Characterisation and evaluation of the regenerative capacity of Stro-4+ enriched bone marrow mesenchymal stromal cells using bovine extracellular matrix hydrogel and a novel biocompatible melt electro-written medical-grade polycaprolactone scaffold.

Many skeletal tissue regenerative strategies centre around the multifunctional properties of bone marrow derived stromal cells (BMSC) or mesenchymal stem/stromal cells (MSC)/bone marrow derived skeletal stem cells (SSC). Specific identification of these particular stem cells has been inconclusive. However, enriching these heterogeneous bone marrow cell populations with characterised skeletal progenitor markers has been a contributing factor in successful skeletal bone regeneration and repair strategies. In the current studies we have isolated, characterised and enriched ovine bone marrow mesenchymal stromal cells (oBMSCs) using a specific antibody, Stro-4, examined their multipotential differentiation capacity and, in translational studies combined Stro-4+ oBMSCs with a bovine extracellular matrix (bECM) hydrogel and a biocompatible melt electro-written medical-grade polycaprolactone scaffold, and tested their bone regenerative capacity in a small in vivo, highly vascularised, chick chorioallantoic membrane (CAM) model and a preclinical, critical-sized ovine segmental tibial defect model. Proliferation rates and CFU-F formation were similar between unselected and Stro-4+ oBMSCs. Col1A1, Col2A1, mSOX-9, PPARG gene expression were upregulated in respective osteogenic, chondrogenic and adipogenic culture conditions compared to basal conditions with no significant difference between Stro-4+ and unselected oBMSCs. In contrast, proteoglycan expression, alkaline phosphatase activity and adipogenesis were significantly upregulated in the Stro-4+ cells. Furthermore, with extended cultures, the oBMSCs had a predisposition to maintain a strong chondrogenic phenotype. In the CAM model Stro-4+ oBMSCs/bECM hydrogel was able to induce bone formation at a femur fracture site compared to bECM hydrogel and control blank defect alone. Translational studies in a critical-sized ovine tibial defect showed autograft samples contained significantly more bone, (4250.63 mm3, SD = 1485.57) than blank (1045.29 mm3, SD = 219.68) ECM-hydrogel (1152.58 mm3, SD = 191.95) and Stro-4+/ECM-hydrogel (1127.95 mm3, SD = 166.44) groups. Stro-4+ oBMSCs demonstrated a potential to aid bone repair in vitro and in a small in vivo bone defect model using select scaffolds. However, critically, translation to a large related preclinical model demonstrated the complexities of bringing small scale reported stem-cell material therapies to a clinically relevant model and thus facilitate progression to the clinic.

The first serological report for genotype C bovine parainfluenza 3 virus in ruminant species of mid-northen Turkey: Traces from the past.

Bovine parainfluenza 3 virus (BPI3V)is one of the most important respiratory pathogens and a leading cause of serious respiratory illnesses in cattle, both independent of and in connection with other pathogens involved in the bovine respiratory disease complex (BRDC). In this study, we aimed to identify the historical circulation of genotype C bovine BPI3V (BPI3Vc) in Turkey using the archival serum samples of domestic ruminants that had been collected from six provinces of northern Anatolia in Turkey between 2009-2010. A total of 896 sera from cattle (n=442), sheep (n=330), and goats (n=124) were randomly selected and screened with a virus neutralization test in order to detect antibodies for BPI3Vc. The overall seropositivity rate was 21.09%, with seropositivity rates for cattle, sheep, and goats of 21.04%, 20.00%, and 24.19%, respectively. Neutralizing antibody titers for selected samples ranged between 1/4 to 1/512. This study represents the first serological study conducted using the first BPI3V isolate of Turkey.

The first serological report for genotype C bovine parainfluenza 3 virus in ruminant species of mid-northen Turkey: Traces from the past

@#Bovine parainfluenza 3 virus (BPI3V)is one of the most important respiratory pathogens and a leading cause of serious respiratory illnesses in cattle, both independent of and in connection with other pathogens involved in the bovine respiratory disease complex (BRDC). In this study, we aimed to identify the historical circulation of genotype C bovine BPI3V (BPI3Vc) in Turkey using the archival serum samples of domestic ruminants that had been collected from six provinces of northern Anatolia in Turkey between 2009-2010. A total of 896 sera from cattle (n=442), sheep (n=330), and goats (n=124) were randomly selected and screened with a virus neutralization test in order to detect antibodies for BPI3Vc. The overall seropositivity rate was 21.09%, with seropositivity rates for cattle, sheep, and goats of 21.04%, 20.00%, and 24.19%, respectively. Neutralizing antibody titers for selected samples ranged between 1/4 to 1/512. This study represents the first serological study conducted using the first BPI3V isolate of Turkey.

