Giving blood and enrolling on the stem cell donor registry: ranking of obstacles and motives in Switzerland.

BACKGROUND: To obtain a better understanding of factors affecting blood and blood stem cell donation behavior in Switzerland, a series of studies has been performed. In the recent study of this series, which is described here, motivators and barriers in the field of blood and blood stem cell donation were identified. METHODS: Web-based survey data from a non-random sample of the Swiss population 2012/2013 (n = 3,153) were used to describe and compare the ranking of motives and obstacles to donate blood and to enroll on the Swiss blood stem cell registry. Wilcoxon rank-sum test and Spearman's rank correlations were used to assess differences and associations between ranks and groups. RESULTS: The prospect of saving lives and solidarity were the top two motives to donate blood or to enroll on the blood stem cell registry. The top two obstacles to enroll on the blood stem cell registry were lack of general information on blood stem cell donation and on its risks, whereas the top two obstacles to donate blood were the lack of information where and when to donate and deferral of or exclusion from blood donation. CONCLUSION: Classical altruistic motives are top drivers for giving blood as well as registering for blood stem cell donation. Recruitment campaigns should focus on these motivators. Similarities in motivational factors as well as in obstacles regarding blood and blood stem cell donation can be found.

Individual Characteristics Associated with Blood Donation: A Cross-National Comparison of the German and Swiss Population between 1994 and 2010.

OBJECTIVE: The aim of this study is to compare individual characteristics associated with blood donation in the German and Swiss population between 1994 and 2010. METHODS: Population-based survey data from the Eurobarometer 1994 and 2009, the Swiss Health Survey 1997, and the Swiss Blood Donation Survey 2010 were used to compare age-adjusted percentages of German and Swiss adults ever having donated blood (n = 8,746). A multivariate logistic regression was applied to the pooled data to estimate odds ratios (OR). RESULTS: Donor rates between 1994 and 2010 increased by 8.6% in Germany (p = 0.0045) and remained stable in Switzerland. The likelihood to report donating increased with age. Gender differences (OR = 2.85; p = 0.0000) and differences between education levels were more pronounced in Switzerland as compared to Germany (OR = 2.56; p = 0.0000 and OR = 2.73; p = 0.0010). Furthermore education differences were more marked in men in both countries (OR = 1.99; p = 0.0000 and OR = 1.68; p = 0.0140). CONCLUSION: The blood establishments should intensify their efforts to motivate women and lower educated people to give blood. Our data suggest that population-based surveys could be a helpful tool to describe donor rates in different countries and to guide future recruitment strategies.

Impact of selection of cord blood units from the United States and swiss registries on the cost of banking operations.

BACKGROUND: Over the last 2 decades, cord blood (CB) has become an important source of blood stem cells. Clinical experience has shown that CB is a viable source for blood stem cells in the field of unrelated hematopoietic blood stem cell transplantation. METHODS: Studies of CB units (CBUs) stored and ordered from the US (National Marrow Donor Program (NMDP) and Swiss (Swiss Blood Stem Cells (SBSQ)) CB registries were conducted to assess whether these CBUs met the needs of transplantation patients, as evidenced by units being selected for transplantation. These data were compared to international banking and selection data (Bone Marrow Donors Worldwide (BMDW), World Marrow Donor Association (WMDA)). Further analysis was conducted on whether current CB banking practices were economically viable given the units being selected from the registries for transplant. It should be mentioned that our analysis focused on usage, deliberately omitting any information about clinical outcomes of CB transplantation. RESULTS: A disproportionate number of units with high total nucleated cell (TNC) counts are selected, compared to the distribution of units by TNC available. Therefore, the decision to use a low threshold for banking purposes cannot be supported by economic analysis and may limit the economic viability of future public CB banking. CONCLUSIONS: We suggest significantly raising the TNC level used to determine a bankable unit. A level of 125 × 10(7) TNCs, maybe even 150 × 10(7) TNCs, might be a viable banking threshold. This would improve the return on inventory investments while meeting transplantation needs based on current selection criteria.

