Search

Mitochondrial DNA has been investigated for nearly fifty years, but many aspects of the maintenance of this essential small genome remain unknown. Like any genome, mammalian mitochondrial DNA requires the function of topoisomerases to counter and regulate the topological tension arising during replication, transcription, segregation, and repair. However, the functions of the different mitochondrial topoisomerases are poorly understood. Here, we investigate the role of Topoisomerase 3α (Top3α) in mtDNA replication and transcription, providing evidence that this enzyme, previously reported to act in mtDNA segregation, also participates in mtDNA replication fork progression. Top3α knockdown caused replication fork stalling, increased mtDNA catenation and decreased mtDNA levels. Overexpression in contrast induced abundant double-strand breaks around the replication origin OH and abortion of early replication, while at the same time improving the resolution of mtDNA replication termination intermediates. Both Top3α knockdown and overexpression affected mitochondrial RNA transcription, leading to a decrease in steady-state levels of mitochondrial transcripts. Together, our results indicate that the mitochondrial isoform of Top3α is not only involved in mtDNA segregation, as reported previously, but also supports the progression of the replication fork. Mitochondrial Top3α is also influencing the progression of transcription, with its absence affecting downstream transcript levels.

Background Two invasive group A streptococcus (iGAS) infection outbreaks occurred in Montreal in 2016 and 2017; one in a long-term care facility (typeemm118) and one in the community, primarily involving homeless people (typeemm74). Objective To describe two recent iGAS outbreaks in Montreal and highlight the challenges in dealing with these outbreaks and the need to tailor the public health response to control them. Methodology All cases of iGAS were investigated and the isolates were sent to the laboratory foremmtyping. In both outbreaks, cases of superficial group Astreptococcus(GAS) infection were identified, through 1) systematic case detection accompanied by screening for asymptomatic carriers among residents and employees of the long-term care facility and 2) sentinel surveillance among homeless people. Visits were made to community organizations providing homeless services (including shelters) and social networks were analyzed to establish whether there were any links among cases of GAS infection (both invasive and noninvasive) and locations frequented. In both outbreaks, recommendations were made to service providers regarding enhancement of infection prevention and control measures. Results In the long-term care facility, five cases of typeemm118 iGAS were identified over a 22-month period, one of which resulted in death. All residents were screened and no carriers were identified. Among the employees, 81 (65%) were screened and fourcarriers were identified. Of those, one was a carrier of typeemm118 GAS. All carriers were treated, and subsequent follow-up sampling on three carriers (including the one withemm118) was negative.In the community, 23 cases of typeemm74 iGAS were detected over a 16-month period, four of which resulted in death. Half of the cases (n=12) were described as homeless, and six others were users of services for the homeless. Sentinel surveillance of superficial infections yielded 64 cultures with GAS, chiefly on the skin, including 51 (80%) of typeemm74. An analysis of the social networks revealed the large number and variety of resources for the homeless used by the cases. Visits to the community organizations providing homeless services revealed the heterogeneity and precariousness of some of these services, the difficulties encountered in applying adequate health and hygiene measures, and the high degree of mobility amongst those who use these services. Conclusion The detection and control of iGAS outbreaks in both long-term care establishments and among community organizations providing homeless services are very complex. An outbreak of iGAS can develop in the background over a long time and be easily overlooked despite cases being admitted to the hospital.Emmtyping and systematic research of previous cases of iGAS are essential tools for the detection and characterization of outbreaks. Close cooperation among public health agencies, clinical teams, community organizations and laboratories is essential for proper monitoring and the reduction of GAS transmission in the community and health care settings.

BACKGROUNDS: Hospitalizations associated with hepatitis C virus (HCV) infection and liver disease increased on average by 6.0% per year from 2004 to 2010 in Canada and were projected (in 2010) to increase by another 4% by 2016. The first generation of direct-acting antivirals (DAAs) became available in 2012. In 2014, a second generation of effective and well-tolerated DAA therapy was authorized in Canada. The impact of DAA therapy on the HCV-associated disease burden in Canada has not been documented. OBJECTIVES: To assess the potential impact of DAA therapy on the disease burden by a) comparing the actual hospitalization rates associated with HCV infection and liver disease following the introduction of DAAs in Canada with the 2010 baseline projection and b) documenting the associated uptake of anti-HCV therapy. METHODS: The hospital records of inpatients diagnosed with chronic HCV and chronic liver disease were extracted from the Canadian Discharge Abstract Database (DAD) by fiscal year for 2004–2016. We compared the actual number of hospitalizations to the baseline projection by year and for selected 5-year birth cohorts (1925–1989). The monthly number of new prescriptions for anti-HCV regimens was extracted from the IQVIA CDH CompuScript database (formerly IMS Health), aggregated to annual levels by age group and compared with hospitalization trends. RESULTS: Compared to the baseline projection, there was a slight reduction in hospitalizations in 2014/15 and 2015/16. This slight reduction was followed by a more significant decline in 2016/17 (32% below expected; 95% confidence interval [CI]: 27%–37%). The largest declines were observed for patients born before 1960 (age 55 or older) at 40% below expected in 2016/17. The number of new anti-HCV prescriptions increased from 5,484 in fiscal year 2012/13 to a peak of 17,775 in 2015/2016. The number of new prescriptions corresponds to approximately 1.3 and five times the number of hospitalizations in 2012/13 and 2015/16, respectively. CONCLUSIONS: In Canada there has been a modest decrease in HCV and liver-related hospitalizations following a significant increase in uptake of second-generation DAAs in 2015. However, the burden is still high. Linked health administrative databases created to monitor the disease burden in the new treatment era should provide additional insight with the linkage of treatment history and disease stage to individual outcomes.