Long-term exposure to a continuous 900 MHz electromagnetic field disrupts cerebellar morphology in young adult male rats.

The pathological effects of exposure to an electromagnetic field (EMF) during childhood and adolescence may be greater than those from exposure during adulthood. We investigated possible pathological changes in the cerebellum of adolescent rats exposed to 900 MHz EMF daily for 25 days. We used three groups of six 21-day-old male rats as follows: unexposed control group (Non-EG), sham-exposed group (Sham-EG) and an EMF-exposed group (EMF-EG). EMF-EG rats were exposed to EMF in an EMF cage for 1 h daily from postnatal days 21 through 46. Sham-EG rats were placed in the EMF cage for 1 h daily, but were not subjected to EMF. No procedures were performed on the Non-EG rats. The cerebellums of all animals were removed on postnatal day 47, sectioned and stained with cresyl violet for histopathological and stereological analyses. We found significantly fewer Purkinje cells in the EMF-EG group than in the Non-EG and Sham-EG groups. Histopathological evaluation revealed alteration of normal Purkinje cell arrangement and pathological changes including intense staining of neuron cytoplasm in the EMF-EG group. We found that exposure to continuous 900 MHz EMF for 1 h/day during adolescence can disrupt cerebellar morphology and reduce the number of Purkinje cells in adolescent rats.

Periosteum tissue engineering in an orthotopic in vivo platform.

The periosteum plays a critical role in bone homeostasis and regeneration. It contains a vascular component that provides vital blood supply to the cortical bone and an osteogenic niche that acts as a source of bone-forming cells. Periosteal grafts have shown promise in the regeneration of critical size defects, however their limited availability restricts their widespread clinical application. Only a small number of tissue-engineered periosteum constructs (TEPCs) have been reported in the literature. A current challenge in the development of appropriate TEPCs is a lack of pre-clinical models in which they can reliably be evaluated. In this study, we present a novel periosteum tissue engineering concept utilizing a multiphasic scaffold design in combination with different human cell types for periosteal regeneration in an orthotopic in vivo platform. Human endothelial and bone marrow mesenchymal stem cells (BM-MSCs) were used to mirror both the vascular and osteogenic niche respectively. Immunohistochemistry showed that the BM-MSCs maintained their undifferentiated phenotype. The human endothelial cells developed into mature vessels and connected to host vasculature. The addition of an in vitro engineered endothelial network increased vascularization in comparison to cell-free constructs. Altogether, the results showed that the human TEPC (hTEPC) successfully recapitulated the osteogenic and vascular niche of native periosteum, and that the presented orthotopic xenograft model provides a suitable in vivo environment for evaluating scaffold-based tissue engineering concepts exploiting human cells.

Elastography to assess the effect of varicoceles on testes: a prospective controlled study.

Varicoceles are the most common and treatable cause of male infertility. The pathophysiology of varicoceles primarily includes elevated temperature, adrenal hormone reflux, gonadotoxic metabolite reflux, altered testicular blood flow, antisperm antibody formation and oxidative stress. The diagnosis of a varicocele is mainly clinical. However, a Doppler ultrasound is used to obtain clinical data and to more accurately measure testicular size. Acoustic radiation force impulse (ARFI) is an additional technique to simultaneously show different areas with different densities in a colour-coded image and a B-mode or greyscale image. This can be used for structural analysis of testicular tissue and has become an additional method for detecting pathologic tissue alterations. We enrolled 30 patients who had clinically diagnosed with left varicoceles and male infertility (Group 1). All patients were evaluated by history taking, physical examination, a spermiogram and an endocrine profile. Thirty control patients (Group 2) were randomly chosen from patients who had applied to an andrology clinic for infertility; their physical examinations and laboratory results showed normal findings. Mean elastography results were significantly different between the groups, and significantly lower in patients who had varicoceles. The relationship between hormonal profiles and elastography parameters was calculated as statistically significant negative correlations between FSH and elasticity. Additionally, a negative correlation was determined between varicocele grade and elasticity of testes. In conclusion, our prospective study showed that ARFI imaging may be more useful than palpation for determining early damage of testicular structure by varicoceles.

Effects of quercetin on kidney injury induced by doxorubicin.