Stem cells in translation: impression of the ISSCR Regional Meeting in Florence.

The International Society for Stem Cell Research hosted an exciting meeting on stem cell-based translational medicine in Florence, Italy in September 2013. This report gives an overview of recent advances and breakthroughs presented at the meeting.

Diagnostic accuracy of the Rivalta test for feline infectious peritonitis.

BACKGROUND: The Rivalta test has been used routinely in Europe to diagnose feline infectious peritonitis (FIP) in cats with effusions, but its diagnostic accuracy is uncertain. OBJECTIVES: The objectives of this study were to calculate sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values of the Rivalta test for FIP and to identify correlations between a positive Rivalta test and variables measured in effusion fluid and peripheral blood. METHODS: In this retrospective study, medical records of cats with effusions were reviewed, and cats with conclusive results for the Rivalta test were included. The prevalence of FIP in this population was determined, and sensitivity, specificity, and PPV and NPV of the Rivalta test were calculated. Variables measured in effusion fluid and peripheral blood were compared between cats that had positive or negative Rivalta tests using the Mann-Whitney U-test and multivariate analysis. RESULTS: Of 851 cats with effusions, 782 had conclusively positive or negative results for the Rivalta test. A definitive final diagnosis was made in 497 of these cats. Prevalence of FIP in cats with effusion and a conclusive Rivalta test result was 34.6%. The Rivalta test had a sensitivity of 91.3%, specificity of 65.5%, PPV of 58.4%, and NPV of 93.4% for the diagnosis of FIP. These values increased when cats with lymphoma or bacterial infections were excluded, or when only cats ≤ 2 years were considered. Increased effusion cholesterol concentration and specific gravity as well as decreased serum albumin:globulin ratio and hyperbilirubinemia were positively correlated with positive Rivalta test results. CONCLUSIONS: Sensitivity, specificity, and PPV of the Rivalta test for the diagnosis of FIP were lower than previously reported except when used in young cats. The components in effusions that lead to a positive Rivalta test remain unknown, but the positivity is not simply related to high total protein concentration.

[Pericardial effusion in a cat with feline infectious peritonitis]. / Perikarderguss bei einer Katze mit feliner infektiöser Peritonitis.

This case report describes the disease progression of a male cat with pericardial effusion. Clinical signs (dyspnea, lethargy, and weakness) started very acutely. The initial laboratory profile showed only an increase in alanine aminotransferase enzyme activity. Diagnostic imaging revealed pericardial effusion. Effusion analysis showed a Rivalta-positive, modified transudate. Detection of feline coronavirus antigen in macrophages was negative. General condition and laboratory parameters dramatically worsened within seven days. Therefore, the owners decided to euthanize the cat. Even if effusion variables are macroscopically and microscopically suspicious for FIP, a definitive diagnosis of FIP could only be made by histology (including immunhistochemical staining).

Quantitative comparison of constitutive promoters in human ES cells.

BACKGROUND: Constitutive promoters that ensure sustained and high level gene expression are basic research tools that have a wide range of applications, including studies of human embryology and drug discovery in human embryonic stem cells (hESCs). Numerous cellular/viral promoters that ensure sustained gene expression in various cell types have been identified but systematic comparison of their activities in hESCs is still lacking. METHODOLOGY/PRINCIPAL FINDINGS: We have quantitatively compared promoter activities of five commonly used constitutive promoters, including the human ß-actin promoter (ACTB), cytomegalovirus (CMV), elongation factor-1α, (EF1α), phosphoglycerate kinase (PGK) and ubiquitinC (UbC) in hESCs. Lentiviral gene transfer was used to ensure stable integration of promoter-eGFP constructs into the hESCs genome. Promoter activities were quantitatively compared in long term culture of undifferentiated hESCs and in their differentiated progenies. CONCLUSION/SIGNIFICANCE: The ACTB, EF1α and PGK promoters showed stable activities during long term culture of undifferentiated hESCs. The ACTB promoter was superior by maintaining expression in 75-80% of the cells after 50 days in culture. During embryoid body (EB) differentiation, promoter activities of all five promoters decreased. Although the EF1α promoter was downregulated in approximately 50% of the cells, it was the most stable promoter during differentiation. Gene expression analysis of differentiated eGFP+ and eGFP- cells indicate that promoter activities might be restricted to specific cell lineages, suggesting the need to carefully select optimal promoters for constitutive gene expression in differentiated hESCs.

