Cultivation in rotating bioreactors promotes maintenance of cardiac myocyte electrophysiology and molecular properties

We tested the hypothesis that cardiomyocytes maintained their phenotype better if cultured as three-dimensional tissue constructs than if cultured as confluent monolayers. Neonatal rat cardiomyocytes were cultured on biomaterial scaffolds in rotating bioreactors for 1 week, and resulting tissue constructs were compared with confluent monolayers and slices of native ventricular tissue with respect to proteins involved in cell metabolism (creatine kinase isoform MM), contractile function (sarcomeric myosin heavy chain), and intercellular communication (connexin 43), as well as action potential characteristics (e.g., membrane resting potential, maximum depolarization slope, and action potential duration), and macroscopic electrophysiological properties (maximum capture rate). The molecular and electrophysiological properties of cardiomyocytes cultured in tissue constructs, although inferior to those of native neonatal ventricles, were superior to those of the same cells cultured as monolayers. Construct levels of creatine kinase, myosin heavy chain, and connexin 43 were 40-60% as high as ventricle levels, whereas monolayer levels of the same proteins were only 11-20% as high. Construct action potential durations were 1.8-fold higher than those in ventricles, whereas monolayer action potential durations were 2.4-fold higher. Pharmacological studies using 4-aminopyridine showed that prolonged action potential duration and reduced maximum capture rate in tissue constructs as compared with native ventricles could be explained by decreased transient outward potassium current.     

Vascular thoracic outlet syndrome

Abstract The surgical treatment of 30 cases of vascular thoracic outlet syndrome (TOS) in 25 patients is presented. Patients included 17 women and 8 men with average age of 26.1 years. The causes of compression were cervical rib (n = 16), soft tissue anomalies (n = 12), and scar tissue after clavicle fracture (n = 2). Ten subclavian artery aneurysms containing intraluminal thrombus as well as one subclavian artery occlusion were found. All such cases had multiple distal arterial embolization. Presenting features of cases with arterial TOS included: hand ischemia (n = 11), transient ischemic attack (TIA) (n = 1), and claudication or vasomotor phenomena during the arm hyperabduction (n = 11). Two patients with venous TOS developed hand edema during arm hyperabduction, and five other patients had axillary-subclavian venous thrombosis. In all cases decompressive procedures using a combined supraclavicular and infraclavicular approach were performed. Decompression was achieved by cervical rib excision (n = 12), combined cervical and first rib excision (n = 4), and first rib excision (n = 14). In all cases division of all soft tissue elements was also accomplished. Associated vascular procedures included resection and replacement of 10 subclavian artery aneurysms, one subclavian-axillary and one axillary-brachial bypass, as well as nine brachial embolectomies. All five cases with axillary-subclavian vein thrombosis before decompression were treated with anticoagulant therapy. The mean follow-up period was 3 years and 2 months (range 1 to 6 years). Two pleural entry injuries and two transient brachial plexus injuries were noted. All reconstructed arteries were patent during the follow-up period. Complete resolution of symptoms with a return to full activity was noticed in all cases with arterial TOS and in two cases with venous TOS without axillary-subclavian vein thrombosis. In cases with axillary-subclavian vein thrombosis relief of symptoms was mild, and there were limitations on daily activity. Vascular TOS is seen less frequently than the neurogenic form; however, in most cases it requires surgical treatment. We prefer a combined supraclavicular and infraclavicular approach because it offers complete exposure of the subclavian artery, cervical and first ribs, and all soft tissue anomalies. Electronic Publication     

WHO's ICF and functional status information in health records

A common framework for describing functional status information (FSI) in health records is needed in order to make this information comparable and of value. The World Health Organization's (WHO's) International Classification of Functioning, Disability and Health (ICF), which has been approved by all its member States, provides this common language and framework. The biopsychosocial model of functioning and disability embodied in the ICF goes beyond disease and conceptualizes functioning from the individual's body, person, and lived experience vantage points, thereby allowing for planning interventions targeted at the individual's body, the individual as a whole or toward the environment. This framework then permits the evaluation of both the effectiveness and cost effectiveness of these different interventions in devising programs at the personal or societal level.     