OBJECTIVES: The anthracycline antitumor drug doxorubicine causes severe nephrotoxicity in a variety of experimental animals and may be nephrotoxic to humans. The aim of present study was to determine the protective effects of quercetin against doxorubicin-induced kidney injury with light microscopy. METHODS: Forty male Wistar albino rats were divided into four groups: control, doxorubicin, doxorubicin+quercetin and quercetin. A single dose of 20 mg/kg/ i.p. doxorubicin was used to induce injury. Quercetin was administrated orally against doxorubicin toxicity. The kidneys were examined under light microscopy after H-E (hematoxylin-eosin) staining and the changes were scored. RESULTS: Significant tissue injury was observed in doxorubicin-administered group. Among these injuries, renal tubular dilatation, tubular vacuolar changes, glomerular vacuolization, decrease in bowman space, bowman capsule thickening, and interstitial infiltration were evident. However, the injury induced by doxorubicin was attenuated with quercetin administration. DISCUSSION: Quercetin decreased doxorubicin-induced kidney damage (Tab. 1, Fig. 4, Ref. 27).

Drug-drug interaction checking assisted by clinical decision support: a return on investment analysis.

BACKGROUND: Drug-drug interactions (DDIs) are very prevalent in hospitalized patients. OBJECTIVES: To determine the number of DDI alerts, time saved, and time invested after suppressing clinically irrelevant alerts and adding clinical-decision support to relevant alerts. MATERIALS AND METHODS: The most frequently occurring DDIs were evaluated for clinical relevance by a multidisciplinary expert panel. Pharmacist evaluation of relevant DDIs was facilitated using computerized decision support systems (CDSS). During Phase 1, only CDSS-assisted DDI checking was implemented. During Phase 2, CDSS-assisted DDI checking remained in place, and clinically irrelevant DDIs were suppressed. In each phase, the number of alerts and duration of pharmacist DDI checking were compared to conventional DDI checking. In addition, the time invested to implement and configure the CDSS was compared to the time saved using CDSS-assisted DDI checking. RESULTS: CDSS-assisted DDI checking resulted in a daily decrease of DDI checking alerts from 65 to 47 alerts in Phase 1 (P = .03) and from 73 to 33 alerts in Phase 2 (P = .003). DDI checking duration decreased from 15 to 11 minutes (P = .044) and from 15½ to 8½ minutes (P = .001) in Phases 1 and 2, respectively. Almost 298 of the 392 hours required for implementation were invested by pharmacists. An annual timesaving of 30 hours yielded a return on investment of 9.8 years. CONCLUSION: CDSS-assisted DDI checking resulted in a 55% reduction of the number of alerts and a 45% reduction in time spent on DDI checking, yielding a return on investment of almost 10 years. Our approach can be used to refine other drug safety checking modules, increasing the efficiency of checking for drug safety without the need to add more staff pharmacists.

A tattoo pigmented node and breast cancer.

Over the last decade, the axillary SLNB has replaced routine ALND for clinical staging in early breast cancer. Studies describe a potential pitfall in the identification of a true sentinel node during surgery due to lymph node pigmentation secondary to migration of tattoo dye. These pigmented "pseudo-sentinel" nodes, if located superficially in the axilla, may mimic the blue sentinel node on visual inspection, therefore missing the true sentinel node and potentially understaging the patient. Here, we present a case report of a breast cancer patient with a tattoo and discuss the importance of tattoo pigment in the LN (Fig. 1, Ref. 8).

The estimation of the volume of sheep mandibular defects using cone-beam computed tomography images and a stereological method.

OBJECTIVE: The Cavalieri principle of stereological methods is widely used to estimate the volume of structures. Recently in clinical practice, it has become common to use this approach for daily routine purposes. The Cavalieri principle provides quantitative and unbiased volume estimates which are independent of the observer. In the present study, the efficacy of using the Cavalieri principle to estimate the volume of sheep mandibular defects on cone beam CT (CBCT) scans was tested. METHODS: 24 differently sized defects were created on 4 sheep mandibles. Before the defects were created, the outer boundaries of the defects were determined using plaster casts. CBCT scans of the defects were taken. The scans were reconstructed in the coronal plane and sections of 0.2 mm thickness with 0.2 mm and 0.4 mm intervals were obtained. The volume of each defect was estimated using the Cavalieri principle. The models were created using light-body silicone for the estimation of the actual volume of each defect. They were immersed in water using a pycnometer and the actual volumes were obtained on the basis of the Archimedean principle. The actual and estimated volumes of the defects were compared using the Wilcoxon signed-rank test. RESULTS: The results showed that the volumes from the Cavalieri estimates did not differ from the actual volumes of the defects (P > 0.05). CONCLUSION: We concluded that the volume of mandibular defects can be accurately estimated using the Cavalieri principle on images from a CBCT scan.

