Siyami Karahan

In-vivo efficacy of toltrazuril on experimentally induced Toxoplasma gondii tissue cysts in lambs: A novel strategy for prevention of human exposure to meat-borne toxoplasmosis

The aim of the present study was to investigate in vivo efficacy of toltrazuril on Toxoplasma gondii tissue cysts following induction of chronic toxoplasmosis in 4-week-old lambs (n=27) by inoculation of 1×10(5) T. gondii ME 49 strain oocysts (day 0). Beginning at the 15th day after inoculation, lambs in Group T20 and Group T40 were given toltrazuril orally 2 times, once every week (Baycox 5%, Bayer Animal Health) at a dose of 20 mg/kg and 40 mg/kg, respectively. Positive control (PC) lambs were not given any therapy, and 2 clinically healthy non-infected lambs were used as negative controls (Group NC). Two out of 9 lambs in PC group (oocyst inoculated but non-treated) were killed on toltrazuril treatment days (day 15 and 22) to evaluate the tissue cyst presence in their muscles. On day 90, the remaining 25 lambs were necropsied, and samples from the brain and 11 different muscle groups were collected. The tissues were examined for the presence of tissue cysts by histopathology, immunohistochemistry, nested-PCR and percoll gradient centrifugation. Anti-T. gondii antibodies were screened by IFAT throughout the experiment. The increased T. gondii seropositivity beginning from the 15th day of inoculation remained steady at Day 45 and Day 90 in Groups PC while it was significantly lower at Day 90 in toltrazuril receiving groups. In toltrazuril treated groups, histopathological findings included degenerative changes in the cyst wall, complete macrophage invasion to the cysts, and reduction or removal of the cysts in toto. Four out of 9 lambs (44.4%) in both toltrazuril treated group (Group T20 and T40) did not contain tissue cyst in any examined tissues while all positive control animals had T. gondii tissue cysts at least in one muscle group. The toltrazuril treatment efficacy on the cyst presence was determined as 44.4%. The number of the cysts in the musculature was significantly different between non-treated and toltrazuril treated lambs (X(2)=6.613; p=0.037). For the total number of cysts, the positive control lambs had higher number of cysts compared to both toltrazuril treated lambs (T20 and T40) (X(2)=5.629; p=0.018 and X(2)=5.629; p=0.018, respectively) while there were no differences between Group T20 and Group T40 (X(2)=0.000; p=1.000). According to PCR results, the brain and M. semitendinosus were positive in all 7 control lambs while 12 out of 18 lambs were positive in toltrazuril treated lambs. In conclusion, the results are promising as the toltrazuril treated lambs had markedly less parasite counts compared to those of untreated lambs. Further research should be conducted to reveal if toltrazuril treatment in sheep could be used as a strategy to minimize the cyst exposure of humans through consumption of raw or undercooked mutton.

Polyethyleneimine brushes effectively inhibit encrustation on polyurethane ureteral stents both in dynamic bioreactor and in vivo.

Polyurethane (PU) ureteral stents have been widely used as biomedical devices to aid the flow of the urine. Due to the biofilm formation and encrustation complications it has been hindered their long term clinical usage. To overcome these complications, in this study, cationic polyethyleneimine (PEI) brushes grafted on PU stents and their performances were tested both in a dynamic biofilm reactor system (in vitro) and in a rat model (in vivo). Thus, we hypothesized that PEI brushes inhibit bacterial adhesion owing to the dynamic motion of brushes in liquid environment. In addition, cationic structure of PEI disrupts the membrane and so kills the bacteria on time of contact. Cationic PEI brushes decreased the biofilm formation up to 2 orders of magnitude and approximately 50% of encrustation amount in respect to unmodified PU, in vitro. In addition, according to Atomic Absorption Spectroscopy (AAS) results, approximately 90% of encrustation was inhibited on in vivo animal models. Decrease in encrustation was clearly observed on the stents obtained from rat model, by Scanning Electron Microscopy (SEM). Also, histological evaluations showed that; PEI brush grafting decreased host tissue inflammation in close relation to decrease in biofilm formation and encrustation. As a results; dual effect of anti-adhesive and contact-killing antibacterial strategy showed high efficiency on PEI brushes grafted PU stents both in vitro and in vivo.

Fabrication and characterization of gold-nanoparticles/chitosan film: a scaffold for L929-fibroblasts

Abstract The objective of the present study was to fabricate a gold nanoparticle crosslinked chitosan (Ch/AuNPs) composite film simple and to evaluate its use as a carrier matrix for L929- fibroblasts. L929- fibroblasts were seeded either onto Ch or Ch/AuNPs scaffolds. The Ch/AuNPs scaffold exhibited a higher cell proliferation and growth rate. The cytotoxicity test determined trypan blue staining indicated that Ch scaffolds devoid of AuNPs expressed almost no toxicity while the Ch/AuNPs composite scaffolds expressed a very limited toxicity only at higher doses. The Ch/AuNPs scaffold promotes cell attachment, growth and proliferation with almost no cytotoxicity.