Auditory peripheral influences on calcium binding protein immunoreactivity in the cochlear nucleus during aging in the C57BL/6J mouse

The C57BL/6J (C57) mouse was selected as a suitable model for early presbyacusis to determine if there were correlations between peripheral pathology (spiral ganglion loss, inner and outer hair cell loss) and calcium binding immunoreactivity in the cochlear nucleus during aging. The quantitative stereological method, the optical fractionator, was used for determining the total number of neurons and calcium binding immunopositive neurons (calbindin, parvalbumin and calretinin) during aging in the posteroventral- and dorsal cochlear nucleus (PVCN and DCN) in C57 mice. Comparing 30-month-old to 1-month-old C57 mice, a percent increase in parvalbumin and calbindin immunoreactivity was evident in both the PVCN and DCN. Correlations were made between peripheral pathology (spiral ganglion and inner and outer hair cell loss) and calcium binding protein expression. Significant correlations between cochlear pathology and the percentage of parvalbumin and calretinin immunoreactive neurons were demonstrated in the DCN. Moreover, significant correlations were found between cochlear pathology and parvalbumin and calbindin in the PVCN. In summary, the findings imply that degenerative changes in the auditory periphery can modulate neuronal homeostasis by increasing calcium binding proteins in the PVCN and DCN during aging. Taken together, these findings suggest a role for calcium binding proteins in protecting against age-induced calcium toxicity.     

Advantages and limitations of the autofluorescent diagnostics of the laryngeal cancer and precancerosis

Any endoscopic diagnostic procedure that is capable of giving exact information on laryngeal lesions without damaging the tissue has essential advantages over standard biopsy. Tissue autofluorescence is defined as a natural ability of tissue to fluoresce when exposed to a certain light wavelength. This feature is a consequence of the presence of fluorophores in the tissues, which are activated by a narrow wavelength range. However, due to their biochemical and biophysical characteristics, laryngeal precancerosis and cancer do not fluoresce when exposed to blue light. In the present study, we used Pentax’s System of Autofluorescent Endoscopy (SAFE 1000) to detect autofluorescence disturbances from laryngeal mucosa. Diagnostic parameters (sensitivity and specificity) of the microlaryngoscopy (MLS) and SAFE 1000 in the diagnosis of laryngeal precancerosis and carcinoma were compared and discussed. We have found that SAFE had a better sensitivity with regard to mentioned laryngeal pathology, but MLS had better specificity than SAFE. The overall diagnostic sensitivity in the diagnostics of laryngeal atypical hyperplasia and cancer with SAFE was 89%, as opposed to 73% with MLS. Diagnostic specificity of SAFE for all cases of laryngeal carcinomas and atypical hyperplasia was 78%. The specificity of MLS in diagnostics of laryngeal carcinomas cases was 98%, while that for cases of atypical hyperplasia was 100%. Many other conditions that have impact on autofluorescent features of laryngeal mucosa were also discussed.     

Fetal and maternal plasma homocysteine levels during the second half of uncomplicated pregnancy

Objective: To measure fetal and maternal plasma homocysteine (Hcy) concentrations in uncomplicated pregnancies.

Methods: Paired maternal venous and fetal umbilical cord blood (n?=?81) samples were evaluated for plasma Hcy and vitamin B₁₂ levels, in addition to eight neonatal umbilical cord blood samples obtained immediately following delivery.

Results: Both fetal and maternal Hcy concentrations were positively correlated with advancing gestational age (ρ?=?0.44, p?<?0.0001; and ρ?=?0.27, p?<?0.05, respectively). Fetal plasma Hcy concentrations [2.2?µmol/l (IQR: 2.0–3.2)] were significantly lower than both neonatal umbilical vein [5.0?µmol/l (IQR: 4.4–6.5); p?<?0.001] and maternal plasma Hcy levels [4.4?μmo/l (IQR: 3.4–5.4); p?<?0.001]. In addition, Hcy values at term were higher in the umbilical vein compared with the umbilical artery [5.0?μmol/l (IQR: 3.4–5.4) versus 4.2?μmol/l (IQR: 3.7–5.5), respectively; p?=?0.016]. Significant correlation was noted and between fetal and maternal Hcy levels (ρ?=?0.50, p?<?0.0001), while fetal Hcy was negatively correlated with maternal B₁₂ concentrations (ρ?=??0.32, p?<?0.001).

