Effects of hyperprolactinemia and ovariectomy on the tibial epiphyseal growth plate and bone formation in mice

PURPOSE: To evaluate the effects of ovariectomy and the hyperprolactinemia procedure in the tibial epiphyseal growth plate of female mice.METHODS: In this study, the epiphyseal growth plate of ovariectomized (OVX) and/or rendered hyperprolactinemic female mice by 50 days of treatment with 200 μg metoclopramide (M) was evaluated morphologically, morphometrically and immuno-histochemically. Forty female and adult mice were divided into four groups according to treatment: V group - animals treated with saline solution; H group - hyperprolactinemic animals; Ovx/V group - ovariectomized animals and treated with saline solution; Ovx/H group - hyperprolactinemic and ovariectomized animals. After the treatment period, the animals were sacrificed, tibia was removed and fixed in 10% buffered formalin and decalcified in 10% formic acid. The material was immersed in paraffin and subjected to histological processing in paraffin. The sections were stained with Masson's trichrome and immunohistochemistry was carried out for the pro-apoptotic protein BCL-2. The images for the morphological and morphometric study were analyzed with the imaging program AxioVision 4.8 (Carl-Zeiss(r), Germany).RESULTS: The combination of hyperprolactinemia and the ovariectomy procedure decreased the number of resting chondrocytes 1.5-fold, the number of proliferative chondrocytes 1.8-fold; the percentage of resting cartilage 2.4-fold and the percentage of trabecular bone 2.1-fold, compared with respective control animals.CONCLUSION: The procedure of ovariectomy combined with the metoclopramide-induced hyperprolactinemia in female mice has showed marked bone degeneration due to significant decrease of cell proliferation in the epiphyseal growth plate and bone formation.

Are alveolar bone changes a determinant factor for "cara inchada" in cattle?

In order to study possible alterations of the skeleton which might play a role in the pathogenesis of the periodontitis of "cara inchada" in young cattle, ribs from 20 affected calves, 2 to 10 months old, were examined. Electrolytically decalcified longitudinal sections of the costochondral junction and cross sections through the corpus costae, stained with Haematoxylin-Eosin, were studied. In five calves, longitudinal sections of the proximal humerus were examined as well. The status of mineralization was checked by microradiograms. Systemic alteration of the skeleton due to disturbances of mineral metabolism could not be shown in any of the animals. In seven 2 to 4 months old calves, no bone changes were found. The reduced osteogenesis in six 3 to 5 months old calves and the reduced osteogenesis and diminished chondral growth in seven 5 to 10 months old calves are therefore a consequence of the disease. The results show that the development of the alveolar bone was not defective, so this cannot be a determinant factor for the development of the periodontitis of "cara inchada" in cattle.

Bone changes caused by experimental Solanum malacoxylon poisoning in rabbits

The aim of this study was to describe the bone changes observed after a daily oral administration of the calcinogenic plant Solanum malacoxylon (syn. S. glaucophyllum) (Sm) during 9 days. The Sm-poisoned rabbits had an increase of bone resorption in the endosteal surface of the cortical zone and also in the surface covered by osteoblasts of the primary and secondary spongiosa of the trabecular bone compartment. Moreover, the epiphyseal growth plates in long bones appeared narrower than in the control rabbits, with reduction of the proliferative and hyperthrophic chondrocyte zones. The electron microscopic study revealed a significant decrease of proteoglycans in the hyperthrophic chondrocyte zone evidenced by a significant reduction of rutenium red positive granules in the poisoned rabbit. Altogether, these data suggest that cell differentiation may play a pivotal role in the pathogenesis of Sm-induced bone lesions.