Background Gonorrhea is the second most commonly reported sexually transmitted infection (STI) in Canada after chlamydia, and rates for this STI have been increasing since 1997. Objective To summarize trends observed in gonorrhea rates for 2010-2015 in Canada. Methods Laboratory-confirmed cases of gonorrhea are reported to the Public Health Agency of Canada (PHAC) by all of the Canadian provinces and territories. The overall national rate was computed, as were rates per sex, age group and province/territory. Results In 2015, 19,845 cases of gonorrhea were reported in Canada, corresponding to a rate of 55.4 cases per 100,000 population and a 65.4% increase from 2010 (33.5 cases per 100,000 population). Males had consistently higher rates than did females (70.2 per 100,000 versus 40.6 per 100,000 in 2015) and faster rising rates (85.2% versus 39.5% in 2010-2015). Rates among adults 60 years and older increased faster than rates among younger people, although the highest rates were among those 15-29 years of age. The Northwest Territories, Nunavut and Yukon had the highest gonorrhea rates in 2015. Conclusion Males, adolescents and young adults continue to represent the majority of gonorrhea cases. Research is needed to better understand the current trends in gonorrhea infection in order to maintain, evaluate and improve primary and secondary STI prevention activities.

Background Chlamydia is the most commonly reported notifiable sexually transmitted infection in Canada. Rates have been steadily increasing since 1997. Objective To summarize trends in chlamydia rates for the period 2010-2015 in Canada. Methods Laboratory-confirmed cases of chlamydia were reported to the Public Health Agency of Canada by all the Canadian provinces and territories. The overall national rate was computed, as were rates per sex, age group and province/territory. Results In 2015, a total of 116,499 cases of chlamydia were reported in Canada, corresponding to a rate of 325 cases per 100,000 population. Females accounted for the majority (two-thirds) of chlamydia infections from 2010 to 2015. However, rates among males rose faster during this time period. Youth and young adults aged 15-29 years had the highest rates in 2015. While increased rates were observed over time for most age groups, adults aged 40 years and older had the greatest increase (51%) between 2010 and 2015. Chlamydia rates increased in most provinces during this period, with the highest rates being reported by the Northwest Territories and Nunavut in 2015. Conclusion Between 2010 and 2015, chlamydia rates increased by 16.7% and were highest among females and young adults. Although a number of factors may account for this rising trend, the possibility of a true increase in incidence cannot be ruled out. Ongoing monitoring of chlamydia and research into the reasons for the observed changes will help guide sexually transmitted infection (STI) prevention and control activities.

Background Syphilis is the third most commonly reported notifiable sexually transmitted infection (STI) in Canada, following chlamydia and gonorrhea, respectively. Rates of this STI have been rising rapidly in Canada since 2001. Objective To summarize trends observed in syphilis rates for 2010 to 2015 in Canada. Methods Laboratory-confirmed cases of infectious syphilis and early congenital syphilis were reported to the Public Health Agency of Canada by all of the Canadian provinces and territories. National infectious syphilis rates were computed, as were rates per sex, age group and province/territory. Rates of congenital syphilis were also calculated. Results From 2010 to 2015, the rate of infectious syphilis in Canada increased by 85.6%, from 5.0 to 9.3 cases per 100,000 population. In 2015, a total of 3,321 cases of infectious syphilis were reported, mainly in males (93.7%), among whom the rate was 17.5 cases per 100,000 males versus 1.2 per 100,000 females. The rate also rose faster among males in 2010-2015, a 90.2% increase versus 27.8% among females. Individuals aged 20-39 years had the highest rates. Across the provinces and territories, the highest rates of infectious syphilis were in Nunavut, British Columbia and Manitoba.​: The rate of congenital syphilis decreased from 2010 to 2014 (1.6 to 0.3 cases per 100,000 live births) before increasing to 1.5 cases per 100,000 live births in 2015, which corresponds to six reported cases. Conclusion Rates of syphilis continue to rise in Canada, especially among young men, and this is consistent with trends in the United States of America and European Union. Based on data from Canada and from these regions, the sexual behaviour of men who have sex with men (MSM) is thought to be a major risk factor for syphilis.

Background Two invasive group A streptococcus (iGAS) infection outbreaks occurred in Montreal in 2016 and 2017; one in a long-term care facility (typeemm118) and one in the community, primarily involving homeless people (typeemm74). Objective To describe two recent iGAS outbreaks in Montréal and highlight the challenges in dealing with these outbreaks and the need to tailor the public health response to control them. Methodology All cases of iGAS were investigated and the isolates were sent to the laboratory foremmtyping. In both outbreaks, cases of superficial group Astreptococcus(GAS) infection were identified, through 1) systematic case detection accompanied by screening for asymptomatic carriers among residents and employees of the long-term care facility and 2) sentinel surveillance among homeless people. Visits were made to community organizations providing homeless services (including shelters) and social networks were analyzed to establish whether there were any links among cases of GAS infection (both invasive and noninvasive) and locations frequented. In both outbreaks, recommendations were made to service providers regarding enhancement of infection prevention and control measures. Results In the long-term care facility, five cases of typeemm118 iGAS were identified over a 22-month period, one of which resulted in death. All residents were screened and no carriers were identified. Among the employees, 81 (65%) were screened and fourcarriers were identified. Of those, one was a carrier of typeemm118 GAS. All carriers were treated, and subsequent follow-up sampling on three carriers (including the one withemm118) was negative. In the community, 23 cases of typeemm74 iGAS were detected over a 16-month period, four of which resulted in death. Half of the cases (n=12) were described as homeless, and six others were users of services for the homeless. Sentinel surveillance of superficial infections yielded 64 cultures with GAS, chiefly on the skin, including 51 (80%) of typeemm74. An analysis of the social networks revealed the large number and variety of resources for the homeless used by the cases. Visits to the community organizations providing homeless services revealed the heterogeneity and precariousness of some of these services, the difficulties encountered in applying adequate health and hygiene measures, and the high degree of mobility amongst those who use these services. Conclusion The detection and control of iGAS outbreaks in both long-term care establishments and among community organizations providing homeless services are very complex. An outbreak of iGAS can develop in the background over a long time and be easily overlooked despite cases being admitted to the hospital.Emmtyping and systematic research of previous cases of iGAS are essential tools for the detection and characterization of outbreaks. Close cooperation among public health agencies, clinical teams, community organizations and laboratories is essential for proper monitoring and the reduction of GAS transmission in the community and health care settings.