NANOG reporter cell lines generated by gene targeting in human embryonic stem cells.

BACKGROUND: Pluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role in maintaining hESC pluripotency, but the precise role and regulation of NANOG are not well defined. METHODOLOGY/PRINCIPAL FINDINGS: To facilitate the study of NANOG expression and regulation in viable hESC cultures, we generated fluorescent NANOG reporter cell lines by gene targeting in hESCs. In these reporter lines, the fluorescent reporter gene was co-expressed with endogenous NANOG and responded to experimental induction or repression of the NANOG promoter with appropriate changes in expression levels. Furthermore, NANOG reporter lines facilitated the separation of hESC populations based on NANOG expression levels and their subsequent characterization. Gene expression arrays on isolated hESC subpopulations revealed genes with differential expression in NANOG(high) and NANOG(low) hESCs, providing candidates for NANOG downstream targets hESCs. CONCLUSION/SIGNIFICANCE: The newly derived NANOG reporter hESC lines present novel tools to visualize NANOG expression in viable hESCs. In future applications, these reporter lines can be used to elucidate the function and regulation of NANOG in pluripotent hESCs.

Effects of nasal mask leak and heated humidification on nasal mucosa in the therapy with nasal continuous positive airway pressure (nCPAP).

The purpose of this study was to determine the objective short-term influence of nasal continuous positive airway pressure (nCPAP) therapy, nasal mask leak (NML) and heated humidifiers (HH) to nasal conditioning of spontaneously breathing subjects. This was a prospective, non-randomized, non-blinded day-time study. Eighteen healthy subjects were enrolled in the study. All subjects received nCPAP therapy for 60 min in three different conditions successively: (1) nCPAP without humidification, (2) nCPAP with a defined leakage of nasal mask (slashed circle 28.3 mm2) and (3) nCPAP with HH. Nasal humidity and temperature were measured in the anterior turbinate area using a miniaturized thermocouple and a relative humidity sensor. The measurements were accomplished at the beginning of therapy, after 60, 120 and 180 min. Absolute humidity (aH) in the anterior turbinate area decreased significantly (p = 0.0075) from 17.41 +/- 3.81 mg/l (baseline) to 15.27 +/- 2.21 mg/l (nCPAP alone). With attachment of a NML, aH decreased from 15.27 mg/l not significantly (p = 0.058) to 13.77 +/- 2.28 mg/l (nCPAP and NML) compared to nCPAP alone. After addition of heated humidification to nCPAP, aH increased again from 13.77 mg/l significantly (p = 0.042) to 15.29 +/- 3.51 mg/l (nCPAP and HH) compared to aH (nCPAP+NML). No difference was found between aH (nCPAP and HH) and aH (nCPAP alone). Airway temperature did not change significantly after application of nCPAP alone, nCPAP and NML, and nCPAP and HH. These data indicate that nCPAP therapy with NML tends to have more remarkable reduction of the nasal humidity than nCPAP therapy without NML. nCPAP with heated humidifier is able to compensate the dehydration effects induced by nCPAP therapy with NML by increasing the aH at the anterior turbinate area to the levels observed during breathing with nCPAP alone.

Normal brain cells contribute to the bystander effect in suicide gene therapy of malignant glioma.