Effect of bone morphogenetic protein-7 (BMP-7) on in vitro survival of caprine preantral follicles

This study was conducted in order to verify the effect of different concentrations of BMP-7 in the in vitro survival and development of caprine preantral follicles. Fragments of caprine ovarian cortical tissue were cultured for 1 or 7 days in Minimum Essential Medium (MEM+) supplemented with different concentrations of BMP-7 (1, 10, 50 or 100ng/ml). Non-cultured fragments or those cultured for 1 or 7 days were processed for classical histology and transmission electron microscopy (TEM). Parameters such as follicular survival, activation and growth were evaluated. The results showed that, after 1 or 7 days of culture, the percentage of morphologically normal follicles was significantly reduced in all treatments when compared with fresh control, except at 1ng/ml of BMP-7 for 1 day. In addition, the concentration of 10ng/ml of BMP-7 significantly increases follicular diameter from day 1 to 7 of culture. There was no influence of the other concentrations of BMP-7 regarding to the follicular and oocyte diameter. Ultrastructure studies confirmed follicular integrity after 7 days of culture in 1ng/ml BMP-7. In conclusion, small concentrations of BMP-7 can improve the survival and growth of caprine preantral follicles during in vitro culture.

Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro

This study investigated the effects of bone morphogenetic protein 6 (BMP-6) on in vitro primordial follicle development in goats. Samples of goat ovarian cortex were cultured in vitro for 1 or 7 days in Minimum Essential Medium (control medium) supplemented with different concentrations of BMP-6. Follicle survival, activation and growth were evaluated through histology and transmission electron microscopy (TEM). After 7 days of culture, histological analysis demonstrated that BMP-6 enhanced the percentages of atretic primordial follicles when compared to fresh control (day 0). Nevertheless, BMP-6 increased follicular and oocyte diameter during both culture periods. As the culture period progressed from day 1 to day 7, a significant increase in follicle diameter was observed with 1 or 50ng/ml BMP-6. However, on the contrary to that observed with the control medium TEM revealed that follicles cultured for up to 7 days with 1 or 50ng/ml BMP-6 had evident signs of atresia. In conclusion, this study demonstrated that BMP-6 negatively affects the survival and ultrastructure of goat primordial follicles.

Primary bone neoplasms in dogs: 90 cases

A retrospective study of necropsy and biopsy cases of 90 primary bone tumors (89 malignant and one benign) in dogs received over a period of 22 years at the Laboratório de Patologia Veterinária, Universidade Federal de Santa Maria, was performed. Osteosarcoma was the most prevalent bone tumor, accounting for 86.7% of all malignant primary bone neoplasms diagnosed. Most cases occurred in dogs of large and giant breeds with ages between 6 and 10-years-old. The neoplasms involved mainly the appendicular skeleton, and were 3.5 times more prevalent in the forelimbs than in the hindlimbs. Osteoblastic osteosarcoma was the predominant histological subtype. Epidemiological and pathological findings of osteosarcomas are reported and discussed.

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

The objective of this study was to evaluate the culture of equine bone marrow mononuclear fraction and adipose tissue - derived stromal vascular fraction cells in two different cell culture media. Five adult horses were submitted to bone marrow aspiration from the sternum, and then from the adipose tissue of the gluteal region near the base of the tail. Mononuclear fraction and stromal vascular fraction were isolated from the samples and cultivated in DMEM medium supplemented with 10% fetal bovine serum or in AIM-V medium. The cultures were observed once a week with an inverted microscope, to perform a qualitative analysis of the morphology of the cells as well as the general appearance of the cell culture. Colony-forming units (CFU) were counted on days 5, 15 and 25 of cell culture. During the first week of culture, differences were observed between the samples from the same source maintained in different culture media. The number of colonies was significantly higher in samples of bone marrow in relation to samples of adipose tissue.

Lead poisoning in cattle and chickens in the state of Pará, Brazil

The present study describes the occurrence of lead poisoning in cattle and chickens in Pará, Brazil. In a lot composed of 80 calves from a dairy herd, 10 animals became sick and nine died, but one animal recovered after being removed from the paddock. Upon inspection of this paddock, the presence of truck batteries used to store energy captured by solar panels was found. The clinical signs observed in calves included difficult breathing, nasal discharge, excessive salivation, corneal opacity, pushing of the head against objects and recumbency. The chickens had decreased oviposition and produced eggs with thin or malformed shells. The necropsy findings of the cattle, as well as the histopathological changes observed, were of little significance except for one animal that showed mild astrocytosis histopathology in the cerebral cortex. In one of the chickens, renal histopathology showed mild multifocal acute tubular necrosis. The mean lead concentrations in the livers and kidneys of the cattle were 93.91mg/kg and 209.76mg/kg, respectively, and the mean concentration detected in chicken livers was 105.02mg/kg. It was concluded that the source of lead poisoning in these calves and chickens were the truck battery plates, which were within reach of the animals.