In this paper, we provide a novel approach that uses a generative adversarial network to produce synthetic network traffic. The intent is to leverage this synthetic data to improve the robustness of machine learning algorithms that perform analysis on communication networks. In our experimental results, we demonstrate that a generative adversarial network can construct samples of network traffic that are statistically similar to an original set of reference samples. Additionally, we provide insight into the performance of our approach when evaluating different varieties of generative adversarial networks for their ability to produce and converge to realistic output.

Background Unintentional perioperative hypothermia causes serious adverse effects to surgical patients. Thermal suit (T-Balance®) is an option for passive warming perioperatively. We hypothesized that the thermal suit will not maintain normothermia more efficiently than conventional cotton clothes when also other preventive procedures against unintentional hypothermia are used. Methods One hundred patients were recruited to this prospective, randomized trial. They were allocated to the Thermal Suit group or a Control group wearing conventional hospital cotton clothes. All patients received our institution's standard treatment against unintentional hypothermia including a warming mattress, a forced-air upper body warming blanket and a warming device for intravenous fluids. Eardrum temperature was measured pre-operatively. In the operating room and post-anaesthesia care unit temperatures were measured from four locations: oesophagus, left axilla, dorsal surface of the left middle finger and dorsum of the left foot. The primary outcome measure was temperature change during robotic-assisted laparoscopic radical prostatectomy. Results The temperatures of 96 patients were analysed. There was no difference in mean core temperatures, axillary temperatures or skin temperatures on the finger between the groups. Only foot dorsum temperatures were significantly lower in the Thermal Suit group. Intraoperative temperature changes were similar in both groups. In the post-anaesthesia care unit temperature changes were minimal and they did not differ between the groups. Conclusion Provided that standard preventive procedures in maintaining normothermia are effective the thermal suit does not provide any additional benefit over conventional cotton clothes during robotic-assisted laparoscopic radical prostatectomy.

Background — Randomized trials evaluating efficacy of local infiltration analgesia (LIA) have been published but many of these lack standardized analgesics. There is a paucity of reports on the effects of LIA on functional capability and quality of life. Methods — 56 patients undergoing unilateral total knee arthroplasty (TKA) were randomized into 2 groups in this placebo-controlled study with 12-month follow-up. In the LIA group, a mixture of levobupivacaine (150 mg), ketorolac (30 mg), and adrenaline (0.5 mg) was infiltrated periarticularly. In the placebo group, infiltration contained saline. 4 different patient-reported outcome measures (PROMs) were used for evaluation of functional outcome and quality of life. Results — During the first 48 hours postoperatively, patients in the LIA group used less oxycodone than patients in the placebo group in both cumulative and time-interval follow-up. The effect was most significant during the first 6 postoperative hours. The PROMs were similar between the groups during the 1-year follow-up. Interpretation — Single periarticular infiltration reduced the amount of oxycodone used and enabled adequate pain management in conjunction with standardized peroral medication without adverse effects. No clinically marked effects on the functional outcome after TKA were detected.

Summary The possibilities of human pluripotent stem cell-derived neural cells from the basic research tool to a treatment option in regenerative medicine have been well recognized. These cells also offer an interesting tool for in vitro models of neuronal networks to be used for drug screening and neurotoxicological studies and for patient/disease specific in vitro models. Here, as aiming to develop a reductionistic in vitro human neuronal network model, we tested whether human embryonic stem cell (hESC)-derived neural cells could be cultured in human cerebrospinal fluid (CSF) in order to better mimic the in vivo conditions. Our results showed that CSF altered the differentiation of hESC-derived neural cells towards glial cells at the expense of neuronal differentiation. The proliferation rate was reduced in CSF cultures. However, even though the use of CSF as the culture medium altered the glial vs. neuronal differentiation rate, the pre-existing spontaneous activity of the neuronal networks persisted throughout the study. These results suggest that it is possible to develop fully human cell and culture-based environments that can further be modified for various in vitro modeling purposes.