PURPOSE: Lentiviral vectors pseudotyped with glycoproteins of the lymphocytic choriomeningitis virus (LCMV-GP) are promising candidates for gene therapy of malignant glioma, as they specifically and efficiently transduce glioma cells in vitro and in vivo. Here, we evaluated the therapeutic efficacy of LCMV-GP and vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped vectors. EXPERIMENTAL DESIGN: Therapeutic efficacy was tested for unmodified (9L) and DsRed-modified (9LDsRed) gliomas using the suicide gene thymidine kinase of the herpes simplex virus type 1 (HSV-1-tk). Positron emission tomography (PET) and magnetic resonance imaging were done to analyze transduction of tumors and monitor therapeutic outcome. RESULTS: LCMV-GP pseudotypes mediated a successful eradication of 9LDsRed tumors with 100% of long-term survivors. Before initiation of ganciclovir treatment, a strong HSV-1-tk expression within the tumor was detected by noninvasive PET using the tracer 9-[4-[(18)F]fluoro-3-(hydroxymethyl)butyl]guanine. Therapeutic outcome was successfully monitored by magnetic resonance imaging and PET imaging and correlated with the histopathologic data. In the 9L model, LCMV-GP and VSV-G pseudotyped lentiviral vectors displayed similar therapeutic efficacy. Further studies revealed that normal brain cells transduced with VSV-G pseudotypes were not eliminated by ganciclovir treatment and contributed significantly to the bystander killing of tumor cells. CONCLUSIONS: Suicide gene transfer using pseudotyped lentiviral vectors was very effective in the treatment of rat glioma and therefore is an attractive therapeutic strategy also in human glioblastoma especially in conjunction with an imaging-guided approach. In addition, high selectivity of gene transfer to tumor cells may not always be desirable for therapeutic genes that exert a clear bystander effect.

A retroviral packaging cell line for pseudotype vectors based on glioma-infiltrating progenitor cells.

BACKGROUND: Early clinical trials for gene therapy of human gliomas with retroviral packaging cells (PC) have been hampered by low transduction efficacy and lack of dissemination of PC within the tumor. In the current approach, these issues have been addressed by creating a stable packaging cell line for retroviral vectors pseudotyped with glycoproteins of lymphocytic choriomeningitis virus (LCMV) based on tumor-infiltrating progenitor cells. METHODS: Tumor-infiltrating progenitor cells, which had been isolated from adult rat bone marrow (BM-TIC), were modified to stably express Gag-Pol proteins of moloney murine leukemia virus (Mo-MLV) and glycoproteins of LCMV. Packaging of a retroviral vector was measured by titration experiments on human fibroblast cells as well as on mouse and human glioma cell lines. Additionally, gene transfer was tested in a rat glioma model in vivo. RESULTS: The BM-TIC-derived packaging cell line (BM-TIPC) produced retroviral vectors with titers between 2-8 x 10(3) transducing units (TU)/ml. Extended culturing of BM-TIPC over several weeks and freezing/thawing of cells did not affect vector titers. No replication-competent retrovirus was released from BM-TIPC. In a rat glioma model, BM-TIPC infiltrated the tumors extensively and with high specificity. Moreover, BM-TIPC mediated transduction of glioma cells in vivo. CONCLUSION: This proof-of-principle study shows that primary adult progenitor cells with tumor-infiltrating capacity can be genetically modified to stably produce retroviral LCMV pseudotype vectors. These BM-TIPC may be a useful tool to enhance specificity and efficacy of gene transfer to gliomas in patients.

Bystander killing of malignant glioma by bone marrow-derived tumor-infiltrating progenitor cells expressing a suicide gene.

Adult stem cells are promising cellular vehicles for therapy of malignant gliomas as they have the ability to migrate into these tumors and even track infiltrating tumor cells. However, their clinical use is limited by a low passaging capacity that impedes large-scale production. In the present study, a bone marrow-derived, highly proliferative subpopulation of mesenchymal stem cells (MSCs)-here termed bone marrow-derived tumor-infiltrating cells (BM-TICs)-was genetically modified for the treatment of malignant glioma. Upon injection into the tumor or the vicinity of the tumor, BM-TICs infiltrated solid parts as well as the border of rat 9L glioma. After intra-tumoral injection, BM-TICs expressing the thymidine kinase of herpes simplex virus (HSV-tk) and enhanced green fluorescent protein (BM-TIC-tk-GFP) were detected by non-invasive positron emission tomography (PET) using the tracer 9-[4-[(18)F]fluoro-3-hydroxymethyl)butyl]guanine ([(18)F]FHBG). A therapeutic effect was demonstrated in vitro and in vivo by BM-TICs expressing HSV-tk through bystander-mediated glioma cell killing. Therapeutic efficacy was monitored by PET as well as by magnetic resonance imaging (MRI) and strongly correlated with histological analysis. In conclusion, BM-TICs expressing a suicide gene were highly effective in the treatment of malignant glioma in a rat model and therefore hold great potential for the therapy of malignant brain tumors in humans.