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

Mesenchymal stem cells (MSC) are increasingly being proposed as a therapeutic option for treatment of a variety of different diseases in human and veterinary medicine. Stem cells have been isolated from feline bone marrow, however, very few data exist about the morphology of these cells and no data were found about the morphometry of feline bone marrow-derived MSCs (BM-MSCs). The objectives of this study were the isolation, growth evaluation, differentiation potential and characterization of feline BM-MSCs by their morphological and morphometric characteristics. in vitro differentiation assays were conducted to confirm the multipotency of feline MSC, as assessed by their ability to differentiate into three cell lineages (osteoblasts, chondrocytes, and adipocytes). To evaluate morphological and morphometric characteristics the cells are maintained in culture. Cells were observed with light microscope, with association of dyes, and they were measured at 24, 48, 72 and 120h of culture (P1 and P3). The non-parametric ANOVA test for independent samples was performed and the means were compared by Tukey's test. On average, the number of mononuclear cells obtained was 12.29 (±6.05x10(6)) cells/mL of bone marrow. Morphologically, BM-MSCs were long and fusiforms, and squamous with abundant cytoplasm. In the morphometric study of the cells, it was observed a significant increase in average length of cells during the first passage. The cell lengths were 106.97±38.16µm and 177.91±71.61µm, respectively, at first and third passages (24 h). The cell widths were 30.79±16.75 µm and 40.18±20.46µm, respectively, at first and third passages (24 h).The nucleus length of the feline BM-MSCs at P1 increased from 16.28µm (24h) to 21.29µm (120h). However, at P3, the nucleus length was 26.35µm (24h) and 25.22µm (120h). This information could be important for future application and use of feline BM-MSCs.

Israelin strategisen kommunikaation järjestelyt ja toteutus Cast Lead -operaatiossa