Background Electroencephalogram (EEG)-based depth of anaesthesia monitoring is susceptible to contaminating electromyographic (EMG) activity. Many authorities have suggested that anaesthesiologists using these monitors should interpret the raw EEG waveform seen on the anaesthesia monitor. Methods In 34 patients anaesthetized with propofol using two doses of rocuronium (0.6 and 1.2 mg/kg), we studied whether the EMG arousal can be detected visually on the anaesthesia monitor. The Bispectral Index (BIS) and Entropy biosignals on the monitor were recorded with a video camera, and the one-channel EEG recorded by the Entropy strip was collected on a laptop computer. The recordings and the one-channel EEG were analyzed offline by two experts (anaesthesiologist and neurophysiologist), both with a long experience on anaesthesia-related EEG. Results EMG arousal existed in 14/34 and 13/33 patients in the BIS and Entropy biosignals, respectively. The anaesthesiologist detected EMG on the monitor in 7/14 patients with BIS (sensitivity 50%) and in 4/13 patients with Entropy (31%). The clinical neurophysiologist detected EMG in 6/14 (43%) patients with BIS and in 5/13 (38%) with Entropy. The specificity of the EMG analyses was 55 and 65% with BIS, and 85 and 90% with Entropy. EMG arousal was detected in BIS biosignal in 10/17 and 4/17 patients with 0.6 and 1.2 mg/kg doses of rocuronium (P = 0.04). Conclusions In contrast to many EEG phenomena, EMG activity cannot be accurately detected visually from the raw EEG on the anaesthesia monitor. Further development in the quality of the anaesthesia monitors is warranted.

Background Sugammadex is designed to antagonize neuromuscular blockade (NMB) induced by rocuronium or vecuronium. In clinical practice, we have noticed a rise in the numerical values of bispectral index (BIS) and Entropy, two electroencephalogram (EEG) – based depth of anesthesia monitors, during the reversal of the NMB with sugammadex. The aim of this prospective, randomized, double-blind study was to test this impression and to compare the effects of sugammadex and neostigmine on the BIS and Entropy values during the reversal of the NMB. Methods Thirty patients undergoing gynecological operations were studied. Patients were anesthetized with target-controlled infusions of propofol and remifentanil, and rocuronium was used to induce NMB. After operation, during light propofol-remifentanil anesthesia, NMB was antagonized with sugammadex or neostigmine. During the following 5 min, the numerical values of BIS, BIS electromyographic (BIS EMG) and Entropy were recorded on a laptop computer, as well as the biosignal recorded by the Entropy strip. The Entropy biosignal was studied off-line both in time and frequency domain to see if NMB reversal causes changes in EEG. Results In some patients, administration of sugammadex or neostigmine caused a significant rise in the numerical values of BIS, BIS EMG and Entropy. This phenomenon was most likely caused by increased electromyographic (EMG) activity. The administration of sugammadex or neostigmine appeared to have only minimal effect on EEG. Conclusion The EMG contamination of EEG causes BIS and Entropy values to rise during reversal of rocuronium-induced NMB in light propofol-remifentanil anesthesia.

Purpose of this study was to compare bioactive glass and autogenous bone as a bone substitute material in tibial plateau fractures. We designed a prospective, randomized study consisting of 25 consecutive operatively treated patients with depressed unilateral tibial comminuted plateau fracture (AO classification 41 B2 and B3).14 patients (7 females, 7 males, mean age 57 years, range 25-82) were randomized in the bioglass group (BG) and 11 patients (6 females, 5 males, mean age 50 years, range 31-82) served as autogenous bone control group (AB). Clinical examination of the patients was performed at 3 and 12 months, patients' subjective and functional results were evaluated at 12 months. Radiological analysis was performed preoperatively, immediately postoperatively and at 3 and 12 months. The postoperative redepression for both studied groups was 1 mm until 3 months and remained unchanged at 12 months. No differences were identified in the subjective evaluation, functional tests and clinical examination between the two groups during 1 year follow-up. We conclude that bioactive glass granules can be clinically used as filler material instead of autogenous bone in the lateral tibial plateau compression fractures.

Polymethylmethacrylate (PMMA) has been used in many orthopedic and dental applications since the 1960s. Biocompatibility of newly developed surface porous fiber reinforced (SPFR) PMMA based composite has not been previously proven in cell culture environment. Analysis of rat bone marrow stromal cells grown on the different test materials showed only little difference in normalized cell activity or bone sialoprotein (BSP) production between the test materials, but the osteocalcin (OC) levels remained higher (P < 0.015-0.005) through out the test with SPFR-material when compared to tissue culture poly styrene (TCPS). The cells grown on SP-FRC material also showed highest calcium depletion from the culture medium (P < 0.026-0.001) when compared to all other test substrates. SEM images of the cultured samples confirmed that all the materials enabled cell spreading and growth on their surface, but the roughened surface remarkably enhanced this process of cell attachment, division and calcified nodule formation. This study shows that the SP-FRC composite material does not elicit harmful/toxic reactions in cell cultures more than neutral TCPS and can be considered biocompatible. The material possesses good capabilities to form new mineralized tissue onto its surface, and through that a possibility to bond directly to bone. Rough surface seems to enhance osteoblast proliferation and formation of mineralized extracellular matrix.

A method has recently been developed for producing fibre-reinforced composites (FRC) with porous surfaces, intended for use as load-bearing orthopaedic implants. This study focuses on evaluation of the bone-bonding behaviour of FRC implants. Three types of cylindrical implants, i.e. FRC implants with a porous surface, solid polymethyl methacrylate (PMMA) implants and titanium (Ti) implants, were inserted in a transverse direction into the intercondular trabeculous bone area of distal femurs and proximal tibias of New Zealand White rabbits. Animals were sacrificed at 3, 6 and 12 weeks post operation, and push-out tests (n=5-6 per implant type per time point) were then carried out. At 12 weeks the shear force at the porous FRC-bone interface was significantly higher (283.3+/-55.3N) than the shear force at interfaces of solid PMMA/bone (14.4+/-11.0 N; p