The fine structures of Cystocaulus ocreatus (Nematoda: Protostrongylidae) and the related lung pathology

SummaryIn the present study, the fine surface structures of Cystocaulus ocreatus and its worm as well as brood nodules obtained from the lung of Akkaraman sheep are demonstrated by scanning electron and light microscopy. The mouth of the parasite obtained from the worm nodules is located at the anterior tip and encircled by a collar of tissue. The mouth opening appears as three slits conjoined in the shape of a capital case letter Y, creating three lips. The mouth is surrounded by six papillae just behind the collar. The anterior end was similar in both sexes and covered by numerous wrinkles. Female parasite has a bell-shaped pro-vagina. The posterior end of the female parasite was sharply pointed. The male parasite has a bursa. In the worm nodule, curled parasites were enclosed by a capsula and no eggs and larvae of the parasite were observed. In the brood nodule, heavily affected alveoli and bronchioles contained excessive amounts of larvae and eggs.

Characterization and Evaluation of Triamcinolone, Raloxifene, and Their Dual-Loaded Microspheres as Prospective Local Treatment System in Rheumatic Rat Joints

In this study, injectable microspheres were developed for the local treatment of joint degeneration in rheumatoid arthritis (RA). Microspheres loaded with triamcinolone (TA), a corticosteroid drug, and/or raloxifene (Ral), a cartilage regenerative drug, were prepared with a biodegradable and biocompatible polymer, polycaprolactone (PCL). Microspheres were optimized for particle size, structural properties, drug release, and loading properties. In vitro release of Ral was very slow because of the low solubility of the drug and hydrophobic nature of PCL. However, when coloaded with TA, both drugs were released at higher amounts compared with their single forms. Smallest particle sizes were obtained in dual drug-loaded microspheres. In vitro cytotoxicity tests showed biocompatibility of microspheres. In vivo bioefficacy of these microspheres was also examined in adjuvant-induced arthritis model in rats. In vivo histological studies of control groups showed development of RA with high median lesion score (5.0). Compared with control and intra-articular free drug injections, microsphere treatment groups showed lower lesion scores and better healing outcomes in histological evaluations. Results suggest that a controlled delivery system of TA and RAL by a single injection in inflamed joints holds promise for healing and suppressing inflammation.

Immunohistochemical localisation of 3β-hydroxysteroid dehydrogenase in Sarcocystis spp.

Abstract3β-Hydroxysteroid-dehydrogenase (3β-HSD) is an isoenzyme that catalyses an essential step in the synthesis of all classes of active steroid hormones. The presence of steroid hormones of the vertebrate type in invertebrates is acknowledged in addition to a group of steroid-like hormones called ecdysteroids that were present in arthropods and helminths. In the present study, 3β-HSD was detected in the bradyzoites enclosed in sarcocysts of Sarcocystis spp. with immunohistochemistry. The results suggest that self-originating steroid hormones may play important roles in the development of Sarcocystis spp., and possibly in the regulation of the reciprocal immune interaction between the host and these parasites.

Characterization of chondrocytes cultured on catechin-loaded alginate-chitosan scaffolds

Abstract Bovine chondrocytes were seeded into scaffolds of a high molecular weight chitosan and alginate with a pore size of 50–350 μm with or without catechin. In polymerase chain reaction (PCR), unlike type II, collagen type I was no longer expressed at day 14. The DNA content increased until day 8 and began declining, indicating cell detachment. The GAG content increased during the first 12 days. The percentage of round and collagen type II immunoreactive cells increased over the time. Catechin has some protective properties on chondrocytes seeded on the alginate-chitosan scaffolds during the first 12 days by means of DNA and chondrocyte morphology (p < 0.05).

Collagen Based Multilayer Scaffolds for Meniscus Tissue Engineering: In Vivo Test Results

Meniscus is an important component of the knee joint since it performs several crucial functions such as shock absorption, load bearing and transmission, maintenance of joint stability, and lubrication. The results of common meniscal injury repair approaches are not fully satisfactory with low mechanical properties and long regeneration times. A 3D collagen-based construct consisting of multilayers of lyophilized sponges separated by electrospun fibrous mats was prepared previously to serve as a substitute for meniscus. Mechanical properties of the construct were studied in vitro and a 3 to 4 fold increase was observed when a double crosslinking method was used. Rabbit meniscal cells were cultured in vitro, expanded and seeded onto the polymer scaffolds. 2 weeks later the substitute was implanted to the medial compartment of the rabbit knee joint. The implants were studied 3 and 10 weeks after transplantation. Histological and microscopical characterization showed a significant difference between the groups (Group I: control; Group II: cell free substitute and Group III: cell seeded substitute) with Week3 sample scores. Group III healing score was significantly lower than I and II, which was probably due to the the fibrous tissue surrounding the cell seeded material but this resulted in lower immunological responses. Moreover, the scores decreased from Week3 to Week10 indicating healing. Even though there were no statistically significant differences, the lowest values were observed with the tissue engineered substitute. Therefore, it can be concluded that in vivo studies showed the potential of the cell seeded artificial meniscus.