Radiofrequency-induced thermotherapy on nasal polyps: preliminary results.

BACKGROUND: The aim of this study was to determine the effects of radiofrequency-induced thermotherapy (RFITT) in patients with nasal polyps. METHODS: A retrospective analysis was performed of prospectively collected data from 17 consecutive patients (11 men and 6 women; mean age, 51.7 +/- 16.9 years) treated with RFITT from 2002 to 2003. The postoperative outcome was assessed using active anterior rhinomanometry, "sniffin' stick" test, and endoscopic nasal examination preoperatively and 4 weeks postoperatively. Subjective complaints were assessed with Likert scales. RESULTS: Transnasal airflow increased by 40.3% 4 weeks postoperatively (p = 0.029). Endoscopic appearance of nasal polyps indicated a nonsignificant reduction of 37.1%. Subjective complaints such as impaired nasal ventilation (p = 0.014), nasal discharge (p = 0.0007), postnasal drip (p = 0.0002), and hyposmia (p = 0.048) improved significantly 4 weeks after surgery. CONCLUSION: RFITT is well tolerated as a day case procedure under local anesthesia and might be a procedure for treating recurrence of NP after sinus surgery. It remains unclear at this point whether RFITT for nasal polyps results in a permanent reduction.

Influence of passive humidification on respiratory heat loss in tracheotomized patients.

BACKGROUND: The aim of this study was to evaluate changes in total respiratory heat loss during use of a heat and moisture exchanger (HME) in tracheotomized patients. METHODS: Tracheal humidity and temperature were measured before the application and during use of the HME (plastic foam impregnated with CaCl2), and total respiratory heat loss was calculated. RESULTS: No significant difference was found between the convective heat exchange before and after use of the HME for a 10-minute period. When the HME was placed on the tracheal opening, the evaporative heat exchange and the total respiratory heat loss decreased significantly. CONCLUSIONS: The results indicate that passive airway humidification is effective in tracheotomized patients even after a 10-minute period. However, the positive effect on the energy balance of the tracheal mucosa after prolonged use of the HME remains to be proven.

[Tonsillotomy]. / Die Tonsillotomie.

BACKGROUND: In contrast to tonsillectomy, only intracapsular parts of palatine tonsil are resected in tonsillotomy. This procedure has been revived for treatment of tonsillar hypertrophy causing obstructive sleep apnoea. Tonsillar hypertrophy is the most relevant cause for obstructive sleep apnoea in children often associated with malnutrition due to problems with swallowing. In these patients tonsillotomy is performed, because it is less painful, the children recover more quickly and the results on snoring, difficulty breathing and apnoea are equal in comparison to conventional tonsillectomy. In patients elder than 8 years or suffering from chronic or recurrent tonsillitis, tonsillectomy is the preferable surgical modality, because scarring and persistent inflammation in tonsillar remnants may be urge revision surgery. CONCLUSION: Tonsillotomy is a good alternative for tonsillectomy in children with symptomatic hypertrophy of the palatine tonsils.

[Identification and evaluation of obstructive sleep apnea in children before adenotonsillectomy using evaluative surveys]. / Identifikation und Evaluation der kindlichen Schlafapnoe vor Adenotonsillektomie anhand evaluierter Fragebögen.