Background Entropy™ is an anaesthetic EEG monitoring method, calculating two numerical parameters: State Entropy (SE, range 0–91) and Response Entropy (RE, range 0–100). Low Entropy numbers indicate unconsciousness. SE uses the frequency range 0.8–32 Hz, representing predominantly the EEG activity. RE is calculated at 0.8–47 Hz, consisting of both EEG and facial EMG. RE–SE difference (RE−SE) can indicate EMG, reflecting nociception. We studied RE−SE and EMG in patients anaesthetized without neuromuscular blockers. Methods Thirty-one women were studied in propofol–nitrous oxide (P) or propofol–nitrous oxide–remifentanil (PR) anaesthesia. Target SE value was 40–60. RE−SE was measured before and after endotracheal intubation, and before and after the commencement of surgery. The spectral content of the signal was analysed off-line. Appearance of EMG on EEG was verified visually. Results RE, SE, and RE−SE increased during intubation in both groups. Elevated RE was followed by increased SE values in most cases. In these patients, spectral analysis of the signal revealed increased activity starting from low (

In a study on 25 patients with verified benign bone tumors, bioactive glass (BG) and autogenous bone (AB) were used as bone-graft substitutes. The patients were randomized into two groups according to the filling material. Blood samples were taken both preoperatively, at 2 weeks, and 3, 8, 12, 24, and 36 months postoperatively, for evaluation of silicon concentration in blood. In the determination, direct current plasma atomic emission spectroscopy was used. No significant difference in blood silicon concentration between the BG group or the AB group could statistically be observed (p = 0.5400), and neither did the size of the bone tumor (p = 0.4259) nor the follow-up time affect the results (p = 0.2094). Concentration of osteocalcin in blood was significantly higher for large cysts (p < 0.0001). The filler material (BG or AB) did not affect the osteocalcin concentration level in blood.

In the coming decades, the need for reconstructive surgery of bones is predicted to increase with the ageing of the population as well as the increase of injuries needing traumatologic treatments. Therefore, there is still a constant search for tissue engineering and bone substitute materials. Xenografts, synthetic hydroxyapatitite, bioactive glasses and other bone substitutes have widely been studied. When bone defects are filled using bioceramics in granules, their utilization is limited to small size defects, because the injected granules do not give immediate support against the biomechanical loading of the bone. The aim of this study was to evaluate the preliminary biomineralization and the compression strength of experimental injectable bone cements modified with calcium ceramics. Our studies have focused on the development of injectable composites of bone cements, i.e. in situ curable resin systems containing impregnated Ca ceramics. The polymerized bone cement composites aspire to simulate as closely as possible the mechanical and structural properties properties of bone. The present compressive strength of our inorganic-organic bone cements are >65 up to ~180 MPa. These cements are slightly porous from their outermost surface and showed preliminarily osteoconductivity of some degree.

Polymer technology has provided solutions for filling of bone defects in situations where there may be technical or biological complications with autografts, allografts, and metal prostheses. We present an experimental study on segmental bone defect reconstruction using a polymethylmethacrylate-(PMMA-) based bulk polymer implant prosthesis. We concentrated on osteoconductivity and surface characteristics.A critical size segment defect of the rabbit tibia in 19 animals aged 18-24 weeks was reconstructed with a surface porous glass fiber-reinforced (SPF) prosthesis made of polymethylmethacrylate (PMMA). The biomechanical properties of SPF implant material were previously adjusted technically to mimic the properties of normal cortical bone. A plain PMMA implant with no porosity or fiber reinforcement was used as a control. Radiology, histomorphometry, and scanning electron microscopy (SEM) were used for analysis of bone growth into the prosthesis during incorporation.The radiographic and histological incorporation model showed good host bone contact, and strong formation of new bone as double cortex. Histomorphometric evaluation showed that the bone contact index (BCI) at the posterior surface interface was higher with the SPF implant than for the control. The total appositional bone growth over the posterior surface (area %) was also stronger for the SPF implant than for controls. Both bone growth into the porous surface and the BCI results were related to the quality, coverage, and regularity of the microstructure of the porous surface.Porous surface structure enhanced appositional bone growth onto the SPF implant. Under load-bearing conditions the implant appears to function like an osteoconductive prosthesis, which enables direct mobilization and rapid return to full weight bearing.

Biodegradable polymers (e.g. poly-ε-caprolactone, PCL) have been studied largely for tissue engineering applications. The aim of this study was to evaluate the composite fabrication technique on PCL modified with the phosphate salts (i.e. NaH2PO4, Na2HPO4, KH2PO4, or K2HPO4) as well as to determine the compression strengths thereof. The chemical structure and morphology of composites were analyzed using FTIR and SEM/EDX. The influence of a plain phosphate salt in different quantities on the hydrophilic properties of PCL was evaluated by measuring the water contact angle. The results of this study indicated that the addition of phosphate salts led to an improvement in compression strength of PCL composites. According to the results of preliminary biomimetic mineralization, Na2HPO4 seems to increase the bioactivity of PCL.

In this study, tissue reactions and possible toxicological responses of two different bioactive and degradable composite materials consisting of poly( ∈-caprolactone-co-DL-lactide) and bioactive glass (S53P4) granules were evaluated. The chosen materials were implanted subcutaneously in the back of rats for six months. The glass granules retained their bioactivity within the polymer matrices. A fibrous capsule formed around all tested materials and around the materials containing bioactive glass the fibrous capsules appeared to be thicker. Tissue growth into these materials was observed during the healing period while no growth was noticed into the plain polymer matrices. No adverse reactions were seen with any of the evaluated materials.