Verkottuneessa ja globaalissa maailmassa ei pelkkään kineettiseen vaikuttamiseen perustuva sodankäynti ole tuottanut enää toivottuja tuloksia ja siksi sekä valtiolliset että ei-valtiolliset toimijat ovatkin kääntäneet katseensa muihin keinoihin. Vaikuttaminen on siirtymässä maa-likeskeisyydestä mielikeskeisyyteen, jossa maaleina ovat kohteen arvot, asenteet, ajatukset ja lopulta identiteetti. Näihin vaikuttamalla halutaan muuttaa kohteen käyttäytymistä omalta kannalta suotuisaan suuntaan. Teknologiset vallankumoukset ovat lisänneet informaation määrää eksponentiaalisesti ja tä-hän globaaliin informaatioon on alati kasvavalla joukolla pääsy lähes milloin ja mistä tahan-sa. Jokainen informaatioon käsiksi pääsevä voi myös ryhtyä tuottamaan informaatiota mui-den saataville, ja tämä viestinnällinen mullistus on vaikuttanut radikaalisti myös länsimaiseen sotataitoon informaation noustessa yhdeksi ratkaisevaksi tekijäksi nykyajan ja tulevaisuuden konflikteissa. Tapahtumista, joista vielä sata vuotta sitten kuultiin vasta päiviä tai viikkoja niiden jälkeen, saadaan tänä päivänä nähdä reaaliaikaista kuvavirtaa mobiilipäätelaitteen vä-lityksellä. Tämä mullistus on pakottanut niin valtiolliset kuin ei-valtiollisetkin toimijat kiin-nittämään entistä enemmän huomiota taisteluun informaatiosta informaatioympäristössä sekä yleisön mielikuvista ja mielipiteistä. Fyysisestä kontaktista voittajana selvinnyt osapuoli voi jäädä häviäjän rooliin, mikäli taistelu informaatiorintamalla hävitään. Totuus on toisarvoista, mielikuvat ratkaisevat. Jotta taistelu mielikuvista, ajatuksista ja asenteista olisi mahdollisimman tehokasta, täytyy kunkin osapuo-len organisoida omat ponnistuksensa resurssien käytön optimoimiseksi karsien päällekkäi-syydet ja ristiriidat omista sanomistaan. Kaikkien eri tahojen täytyy siis sovittaa omat vies-tinsä palvelemaan ylemmän tahon tavoitteita päätyen aina politiikan määrittämään tavoittee- seen. Tätä kutsutaan strategiseksi kommunikaatioksi, jota voidaan pitää ajattelua ohjaavana mallina tai jopa eräänlaisena sotataidollisena paradigmana. Tässä työssä tutkittiin Israelin suorittamaa sotilaallista operaatiota Gazan kaistaleella vuosien 2008 – 2009 vaihteessa. Operaatiosta käytetään yleisesti länsimaissa nimeä Cast Lead. Ope-raation tarkoituksena oli iskeä Gazassa toimivaa, länsimaiden terroristijärjestöksi katsomaa Hamas-järjestöä vastaan ja lopettaa sen tekemät raketti- ja kranaatti-iskut Israeliin. Tutki-muksen tarkoituksena oli selvittää, miten strateginen kommunikaatio oli järjestetty ja toteu-tettu Israelissa operaatioon liittyen. Työ on jatkoa tutkijan esiupseerikurssilla 64 laatimalle työlle ”Psykologinen vaikuttaminen CAST LEAD -operaatiossa”. Lähdemateriaalin avulla muodostettiin strategisen kommunikaation viitekehys, jota käytettiin analyysirunkona teoriaohjaavalle sisällön analyysille. Tutkittavaa operaatiota käsittelevästä avoimista lähteistä kerättyä aineistoa analysoitiin strategisen kommunikaation viitekehyksen kautta keräten havaintoja viitekehyksen eri osa-alueiden esiintymisestä. Tällä tavalla kyettiin muodostamaan kuva Israelin strategisen kommunikaation organisoinnista, eri toimijoista se-kä niiden vastuista ja eri tavoista, joilla viestejä välitettiin millekin kohdeyleisölle. Samalla pyrittiin hahmottelemaan Israelin narratiivia tutkittavaan operaatioon. Työ jakautuu kolmeen osaan. Ensimmäisessä osassa luvuissa yksi ja kaksi käsitellään tutki-muksen teoreettiset taustat, menetelmät ja metodit sekä määritellään keskeisimmät käsitteet. Luvussa kolme käsitellään strategisen kommunikaation osatekijät käsiteanalyysin kautta. Toisessa osassa luvuissa neljä ja viisi käsitellään operaation kulku ja tausta pääpiirteittäin se-kä jaotellaan strategisen kommunikaation viitekehyksen avulla tehdyt havainnot kuvaamaan strategisen kommunikaation toteutumista operaatiossa. Kolmannessa osassa luvussa kuusi käsitellään johtopäätökset sekä arvioidaan tutkimuksen toteutumista sekä jatkotutkimustarpeita. Strategisen kommunikaation viitekehys muodostettiin ulkomaisten ja suomalaisten lähdete-osten avulla. Strategisen kommunikaation käsiteanalyysissä käsiteltiin strategiaa, kommuni-kaatiota sekä näiden taustalla olevia teorioita eri lähteiden avulla ja siten pyrittiin muodosta-maan laaja-alainen kuva siitä, mitä strateginen kommunikaatio on ja miten se toimii. Tutki-muksen aineisto koottiin avoimista lähteistä internetin eri mediatalojen sivuilta ja sitä täy-dennettiin virallisten ja epävirallisten tahojen sivustojen tarjoamalla informaatiolla. Aineiston perusteella kyettiin muodostamaan kuva Israelin strategisen kommunikaation or- ganisoinnista vastuineen sekä hahmottelemaan viestejä, teemoja ja narratiivia. Strategisen kommunikaation vaikuttavuutta tässä operaatiossa on mahdotonta mitata aukottomasti, koska tapaustutkimukselle luonteenomaisesti käsiteltävänä oli ainutlaatuinen tosielämän tapahtuma ja siten suoraa vertailukohtaa ei ole. Verrokkina voidaan kuitenkin käyttää aiempia Israelin sotilaallisia toimia, joissa informaatioympäristöön ei oltu kiinnitetty yhtä paljon huomiota. Näin ollen voidaan vetää johtopäätöksiä operaation vaikuttavuudesta vertailemalla esimer-kiksi näiden ajallisesti läheisten operaatioiden aiheuttamia reaktioita maailmalla.

