGF inhibits the spontaneous activation of ovine primordial follicles in vitro

The aims of this study were to verify the effects of Epidermal Growth Factor (EGF) on the morphology, primordial follicle activation, growth and proliferation of granulosa cells of ovine follicles cultured in situ, as well as the effect of a PI3K inhibitor on the follicular activation. Ten ovine ovaries were divided into fragments, being one fixed for histological analysis (fresh control). The remaining fragments were cultured for 7 days in control medium (α-MEM+) alone or supplemented with EGF (1, 10, 50, 100 or 200 ng/mL). Follicles were classified as normal or atretic, as primordial or growing, and the oocyte and follicle diameters were measured. PCNA immunohistochemistry was performed in the fresh control and in treatment that showed the bestresults for follicular activation. Pharmacologic inhibition of PI3K activity was performed through pretreatment in media added with 50 μMLY294002 for 1 h. The percentage of normal follicles decreased (P < 0.05) after 7 days of culture in all treatments compared to the fresh control. A significantreduction in the percentage of primordial follicles and an increase (P < 0.05) in the growing ones were observed in all treatments compared to fresh control. Furthermore, both the control medium and 1 ng/mL EGF promoted an increase (P < 0.05) in follicular activation compared to other EGF treatments. The PCNA-positive cells in the EGF treatment were higher (P < 0.05) than in fresh control and α-MEM+. Pretreatment of ovarian tissue with PI3K inhibitor significantly inhibited (P < 0.05) α-MEM+-stimulated primordial follicle activation, but had no effect on EGF-stimulated activation (P > 0.05). In conclusion, PI3K pathway mediates the in vitrospontaneous activation of sheep primordial follicles. Moreover, EGF may act indirectly on follicular activation by promoting granulosa cell proliferation at 1 ng/mL, and EGF inhibited follicle activation in concentrations similar or higher than 10 ng/mL.(AU)