Medical polymers of biostable nature (e.g. polymethylmetacrylate, PMMA) are widely used in various clinical applications. In this study, novel PMMA-based composite bone cement was prepared. Bioactive glass (BAG) particulate filler (30wt%) was added to enhance potentially the integration of bone to the cement. The polymer matrix was functionalized with trimethoxysilyl to achieve an interfacial bond between the matrix and the fillers of BAG. The amount of trimethoxysilyl in the monomer system varied from 0 to 75wt%. The effects of dry and wet (simulated body fluid, SBF at +37°C for 5 weeks) conditions were investigated. In total, 20 groups of specimens were prepared. The specimens were subjected to a destructive mechanical test in compression. Scanning electron microscopy (SEM) and micro-computed tomography (micro-CT) were used to study the surface and the three-dimensional morphology of the specimens. The results of the study indicated that the addition of trimethoxysilyl groups led to the formation of a hybrid polymer matrix which, in lower amounts (

Background Concomitantly recorded Bispectral Index® (BIS) and Entropy™ values sometimes show discordant trends during general anaesthesia. Previously, no attempt had been made to discover which EEG characteristics cause discrepancies between BIS and Entropy. We compared BIS and Entropy values, and analysed the changes in the raw EEG signal during surgical anaesthesia with sevoflurane. Methods In this prospective, open-label study, 65 patients receiving general anaesthesia with sevoflurane were enrolled. BIS, Entropy and multichannel digital EEG were recorded. Concurrent BIS and State Entropy (SE) values were selected. Whenever BIS and SE values showed ≥10-unit disagreement for ≥60 s, the raw EEG signal was analysed both in time and frequency domain. Results A ≥10-unit disagreement ≥60 s was detected 428 times in 51 patients. These 428 episodes accounted for 5158 (11%) out of 45 918 analysed index pairs. During EEG burst suppression, SE was higher than BIS in 35 out of 49 episodes. During delta-theta dominance, BIS was higher than SE in 141 out of 157 episodes. During alpha or beta activity, SE was higher than BIS in all 49 episodes. During electrocautery, both BIS and SE changed, sometimes in the opposite direction, but returned to baseline values after electrocautery. Electromyography caused index disagreement four times (BIS > SE). Conclusions Certain specific EEG patterns, and artifacts, are associated with discrepancies between BIS and SE. Time and frequency domain analyses of the original EEG improve the interpretation of studies involving BIS, Entropy and other EEG-based indices. Clinical trial registration.ClinicalTrials.gov:identifier NCT01077674.

To investigate the role of pancreatic (group I) secretory PLA2 (sPLA2-I) in the pathogenesis of meconium aspiration syndrome, human particulate meconium or its supernatant either before or after extraction of PLA2-I was insufflated into rat lungs. In addition, the pulmonary effects of intra-tracheal human and bovine PLA2-I were studied. Lungs with saline instillation served as controls. Intrapulmonary particulate meconium (both before and after PLA2-I extraction), unlike meconium supernatant, resulted in markedly elevated lung tissue PLA2 catalytic activity and human PLA2-I concentrations when compared with controls. On the other hand, tissue concentrations of the group II PLA2 remained unchanged in all meconium lungs. Pulmonary PLA2-I concentrations further correlated positively with lung injury scores. Instillation of meconium-derived human PLA2-I, at a concentration of one-third of that in particulate meconium, did not raise PLA2 activity or concentrations of PLA2-I or PLA2-II in the lung tissue from the control level, but still resulted in significantly elevated lung wet/dry ratio and injury score. In contrast, insufflation of bovine pancreatic PLA2 increased the lung tissue enzyme activity and wet/dry ratio from the control level, but had no effect on the type II PLA2 concentration or lung injury score. Our data thus indicate that human pancreatic PLA2, introduced in high amounts within aspirated meconium especially in particulate form, is a potent inducer of lung tissue inflammatory injury.

Pulmonary inflammation and parenchymal apoptosis are implicated in the pathogenesis of the acute lung injury, but the mechanisms of these reactions are still unclear. Because inhibition of the proinflammatory cyclo-oxygenase (COX)-2 enzyme action is proposed to be useful in various inflammatory lung injuries, we decided to investigate the expression of COX-2 and the possible beneficial effects of its inhibition on pulmonary inflammation and apoptosis in surfactant-depleted lungs. The injury was induced in 2-mo-old rats by repeated lung lavage to remove alveolar surfactant. Eight of these rats were pretreated with a specific COX-2 inhibitor, NS-398. All rats, including control rats without lung lavage, were ventilated with 60% oxygen for 5 h, and the lungs were then studied histologically for tissue injury and with DNA nick-end labeling, cleaved caspase-3 immunohistochemistry, and electron microscopy for apoptotic cell death. Lung tissue myeloperoxidase activity and the expression of COX-2 protein and concentration of prostaglandin E2 were additionally analyzed. Lung lavage increased pulmonary neutrophil migration, histologic injury, and the occurrence of epithelial apoptosis. In contrast, expression of COX-2 and amount of PGE2 were significantly lower in surfactant-depleted lungs than controls. Pretreatment with the COX-2 inhibitor further increased the migration of neutrophils and occurrence of epithelial apoptosis in the surfactant-depleted lungs, compared with nontreated insulted lungs. These results suggest that specific inhibitors of COX-2 should be used cautiously in association with surfactant-deficient lung injuries.

