In vitro assessment of motility and proliferation of human osteogenic cells on different isolated extracellular matrix components compared with enamel matrix derivative by continuous single-cell observation

OBJECTIVES: The composition of the extracellular matrix (ECM) plays a substantial role in bone remodelling, fracture healing and osseointegration of dental implants by regulating proliferation, migration and finally differentiation of osteogenic cell populations. Emdogain, a composition of an enamel matrix derivative (EMD), has been introduced as a potential candidate to promote tissue regeneration. We investigated whether EMD could serve as a potential promoter of cell proliferation and motility as a dynamic cell response and compared the results with the ubiquitous single ECM components type I collagen and laminin. MATERIAL AND METHODS: In the investigation presented, we used a continuous observation method for the analysis of migratory and proliferative patterns of individual cells. We analyzed the response of four osteoblastic cell lines to specific extracellular ligands (type I collagen, laminin and EMD) over a period of 24 h compared with untreated glass surface and bovine serum albumin (BSA) as control groups. RESULTS: Type I collagen and laminin promoted cell motility significantly compared with the control groups and, in part, compared with EMD as well. The analysis of all 451 investigated cells revealed the following mean values for cell motiliy: untreated glass (n=99): 5.46+/-2.74 microm/h, BSA (n=89): 6.35+/-2.43 microm/h, type I collagen (n=108): 8.77+/-3.42 microm/h, laminin (n=74): 9.89+/-5.10 microm/h and EMD (n=81): 7.92+/-3.35 microm/h. Proliferation rates on the different surfaces were heterogenous for all investigated cell lines and varied from 0% to 50% within 24 h without a correlation to cell motility. CONCLUSION: In our study, EMD promotes cell motility better than the control groups. The two investigated single ECM components type I collagen and laminin promoted cell motility superior to EMD. This supports the hypothesis that EMD promotes a less mobile but more differentiated osteogenic phenotype.     

Epstein-Barr virus BMRF-2 and BDLF-3 expression in hairy leukoplakia

Hairy leukoplakia (HL) is a lesion found on the side of the tongue of immunocompromised individuals, including those with human immunodeficiency virus (HIV) infection. The lesion has unique histopathologic features and is characterised by high-level Epstein-Barr virus (EBV) replication, multiple EBV strains, and extensive inter-and intra-strain recombination. Expression of EBV genes spanning the entire viral life cycle from latency-associated genes to late, replicative genes has been detected in the lesion. HL thus provides a unique opportunity to study EBV expression in oral epithelium, and to study expression of novel EBV genes. We therefore constructed a cDNA library from an HL biopsy and detected expression of two genes not previously described in vivo: BMRF-2 and BDLF-3. Sequence analysis of the cDNAs revealed few amino acid changes from the B95-8 sequence. Expression of both genes was localized to the lower prickle cell layer of the tongue epithelium. BMRF-2 protein expression was primarily detected in the cell nuclei of the upper prickle cell layer. BDLF-3 protein expression was observed in the perinuclear space and Golgi compartment. The function of these proteins is currently under investigation.     

Prevalence of Enterococcus faecalis at multiple oral sites in endodontic patients using culture and PCR

There are conflicting data on the occurrence of Enterococcus faecalis in the oral cavity of endodontic patients. This study investigated the prevalence of E. faecalis in multi-site oral samples (n = 136) from 41 endodontic patients using culture and polymerase chain reaction (PCR). Additionally, culturable strains were investigated for virulence traits. Overall, E. faecalis was detected in at least one tongue, oral rinse, or gingival sulcus sample in 68% of patients and in the root canals only in 5% of patients. In 21 patients from whom samples were obtained from all four sites, E. faecalis was detected in more tongue than gingival sulcus, oral rinse, and root canal samples (43, 14, 10, and 10%, respectively; p = 0.0148, chi(2)), and in proportionally greater numbers of patients with gingivitis/periodontitis compared to healthy periodontium (73% versus 20%; p = 0.03, Fisher's exact test). PCR was more sensitive than culture in detecting E. faecalis in oral samples (32% and 4%, respectively; p < 0.0001, McNemar's test). Multiple virulence traits were identified in culturable strains.     