Development of porous glass-fiber reinforced composite for bone implants. Evaluation of antimicrobial effect and implant fixation

Cranial bone reconstructions are necessary for correcting large skull bone defects due to trauma, tumors, infections and craniotomies. Traditional synthetic implant materials include solid or mesh titanium, various plastics and ceramics. Recently, biostable glass-fiber reinforced composites (FRC), which are based on bifunctional methacrylate resin, were introduced as novel implant solution. FRCs were originally developed and clinically used in dental applications. As a result of further in vitro and in vivo testing, these composites were also approved for clinical use in cranial surgery. To date, reconstructions of large bone defects were performed in 35 patients. This thesis is dedicated to the development of a novel FRC-based implant for cranial reconstructions. The proposed multi-component implant consists of three main parts: (i) porous FRC structure; (ii) bioactive glass granules embedded between FRC layers and (iii) a silver-polysaccharide nanocomposite coating. The porosity of the FRC structure should allow bone ingrowth. Bioactive glass as an osteopromotive material is expected to stimulate the formation of new bone. The polysaccharide coating is expected to prevent bacterial colonization of the implant. The FRC implants developed in this study are based on the porous network of randomly-oriented E-glass fibers bound together by non-resorbable photopolymerizable methacrylate resin. These structures had a total porosity of 10–70 volume %, of which > 70% were open pores. The pore sizes > 100 μm were in the biologically-relevant range (50-400 μm), which is essential for vascularization and bone ingrowth. Bone ingrowth into these structures was simulated by imbedding of porous FRC specimens in gypsum. Results of push-out tests indicated the increase in the shear strength and fracture toughness of the interface with the increase in the total porosity of FRC specimens. The osteopromotive effect of bioactive glass is based on its dissolution in the physiological environment. Here, calcium and phosphate ions, released from the glass, precipitated on the glass surface and its proximity (the FRC) and formed bone-like apatite. The biomineralization of the FRC structure, due to the bioactive glass reactions, was studied in Simulated Body Fluid (SBF) in static and dynamic conditions. An antimicrobial, non-cytotoxic polysaccharide coating, containing silver nanoparticles, was obtained through strong electrostatic interactions with the surface of FRC. In in vitro conditions the lactose-modified chitosan (chitlac) coating showed no signs of degradation within seven days of exposure to lysozyme or one day to hydrogen peroxide (H2O2). The antimicrobial efficacy of the coating was tested against Staphylococcus aureus and Pseudomonas aeruginosa. The contact-active coating had an excellent short time antimicrobial effect. The coating neither affected the initial adhesion of microorganisms to the implant surface nor the biofilm formation after 24 h and 72 h of incubation. Silver ions released to the aqueous environment led to a reduction of bacterial growth in the culture medium.

Protocol for collection and separation of bone marrow mononuclear cells in Chlorocebus aethiops

Abstract: Chlorocebus aethiops is a species of non-human primate frequently used in biomedical research. Some research involves this species as an experimental model for various diseases and possible treatment with stem cells. The bone marrow is one of the main sources of these cells and provides easy access. The aim of this study was to standardize the protocol of collection and separation of bone marrow in C. aethiops. Ten animals were submitted to puncture of bone marrow with access to the iliac crest and cell separation by density gradient. The bone marrow of C. aethiops had an average of 97% viability. From the results achieved, we can conclude that C. aethiops is an excellent model to obtain and isolate mononuclear cells from bone marrow, fostering several studies in the field of cell therapy.