Cutaneous neurofibromas consist of axonal processes, Schwann cells, fibroblasts, perineurial cells, mast cells, and abundant extracellular matrix. The distribution and role of perineurial cells in neurofibromas has been uncertain, partly because there has not been a specific immunohistochemical marker for perineurial cells. In this study, tight junctions (TJs) of 16 neurofibromas from 12 patients with neurofibromatosis type 1 (NF1) were analyzed using electron microscopy, immunohistochemistry, and Western transfer analysis. Cell-cell contacts with typical ultrastructural morphology of TJs were seen between adjacent perineurial cells surrounding the small nerves and between contacting perineurial cell processes embedded in tumor stroma. Immunohistochemistry showed expression of claudin-1, claudin-3, and ZO-1 in the intercellular junctions of a subpopulation of tumor cells. Occludin was present mainly in perineurium and claudin-5 localized to the blood vessels. Double immunolabelings were used to identify the cell types expressing claudin-1. The results showed that claudin-1 positive cells were also positive for type IV collagen and epithelial membrane antigen but not for S-100 protein. This labeling pattern is consistent with perineurial cell phenotype. Using claudin-1 as a marker, our results showed that clusters of perineurial cells are distributed around the rudimentary nerves within cutaneous neurofibromas and at the periphery of some neurofibromas.

We examined new bone formation after filling cancellous bone defects with bioactive glass (BG) in granular form. Cylindrical defects in the trochanter area of 18 rabbit femora were filled with BG granules (diameter 600-830 microns) and compared with similar defects filled with morcellized autogenous bone. New bone formation and surface reaction of BG particles were evaluated by light microscopy, histomorphometry, and scanning electron microscopy. The chemical profile at the bone- material interface was studied by energy dispersive x-ray analysis (EDXA). In the BG group, 41, 32, and 38 percent of the defects were filled with new bone after 3, 6, and 12 weeks, respectively. The corresponding figures for the autogenous bone group were 36, 29, and 34 percent. The thickness of the reaction layer on the glass surface increased from 82 to 163 microns during the observation periods. An intimate contact without intervening soft tissue between new bone lamellae and BG granules was a constant finding. EDXA showed a chemical continuum between the granules and the new bone. No adverse reactions related to BG were observed. BG is a promising material for filling cancellous bone defects.

To determine the expression of bactericidal/permeability-increasing protein (BPI), a novel antimicrobial molecule, in the main lacrimal gland and its content in tears of young healthy subjects.BPI concentration of tears was measured in 42 healthy volunteers, 13 men and 29 women, with ages ranging from 22 to 30 (mean 24.7+/-2.1) years by a time-resolved fluoroimmunoassay (TR-FIA). Immunohistochemical analysis was made to localize BPI in lacrimal gland and conjunctiva of eight autopsied subjects, two men and six women, with the age range from 44 to 87 (mean 72.3+/-14.9) years.The mean concentration of BPI in tears was 27.8+/-29.5 microg/l, and it decreased with an increase in tear flow rate (P0.0001). There was no statistically significant difference in BPI content of tears between the genders. BPI was immunohistochemically seen in outer basal epithelial cells of intralobular and excretory ducts, squamous and basal cells of conjunctiva as well as faintly in myoepithelial cell layer of acini. The presence of BPI in the lacrimal gland and in the tear fluid was verified by Western blotting.The results indicate that outer basal epithelial cells of lacrimal gland ducts contain BPI, which occurs in a relatively high concentration in tears. BPI may have a substantial antibacterial role in human tears.

Pneumocyte apoptosis is implicated in the pathophysiology of acute inflammatory lung injuries in newborns and adults. Pulmonary angiotensin (ANG) II contributes to lung epithelial apoptosis in vitro, but its role in acute lung injury in vivo is unclear. We therefore studied the effects of ANG II receptor action on the pulmonary inflammatory and apoptotic changes in surfactant-depleted lungs in rats. Lung injury was induced by repeated lung lavage with saline, and the rats were then ventilated with 60% oxygen for 1, 3, or 5 hr. Separate groups of rats were pretreated with a nonspecific ANG II receptor inhibitor saralasin, the specific ANG II type 1 receptor antagonist losartan, or ANG II type 2 receptor inhibitor PD123319, and were similarly studied. Lungs were studied histologically for tissue injury, and with terminal deoxynucleodityl transferase-mediated dUTP nick end-labeling (TUNEL) and cleaved caspase 3 antibody staining, and by electron microscopy for apoptotic cell death. Surfactant-depleted lungs showed an increased number of TUNEL-positive epithelial cells throughout the study, and intrapulmonary leukocyte migration and histological tissue injury scores were similarly elevated, compared to controls, from 1-5 hr of ventilation. Pretreatment with saralasin or losartan significantly prevented the increase of TUNEL positivity in pneumocytes, but had no effect on the amount of neutrophil influx or total injury score in lavaged lungs. In contrast, administration of PD123319 did not affect the number of TUNEL-positive epithelial cells or histological injury . The results suggest that increased epithelial apoptosis in surfactant-deficient lungs is mediated by ANG II receptor (specifically, subtype 1) action.

In Finland the question of appropriate wastewater treatment in rural areas has become very prominent during the last years. The new Environmental Protection Act stipulates that wastewaters must be treated to the extent that they cannot have a negative impact on nature. The Ministry of the Environment is currently preparing a decree specifying the requirements. The draft of the decree proposes that on-site treatment units should decrease the BOD load by 90, total phosphorus load by 85 and total nitrogen load by 40 per cent. To meet the new requirements, the old systems that include septic tanks only need more efficient wastewater treatment methods. Whatever technical solution is selected, the house owner must pay for it. At the moment, even the septic tanks are emptied and maintained irregularly. More sophisticated wastewater treatment methods definitely need more maintenance, which cannot be made the sole duty of the house owners. One potential organisational alternative for managing wastewater treatment in rural areas is the co-operative. Finland has one such pioneering co-operative formed for on-site sanitation. Varsinais-Suomi Water Services Co-operative provides the house owner with professional assistance in wastewater treatment at a reasonable cost. Suvisaari Water Services Co-operative is another new organisation selling sewerage services to its members. But its technology is different: this co-operative operates an LPS-sewerage system instead of on-site treatment.