The inability of Streptococcus mutans and Lactobacillus acidophilus to form a biofilm in vitro on dentine pretreated with ozone

Background: The use of ozone therapy in the treatment of dental caries is equivocal. The aim of this study was to use an in vitro model to determine the effects of prior ozone application to dentine on biofilm formation and to measure any associated reduction in bacteria viability. Methods: Twenty dentine discs were bonded to the bases of 5 mL polycarbonate screw top vials. Ten dentine discs were infused with ozone for 40 seconds, 10 samples remained untreated as a control. The vials were filled with nutrient medium, sterilized and placed into the outflow from a continuous chemostat culture of Streptococcus mutans and Lactobacillus acidophilus for four weeks. At the conclusion of the experiment bacterial growth was monitored by taking optical density readings of the growth medium in each vial and the outer surface of the dentine specimens were examined by scanning electron microscopy as shown by SEM analysis. Results: Ozone infusion prevented biofilm formation on all the treated samples while there was substantial biofilm present on the control specimens. While the average optical density of the control specimens was almost twice that of the ozone infused dentine (0.710 for the control with a SD of 0.288 and 0.446 for the ozonated samples with a SD of 0.371), the results were not significant (p > 0.05). Conclusions: This preliminary study has shown that the infusion of ozone into non‐carious dentine prevented biofilm formation in vitro from S. mutans and L. acidophilus over a four‐week period. The possibility exists that ozone treatment may alter the surface wettability of dentine through reaction with organic constituents.     

Effects of Attention and Perceptual Uncertainty on Cerebellar Activity During Visual Motion Perception

Recent clinical and neuroimaging studies have revealed that the human cerebellum plays a role in visual motion perception, but the nature of its contribution to this function is not understood. Some reports suggest that the cerebellum might facilitate motion perception by aiding attentive tracking of visual objects. Others have identified a particular role for the cerebellum in discriminating motion signals in perceptually uncertain conditions. Here, we used functional magnetic resonance imaging to determine the degree to which cerebellar involvement in visual motion perception can be explained by a role in sustained attentive tracking of moving stimuli in contrast to a role in visual motion discrimination. While holding the visual displays constant, we manipulated attention by having participants attend covertly to a field of random-dot motion or a colored spot at fixation. Perceptual uncertainty was manipulated by varying the percentage of signal dots contained within the random-dot arrays. We found that attention to motion under high perceptual uncertainty was associated with strong activity in left cerebellar lobules VI and VII. By contrast, attending to motion under low perceptual uncertainty did not cause differential activation in the cerebellum. We found no evidence to support the suggestion that the cerebellum is involved in simple attentive tracking of salient moving objects. Instead, our results indicate that specific subregions of the cerebellum are involved in facilitating the detection and discrimination of task-relevant moving objects under conditions of high perceptual uncertainty. We conclude that the cerebellum aids motion perception under conditions of high perceptual demand.     

Criteria for successful correction of thoracolumbar/lumbar curves in AIS patients: results of risk model calculations using target outcomes and failure analysis

Introduction

Failure to select the appropriate lowest instrumented vertebra (LIV) in selective lumbar fusion (SLF) for thoracolumbar/lumbar curves (LC) can result in adding-on in the lumbar curve (LC) or the need for fusion extension due to a decompensating thoracic curve (TC). The selection criteria that predict optimal outcomes still need to be refined. The objectives of the current study were to identify risk factors for failure of anterior scoliosis correction and fusion (ASF) as well as predictors of optimal outcomes and ASF efficacy for SLF.

Materials and methods

A retrospective review of all patients (n = 245) with AIS who had anterior SLF at one institution was conducted. Optimal outcomes were defined as a target LC ≤20° and a target TC ≤30°. The distance from the LIV to the SV was recorded. An increase in the LIV adjacent level disc angulation (LIVDA) ≥5° was defined as adding-on. An increase in the TC at follow-up was defined as TC-progression. Stepwise univariate and multivariate linear and logistic regression analyses were performed to identify criteria predicting the target LC and TC. A total of 68 % of the patients had the LIV at SV-2 (=2 levels above stable vertebra).

Results

The patients’ average age was 17 years, the average fusion length was 4.6 levels, and the average follow-up time was 32 months. The preoperative LC was 49 ± 14°, the LC-bending was 22 ± 13° (57 ± 18 %), and the follow-up LC was 25 ± 10°. LC correction was 59 ± 17 % (p < 0.01). The preoperative TC was 39 ± 13°, the TC-bending was 21 ± 12°, and the follow-up TC was 29 ± 13°. The TC-correction was 32 ± 19 % (p < 0.01). At follow-up, 85 patients (35 %) had an LC ≤20°, 110 patients (45 %) had a TC ≤30°. The follow-up LC and an LC ≤20° were predicted by LC-bending (p < 0.01, r = 0.6), preoperative LC (p < 0.01, r = 0.6). The logistic regression models could define patients at risk for failing the target LC ≤20° or TC ≤30°. At follow-up, TC ≤30° was best predicted by the preoperative TC (p < 0.01, r = 0.8; OR 1.2) and TC-bending (p < 0.01, r = 0.8; OR 1.06), with the logistic regression model revealing a correct prediction in 84 % of all cases. Among the patients, 8 % required late posterior surgery. Patients achieving the target LC ≤20° had a significantly reduced risk for failure (p = 0.01). Selecting an LIV at SV-1 vs. SV-2 significantly increased the chance of achieving a target LC ≤20° (p = 0.01) and reduced the risk of adding-on (p < 0.01). Predictors for failure also included a high preoperative LC (p = 0.02; OR 0.97), TC-bending (p < 0.01), and preoperative TC (p = 0.01). A cut-off in the failure risk analysis was established at a TC of 38°. Additionally, a significant cut-off for risk of adding-on was established at LIVDA <3.5°.