Sleep-disordered breathing (SDB) caused by enlarged tonsils and adnoids is common in early childhood. SDB without alterations in alveolar ventilation or sleep architecture is called primary snoring and affects up to 3-12 % of young children. In contrast, SDB with associated apneas or hypopneas affects between 0.7-3 % and is called obstructive sleep apnea syndrome (OSAS). OSAS and primary snoring should be distinguished with polysomnography (PSG). Despite the large number of patients undergoing tonsillectomy and adenoidectomy, the ability to assess patients-based preoperative diagnosis and surgical outcomes polysomnographically is limited. As a consequence it remains difficult which children will or will not benefit from surgical intervention. In the context of OSAS diagnosis, a discriminative survey is desired, one that cold classify differences before surgery as small, moderate or large. A promising office based surrogate measure are disease-specific quality-of-life surveys. Change within patients can be measured using an evaluative survey, administered before and after an intervention.

Selective transduction of malignant glioma by lentiviral vectors pseudotyped with lymphocytic choriomeningitis virus glycoproteins.

Malignant gliomas are the most frequent primary brain tumors and have a dismal prognosis due to their infiltrative growth. Gene therapy using viral vectors represents an attractive alternative to conventional cancer therapies. In a previous study, we established lentiviral vectors pseudotyped with lymphocytic choriomeningitis virus (LCMV) glycoproteins (GPs) and demonstrated transduction of human malignant glioma cells in culture. In the current approach, we compared the transduction efficacy of LCMV-GP- and vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped lentiviral vectors for malignant glioma cells and normal brain cells in vitro and in vivo. LCMV-GP pseudotypes transduced almost exclusively astrocytes, whereas VSV-G pseudotypes infected neurons as well as astrocytes. LCMV-GP pseudotypes showed an efficient transduction of solid glioma parts and specific transduction of infiltrating tumor cells. In contrast, VSV-G-pseudotyped lentiviral vectors transduced only a few tumor cells in solid tumor parts and infected mostly normal brain cells in infiltrating tumor areas. In conclusion, lentiviral vectors pseudotyped with LCMV glycoproteins represent an attractive option for gene therapy of malignant glioma.

Multilevel temperature-controlled radiofrequency therapy of soft palate, base of tongue, and tonsils in adults with obstructive sleep apnea.

OBJECTIVES/HYPOTHESIS: The concept of two-level pharyngeal collapse in patients with obstructive sleep apnea is too simplified. Aggressive multilevel surgeries addressing several airway segments, including skeletal surgery, demonstrate improved success rates. STUDY DESIGN: The study aimed to evaluate the safety and efficacy of multilevel radiofrequency application to soft palate, tonsils, and base of tongue in 16 white patients (mean age, 56.9 +/- 11.1 y; mean body mass index, 27.3 +/- 2.6 kg/m2) with obstructive sleep apnea. There was one dropout. Therapeutic effects after one treatment session were assessed 20.6 +/- 12.6 weeks postoperatively. Treatment outcome measurements were based on Epworth Sleepiness Scale, Likert scales, and polysomnography. METHODS: Every patient received 16 treatment sites with a total dose of 9750 J radiofrequency energy into soft palate, base of tongue, and tonsils. Success was defined as respiratory disturbance index equal to or less than 20 or at least 50% improvement if baseline respiratory disturbance index was less than 20. Statistical analysis was determined with the Spearman rank test. RESULTS: Mean score on Epworth Sleepiness Scale decreased from 11.1 to 8.2 (P =.0001). Of the patients, 53.3% reported improvement of their daytime sleepiness. Snoring was assessed with 10-point Lickert scale, and score decreased from 7.5 to 4.9 (P =.08). Mean respiratory disturbance index decreased from 32.6 +/- 17.4 to 22.0 +/- 15.0 (P =.003). By our definition of success, 5 of 15 patients (33%) have been treated successfully surgically and 4 of 15 (27%) had remarkable improvement after one treatment session; 1 patient (6.6%) demonstrated deterioration. There were two adverse effects, one superficial ulceration of the soft palate and one unilateral tonsillar abscess formation, with an overall complication rate of 13.3% for our patients and 0.41% for all treatment sites (n = 240). CONCLUSION: Moderate to severe obstructive sleep apnea usually requires multilevel pharyngeal surgery. Radiofrequency offers the potential of altering the upper airway on different sites.