Effectiveness of plant growth promoting rhizobacteria in facilitating lead and nutrient uptake by little seed canary grass

We investigated the effectiveness of Nitroxin inoculation on lead (Pb) and nutrient uptakes by little seed canary grass. The factors tested included inoculation (or not) with Nitroxin and different soil concentrations of Pb (0, 200, 400 and 800mgPbkg-1 soil). Increasing soil concentrations of Pb decreased stem, leaf and root dry weights. Shoot phosphorus concentrations increased in parallel with increasing soil Pb concentrations. Nitroxin inoculation did not alter the phosphorus concentration of the roots. The Pb translocation factor was >1 in inoculated treatments in the Pb soil concentration range of 200 to 400mgkg-1; the translocation factor for 800mgPbkg‑1 with no inoculation of Nitroxin was, however, <1. Our results indicated that the Pb bioaccumulation factor for little seed canary grass was <1, indicating that it is a Pb excluding plant.

Genotoxic activity of the insecticide Nuvacron (Monocrotophos) detected by the micronucleus test in bone marrow erythrocytes of mice and in CHO cells

The organophosphorus insecticide Nuvacron (Monocrotophos) is a very toxic agent widely utilized in Brazilian agriculture. To evaluate the clastogenic potential of this insecticide, in vivo and in vitro micronucleus (MN) assay experiments were carried out on Swiss mice and on Chinese hamster ovary (CHO) cells, respectively. Nuvacron administered at doses of 2.5 and 5.0 mg/kg induced a statistically significant increase in the frequencies of MN detected in polychromatic bone marrow erythrocytes from animals (six/group) treated ip 24 h before. Exponentially growing CHAO cells were treated continuously (16h) with Nuvacron diluted in water to final concentrations of 1, 10, 100, 200, and 400 mug/ml. Three experiments were carried out using the cytokinesis-block method and a total of 6000 binucleated cells were scored to determine MN frequencies. A statistically significant increase in the frequencies of MN was observed for the cells treated with 1 and 10 mug/ ml Nuvacron. A marked decrease in cell proliferation rates was observed for CHO cultures treated with higher concentrations. These data demonstrate that Nuvacron has a genotoxic effect on both in vivo and in vitro mammalian test systems.

Lead (Pb2+) and cadmium (Cd2+) inhibit the dipsogenic action of central beta-adrenergic stimulation by isoproterenol

We have previously demonstrated that acute third ventricle injections of both Pb2+ and Cd2+ impair the dipsogenic response elicited by three different situations: dehydration and central cholinergic or angiotensinergic stimulation. ß-Adrenergic activation is part of the multifactorial integrated systems operating in drinking behavior control in the central nervous system. In the present study acute third ventricle injections of Pb2+ (3, 30 and 300 pmol/rat) or Cd2+ (0.3, 3 and 30 pmol/rat) blocked the dipsogenic response induced by third ventricle injections of isoproterenol (ISO; 160 nmol/rat) in a dose-dependent manner. Normohydrated animals receiving ISO + NaAc (sodium acetate) or saline (controls) displayed a high water intake after 120 min (ISO + saline = 5.78 ± 0.54 ml/100 g; ISO + NaAc = 6.00 ± 0.6 ml/100 g). After the same period, animals receiving ISO but pretreated with PbAc at the highest dose employed (300 pmol/rat) drank 0.78 ± 0.23 ml/100 g while those receiving ISO and pretreated with the highest dose of CdCl2 (30 pmol/rat) presented a water intake of 0.7 ± 0.30 ml/100 g. Third ventricle injections of CdCl2 (3 nmol/rat) or PbAc (3 nmol/rat) did not modify food intake in rats deprived of food for 24 h. Thus, general central nervous system depression explaining the antidipsogenic action of the metals can be safely excluded. It is concluded that both Pb2+ and Cd2+ inhibit water intake induced by central ß-adrenergic stimulation