Chronic prenatal exposure to the 900 megahertz electromagnetic field induces pyramidal cell loss in the hippocampus of newborn rats.

Widespread use of mobile phones which are a major source of electromagnetic fields might affect living organisms. However, there has been no investigation concerning prenatal exposure to electromagnetic fields or their roles in the development of the pyramidal cells of the cornu ammonis in postnatal life. Two groups of pregnant rats, a control group and an experimental group, that were exposed to an electromagnetic field were used. For obtaining electromagnetic field offspring, the pregnant rats were exposed to 900 megahertz electromagnetic fields during the 1-19th gestation days. There were no actions performed on the control group during the same period. The offspring rats were spontaneously delivered--control group (n = 6) and electromagnetic field group (n = 6). Offspring were sacrificed for stereological analyses at the end of the 4th week. Pyramidal cell number in rat cornu ammonis was estimated using the optical fractionator technique. It was found that 900 megahertz of electromagnetic field significantly reduced the total pyramidal cell number in the cornu ammonis of the electromagnetic field group (P < 0.001). Therefore, although its exact mechanism is not clear, it is suggested that pyramidal cell loss in the cornu ammonis could be due to the 900 megahertz electromagnetic field exposure in the prenatal period.

Achieving efficient anti-adhesion in re-mediastinoscopy: an experimental study in rats.

OBJECTIVE: Re-mediastinoscopy can be a difficult procedure due to fibrosis in the mediastinum. We have investigated the effect of an anti-adhesive barrier agent "hylan B gel" on the formation of adhesions after dissection in the superior mediastinum in a rat model. METHODS: The study was conducted in 70 male Sprague-Dawley rats weighing 300 g. The strap muscles were divided by a midline cervical incision. The anterior and lateral aspects of the trachea were dissected to the level of the carina. Hylan B gel (Sepragel Sinus, Genzyme Company, Redgefield, NJ, USA) was used to create an anti-adhesive barrier. The rats who were subjected to surgery were sacrificed on postoperative days 14 and 28. All rats were previously divided into 5 groups: Sham group (n = 10); Group 1 (n = 15) surgery alone, sacrificed on day 14; Group 2 (n = 15) surgery and Sepragel, sacrificed on day 14; Group 3 (n = 15) surgery alone, sacrificed on day 28; Group 4 (n = 15) surgery and Sepragel, sacrificed on day 28. Histopathological analysis was performed to study the effect of the anti-adhesive agent. Scores were calculated based on collagen fibrosis, fibroblasts, granulation tissue, muscle alterations/inflammation, histiocytes, mononuclear giant cells, inflammation and vascular proliferation. RESULTS: Collagen fibrosis and fibroblast scores, which represent adhesions, were significantly higher in Groups 1, 2, 3, 4 than in the Sham group ( P < 0.01). The degree of adhesions on day 14 was found to be higher in Group 1 compared to Group 2 ( P < 0.01). Adhesions on day 28 were higher in Group 3 compared to Group 4 ( P < 0.01). There were no differences between groups with regard to foreign body reactions ( P > 0.05). CONCLUSION: Adhesions causing technical difficulty during re-mediastinoscopy can be reduced by the application of anti-adhesive agents during the primary procedure.

Histopathological effects of bipolar vessel sealing devices on liver parenchyma and comparison with suture method: an experimental study.

BACKGROUND: LigaSureand SurgRx are bipolar vessel sealing devices providing hemostasis by denaturating collagen and elastin from the vessel wall and surrounding connective tissue. We aimed to histopathologically evaluate the lateral injury during rat liver resection with LigaSure and SurgRx. METHODS: Suture technique was used in group A, LigaSure was used in group B and SurgRx was used in group C to resect one lobe of liver from midline. One of the resected pieces was histopathologically examined the same day and the other piece was left in the animal to be examined at the 7th day. Relaparotomy was performed at the 7th day. RESULTS: Necrosis, exudate formation, chronic inflammation, histiocytes and fibroblasts scores were significantly lower in SurgRx group compared to the other groups. CONCLUSION: Our findings suggest that LigaSure and SurgRx can be safely used in liver resection as compared to suture technique while SurgRx was superior than LigaSure in inflammatory response as it causes lower lateral thermal injury and inflammatory scores probably due to its different technical properties.