Conclusions: Fetal Hcy levels were significantly lower than maternal and neonatal levels and correlated with gestational age across the second half of pregnancy.     

Procalcitonin in preoperative diagnosis of abdominal sepsis

Background and aims The present study attempted to identify the diagnostic significance of procalcitonin (PCT) in acute abdominal conditions as well as the range of concentrations relating to diagnosis of abdominal sepsis. Materials and methods This was prospective clinical study. The study included 98 consecutive patients with acute abdominal conditions, divided in sepsis and systemic inflammatory response syndrome (SIRS) group. Results PCT concentrations on admission were significantly higher in the sepsis group than in the SIRS group (median [interquartile range] 2.32 [7.41] vs 0.45 ng/ml [2.62]). A cutoff value of 1.1 ng/ml yielded 72.4% sensitivity and 62.5% specificity. In a group of patients with abdominal symptoms lasting for more than 24 h, a cut-off value of 1.1 ng/ml yielded higher sensitivity (82.9%) and higher specificity (77.3%). Conclusion Our results suggest that PCT measurements may be useful for early, preoperative diagnosis of abdominal sepsis.     

EAU Guidelines on Urethral Trauma

Context

These guidelines were prepared on behalf of the European Association of Urology (EAU) to assist urologists in the management of traumatic urethral injuries.

Objective

To determine the optimal evaluation and management of urethral injuries by review of the world's literature on the subject.

Evidence acquisition

A working group of experts on Urological Trauma was convened to review and summarize the literature concerning the diagnosis and treatment of genitourinary trauma, including urethral trauma. The Urological Trauma guidelines have been based on a review of the literature identified using on-line searches of MEDLINE and other source documents published before 2009. A critical assessment of the findings was made, not involving a formal appraisal of the data. There were few high-powered, randomized, controlled trials in this area and considerable available data was provided by retrospective studies. The Working Group recognizes this limitation.

Evidence synthesis

The full text of these guidelines is available through the EAU Central Office and the EAU website (www.uroweb.org). This article comprises the abridged version of a section of the Urological Trauma guidelines.

Conclusions

Updated and critically reviewed Guidelines on Urethral Trauma are presented. The aim of these guidelines is to provide support to the practicing urologist since urethral injuries carry substantial morbidity. The diversity of urethral injuries, associated injuries, the timing and availability of treatment options as well as their relative rarity contribute to the controversies in the management of urethral trauma.     

Pluripotent stem cell-derived cardiac tissue patch with advanced structure and function

Recent advances in pluripotent stem cell research have provided investigators with potent sources of cardiogenic cells. However, tissue engineering methodologies to assemble cardiac progenitors into aligned, 3-dimensional (3D) myocardial tissues capable of physiologically relevant electrical conduction and force generation are lacking. In this study, we introduced 3D cell alignment cues in a fibrin-based hydrogel matrix to engineer highly functional cardiac tissues from genetically purified mouse embryonic stem cell-derived cardiomyocytes (CMs) and cardiovascular progenitors (CVPs). Procedures for CM and CVP derivation, purification, and functional differentiation in monolayer cultures were first optimized to yield robust intercellular coupling and maximize velocity of action potential propagation. A versatile soft-lithography technique was then applied to reproducibly fabricate engineered cardiac tissues with controllable size and 3D architecture. While purified CMs assembled into a functional 3D syncytium only when supplemented with supporting non-myocytes, purified CVPs differentiated into cardiomyocytes, smooth muscle, and endothelial cells, and autonomously supported the formation of functional cardiac tissues. After a total culture time similar to period of mouse embryonic development (21 days), the engineered cardiac tissues exhibited unprecedented levels of 3D organization and functional differentiation characteristic of native neonatal myocardium, including: 1) dense, uniformly aligned, highly differentiated and electromechanically coupled cardiomyocytes, 2) rapid action potential conduction with velocities between 22 and 25 cm/s, and 3) significant contractile forces of up to 2 mN. These results represent an important advancement in stem cell-based cardiac tissue engineering and provide the foundation for exploiting the exciting progress in pluripotent stem cell research in the future tissue engineering therapies for heart disease.     