The aim of this study was to describe and evaluate the significance of a porous surface with bioactive glass granules (S53P4) covering an artificial bulk material based on polymethylmetacrylate (PMMA) and fibre-reinforced composite (FRC) technology. Effort was focused particularly on characters of the porous surface and biomechanical properties of the material in vitro , and test in vivo the implant in reconstruction in an experimental long bone segment defect model. The defect, 10 mm in length, created in the shaft of rabbit tibia, was reconstructed by the implant and fixed by intramedullary K-wires. The implant was incorporated within 4 weeks by new bone growth from the host bone covering particularly its posterior surface and cortex/implant junctions with bridging trabecular bone. Later, at 8 weeks, new bone was found also at the cortex/implant interface and in the medullary canal of the implant. Histometric measurements revealed direct bone/implant surface contact in 34% at the interface. Bioactive glass granules in the porous surface evoked the most direct contact with bone. The implants manufactured from PMMA only served as a control group, and showed significantly lower osteoconductive properties. Biomechanical measurements in vitro of fibre-reinforced PMMA specimens revealed values for bending strength and the flexural modulus to match them to human bone. This artificial bulk bone material based on PMMA/FRC technology seems to have proposing properties to be used as a bone substitute on load-bearing conditions.

Lung tissue inflammation and apoptosis are implicated in the pathogenesis of meconium aspiration-induced lung injury in the newborn, but the mechanisms of these reactions are still poorly known. We investigated the time-dependent leukocyte influx and appearance of apoptosis, as well as the contribution of angiotensin (ANG) II receptor action on these processes in the meconium-induced lung injury. Experimental meconium aspiration was induced by intratracheal instillation of human meconium in 18 rats, and eight rats were further pretreated with an unspecific ANG II receptor inhibitor saralasin. Rats were ventilated with 60% oxygen for 1, 3, or 5 h, and the lungs were then studied histologically for tissue injury and with DNA nick-end labeling and electron microscopy for apoptotic cell death. Lung tissue myeloperoxidase activity and expression of angiotensinogen mRNA and endothelial monocyte-activating polypeptide (EMAP) II protein were also analyzed. The meconium-instilled lungs showed increasing neutrophil migration and histologic injury after the first hour, whereas the number of epithelial apoptotic cells was elevated from the control level throughout the study. Myeloperoxidase activity was high, and the angiotensinogen mRNA and EMAP II protein was up-regulated at 5 h after the meconium insult. Pretreatment with saralasin significantly prevented the increase in lung tissue myeloperoxidase activity, EMAP II, and lung epithelial apoptosis. The results suggest that pulmonary meconium insult rapidly results in epithelial apoptosis, before significant neutrophil sequestration into the lungs. Apoptotic cell death is further connected with ANG II receptor action in the meconium-contaminated lung tissue.

Neonatal meconium aspiration often produces severe respiratory distress due to an inflammatory pulmonary injury, but the extension of this damaging reaction to the noncontaminated lung regions is still uncertain. To investigate the presence of generalized pulmonary inflammatory response, 31 anesthetized and ventilated neonatal piglets (1-3 d) were studied. Meconium (n = 16) or saline (n = 15) was instilled unilaterally into the right lung, and analysis of the lung tissue or bronchoalveolar lavage (BAL) fluid from both lungs was performed after 12 h. Meconium increased the wet/dry weight ratio, histologic tissue injury score and tissue myeloperoxidase activity as well as BAL fluid total cell count in the contaminated lung. Tumor necrosis factor-alfa concentrations in BAL fluid did not however differ significantly. Furthermore, in the meconium-instilled lungs the tissue and lavage fluid catalytic activity of phospholipase A2 (PLA2) and tissue PLA2 group-I and group-II concentrations were significantly elevated. Although BAL fluid catalytic activity of PLA2 was moderately increased also in the meconium noninstilled lung, significant inflammatory injury in this lung was absent. The results thus indicate that meconium aspiration induces severe local inflammation and lung injury, but significant generalized pulmonary inflammatory damage in the pathogenesis of meconium aspiration syndrome is unlikely.

Bioactive glass granules of three different compositions, regarding particularly Si- and Al- content (S53P4, S59.7P2.5, S52P3) and of two different granule sizes (200-250 microm and 630-800 microm) were implanted for 4 and 8 weeks in the distal part of rabbit femur. The effect of glass composition and granule size on bone formation was studied. The results were evaluated using histology, computerized histomorphometry, scanning electron microscopy and energy dispersive X-ray analysis, and used for mathematical description of bone formation. The results showed that both the composition of the glass and the granule size of the granules, have influence on bone growth from the surrounding tissue. Glass S53P4, which from previous observations is known to be an effective bioactive glass and widely used in the Biomaterial Project of Turku, Finland, showed bone bonding and increasing bone growth between the granules. Glass S59.7P2.5 which due to its high Si-content should be inert, showed bone bonding. At 4 weeks the bone growth was significantly more abundant in bone defects filled with large granules (630-800 microm) than in defects filled with small granules (200-250 microm). Glass S52P3 with an alumina content of 3 wt %, showed good bone conduction, possibly even bone bonding for granules of 630-800 microm size. Granules of 200-250 microm with a high alumina content at the surface of the reaction layer, showed hardly any bone contact at all. This data, therefore, gives new information concerning bone bonding and osteoconduction of bioactive glasses with a high silica or alumina content.