Conclusion

A high chance of achieving a target LC ≤20° and a low risk of revision was dependent on LC-bending, preoperative LC and TC, and a LIV at SV-1 with non-parallel LIVDA. Our risk model analysis may support the selection of a safe LIV to achieve the target LC.     

Long-term survival of participants in the prostate ：ancer prevention trial

The Prostate Cancer Prevention Trial （PCPT） is a seminal study in the field of urology. More than 10years after its initial publication, updated data from this trial continue to shape our understanding of prostate cancer. Among the major findings from the PCPT has been the demonstration that prostate cancer is common in men with prostate-specific antigen （PSA） once thought to be in the normal range,~ finasteride prevents the development of benign prostatic hypertrophy,2 it increases the sensitivity of PSA3 and digital rectal examination.4 Furthermore the PCPT helped to establish the link between erectile dysfunction and cardiovascular disease,5 and perhaps most importantly finasteride demonstrated a 25% relative risk reduction in the diagnosis of prostate cancer compared with placebo.     

Radium-223 in metastatic castration resistant prostate cancer

In 2004, docetaxel was approved for the treatment of metastatic castration-resistant prostate cancer （mCRPC）. For the next several years, there was a lull in drug approvals. However, from 2010 onwards, 5 additional therapies have been approved on the basis of showing a survival benefit in phase III studies. These agents include sipuleuceI-T, cabazitaxel, abiraterone, enzalutamide and （most recently） radium-223. Amongst radiopharmaceuticals currently used for advanced prostate cancer （e.g. samarium-153 and strontium-89）, radium-223 possesses several unique properties. As an alpha-emitting compound, the agent produces a high-energy output over a short range, facilitating selective destruction of tissue within the bone in the region of osteoblastic lesions while sparing surrounding normal tissue. The current review will outline biological rationale for radium-223 and also provide an overview of preclinical and clinical development of the agent. Rational sequencing of radium-223 and combinations, in the increasingly complex landscape of mCRPC will be discussed, along with factors influencing clinical implementation.     

Antiphospholipid syndrome and perioperative hemostatic management of cardiac valvular surgery

Hemostatic aspects of antiphospholipid syndrome (APS) present unique challenges to clinicians and laboratory personnel alike, particularly in the perioperative period. These challenges are especially evident in patients requiring cardiac valve replacement surgery. However, the literature outlining the optimal approach in such patients is limited. We present the case of a 25-year-old woman with severe aortic regurgitation as a result of APS with particular reference to the precautions necessary during perioperative care. Particularly important are the prevention of thrombotic or hemorrhagic complications, management of associated thrombocytopenia, and laboratory methods of perioperative anticoagulation monitoring in the setting of prolonged clotting times.     

In vivo 3D MRI of insect brain: cerebral development during metamorphosis of Manduca sexta

High-resolution 3D MRI of male pupae of Manduca sexta was performed at 2.35 T in order to evaluate its potential for an in vivo characterization of insect brain during metamorphosis. T1-weighted 3D FLASH (TR/TE = 20/7.8 ms, 25 degrees flip angle) and T2-weighted 3D fast SE MRI data sets (TR/TEeff = 3000/100 ms) were acquired at different developmental stages with an isotropic resolution of 100 microm. Both T1- and T2-weighted 3D MRI allowed for the identification of cerebral structures such as the antennal nerve, antennal and optical lobe, and central brain. Pronounced developmental alterations of the morphology were observed during metamorphosis. The results demonstrate the feasibility of 3D MRI at nanoliter resolution to identify major brain systems of M. sexta and respective changes during pupal development from caterpillar to sphinx moth. Together with the use of suitable contrast agents, this approach may provide new ways for studying the axonal connectivity and neural function of the developing insect brain.