Cancer epidemiology in Central, South and Eastern European countries

Aim

To collect cancer epidemiology data in South Eastern European countries as a basis for potential comparison of their performance in cancer care.

Methods

The South Eastern European Research Oncology Group (SEEROG) collected and analyzed epidemiological data on incidence and mortality that reflect cancer management in 8 countries – Croatia, Czech Republic, Hungary, Romania, Poland, Slovakia, and Serbia and Montenegro in the last 20-40 years.

Results

The most common cancer type in men in all countries was lung cancer, followed by colorectal and prostate cancer, with the exception of the Czech Republic, where prostate cancer and colorectal cancer were more common. The most frequent cancer in women was breast cancer followed by colorectal cancer, with the exceptions of Romania and Central Serbia where cervical cancer was the second most common. Cancer mortality data from the last 20-40 years revealed two different patterns in men. In Romania and in Serbia and Montenegro, there was a trend toward an increase, while in the other countries mortality was declining, after increasing for a number of years. In women, a steady decline was observed over many years in the Czech Republic, Hungary, and Slovakia, while in the other countries it remained unchanged.

Conclusions

There are striking variations in the risk of different cancers by geographic area. Most of the international variation is due to exposure to known or suspected risk factors which provides a clear challenge to prevention. There are some differences in incidence and mortality that cannot be explained by exposure to known risk factors or treatment availabilities.On a global scale, cancer has become a major public health problem and an increasingly important contributor to the burden of disease. Based on the most recent available international data, there were an estimated 12.7 million new cancer cases, 7.6 million deaths from cancer, and 28 million persons alive with cancer within five years from the initial diagnosis (1-3). The most common cancers in the world were lung (1.61 million cases), breast (1.38 million), and colorectal cancer (1.24 million) (3). Because of its poor prognosis, lung cancer was also the most common cause of death (1.38 million), followed by gastric (737 000 deaths), and liver cancer (695 000 deaths) (1-4).Priority setting for cancer control and cancer services in any region needs to be based on knowledge of the cancer burden and the local mix of predominant cancer types (5). According to estimates of global cancer burden made by the International Agency for Research on Cancer (IARC), the incidence and mortality rates from many specific types of cancer and all cancers combined vary widely by geographic locality (6). Moreover, the IARC also estimated that over half of newly diagnosed cases and two-thirds of cancer deaths occur in low and medium-income countries (6). There are striking variations in the pattern of cancer by site from region to region (7). The large differences in incidence and mortality in different countries may reflect a combination of differences in prevalence of underlying risk factors, differences in host susceptibility, and/or variations in cancer detection, reporting, classification systems, treatment, and follow-up. Among European countries, wide differences in the quality of cancer care are observed, especially when comparison is made between “old” and “new” EU members or between developed and developing countries (8). Cancer survival is significantly lower in Eastern European countries, including the new Member States, than in the EU 15 (9-12). Transitional countries and middle income countries are frequently left forgotten “in between” and the cancer problem in these countries is among the worst and fastest growing (8).In this report, we provide an analysis, which we propose as a foundation for detailed evaluation of cancer care in selected Central, Southern and Eastern European countries, represented by members of the South Eastern European Research Oncology Group (SEEROG). Our epidemiological analysis indicates the scale of the problem of oncological care in individual countries and shows current trends in the incidence of particular cancers. Comparison of status of oncology between countries in Eastern, Southern, and Central European region has never been undertaken before and key barriers to deliver appropriate quality of care have not previously been identified.