Large-scale cultivation of Leishmania infantum promastigotes in stirred bioreactor.

BACKGROUND & OBJECTIVES: Bioreactors are practical tools that are used for economical, time-conserving and large-scale production of biomass from cell cultivation. They provide optimal environmental conditions such as pH and temperature required for obtaining maximum amounts of biomass. However, there is no evidence in the literature on the large-scale cultivation of Leishmania infantum parasites in the bioreactor. Therefore, the present study was undertaken to develop a new approach for obtaining L. infantum biomass by using pH and temperature controllable stirred bioreactor and to compare parasitic growth kinetics with classical method within erlenmeyers. METHODS: In order to obtain parasite biomass, a newly developed pH and temperature controlled stirred bioreactor was used and its efficacy was compared with a graduated classical scale-up method. Growth kinetics of parasites within erlenmeyers and bioreactors were determined by evaluating promastigote numbers using haemocytometer. The graduated scale enlargement of culture was followed by T25 flask, T75 flask, and 1 L erlenmeyer, respectively. RESULTS: Obtained results showed a 10-fold increase in the number of promastigotes within the conventional culture performed in 700 ml medium, while parasite numbers increased approximately 15 times due to initial inoculation amounts in the bioreactor culture performed in the 3.5 l medium. Thus, there was 7.5 times more biomass collection in bioreactor compared to classical method. INTERPRETATION & CONCLUSION: It is postulated that constant culture pH and temperature in the bioreactor extends cultivation time. This could lead to significant increase in parasite numbers. Hence, pH and temperature controllable bioreactors provided acquisition of sufficient amounts of biomass in contrast to classical methods. Therefore, this type of bioreactors may substitute classical culture methods in the production of antigenic molecules for vaccine development.

The impact of frailty on noninvasive mechanical ventilation in elderly medical intensive care unit patients.

AIM: Many factors affecting noninvasive ventilation (NIV) in critically ill patients have been reported in the literature, but there is no study about the effect of frailty. With this study, the frailty prevalence was evaluated with two different frailty scores among the NIV population of a medical intensive care unit (ICU). Besides, the impact of frailty on NIV success and mortality and its association with NIV application problems were evaluated. METHOD: A prospective observational cohort study was performed on patients who were over 50 years of age and assigned to NIV due to hypercapnic respiratory failure. For the assessment of frailty, Clinical Frailty Scale (CFS) and The Edmonton Frailty Scale (EFS) were used and the ones with CFS ≥5 and EFS ≥8 were considered as fragile. The study population was classified and compared according to NIV success, ICU outcome (discharge or exitus) and NIV application problems. RESULTS: A total of 103 patients with the mean age of 73 ± 11 years were included. The incidence of frailty was 41% with CFS ≥5 and 36% with EFS ≥8. The NIV failure occurred in 30 (29%) patients. Among them frailty and SOFA score was higher; Glasgow Coma Scale (GCS) was lower. In multivariate analysis GCS (OR: 1.2, p: 0.042) and frailty with EFS (OR: 2.8, p: 0.027) were identified as independent risk factors of NIV failure. Sixty-five (63%) patients had NIV application problems and frailty was higher among them with both CFS and EFS (p < 0.05). Mortality occurred in 18 (17%) patients; NIV failure and frailty according to CFS were independent risk factors of mortality. CONCLUSION: The frailty is associated with higher NIV application problems, failure and mortality risk in elderly ICU patients. The CFS and EFS frailty scores can be used to predict NIV success and outcomes in ICUs.

Saccharomyces cerevisiae fungemia after probiotic treatment in an intensive care unit patient.

Saccharomyces cerevisiae is a common colonizer of the human gastrointestinal system as a benign organism. Enteral supplementation of this yeast as a probiotic product is effective in the treatment of antibiotic associated diarrhae. In rare occasions it can cause invasive infections. We present two fungemia cases in an intensive care unit following probiotic treatment containing S. boulardii. We are warning the safety of probiotic treatment in critically ill patients.

Overview of dendritic cell-based vaccine development for leishmaniasis.

Leishmaniasis is one of the most serious vector-borne diseases in the world and is distributed over 98 countries. It is estimated that 350 million people are at risk for leishmaniasis. There are three different generation of vaccines that have been developed to provide immunity and protection against leishmaniasis. However, their use has been limited due to undesired side effects. These vaccines have also failed to provide effective and reliable protection and, as such, currently, there is no safe and effective vaccine for leishmaniasis. Dendritic cells (DCs) are a unique population of cells that come from bone marrow and become specialized to take up, process and present antigens to helper T cells in a mechanism similar to macrophages. By considering these significant features, DCs stimulated with different kinds of Leishmania antigens have been used in recent vaccine studies for leishmaniasis with promising results so far. In this review, we aim to review and combine the latest studies about this issue after defining potential problems in vaccine development for leishmaniasis and considering the importance of DCs in the immunopathogenesis of the disease.

[Documentation Time and Effort and Associated Resources for Patients with Primary Breast Cancer from Diagnosis to End of Follow-Up - Results of a Multicentre Validation]. / Dokumentationsaufwand und verbundene Ressourcen bei Patientinnen mit einem primären Mammakarzinom - von der Primärdiagnose bis zur Abschluss der Nachsorge - Ergebnisse der multizentrischen Erhebung.

INTRODUCTION: Tumour documentation is essential for quality assurance of oncological therapies and as a source of reliable information about the in- and outpatient care. The documentation effort and the associated resource consumption were analysed for the example of breast cancer. MATERIAL AND METHODS: The different steps in the care of patients with primary breast cancer in a standardised disease situation were defined from initial diagnosis to the end of the follow-up. After the pilot phase, a multicentre validation (n=7 centres) was performed with the support of the Federal Ministry of Health. The documentation time points were horizontally collected and analysed with regard to amount, duration and personnel expenses. RESULTS: 57% of the documentation costs are caused by the physicians. Regarding the different centres, documentation costs were calculated between € 352.82 and € 1 084.08 per patient from diagnosis to completion of aftercare. Non-certified centres had a reduced documentation effort and thus lower costs. CONCLUSIONS: The results demonstrate the need for a reduction of the documentation effort - particularly for physicians - the most expensive profession in the health system. A quality improvement is expected from the certification with its special requirements. In this context, there is a justified demand for an adequate remuneration of the documentation effort for certified centres. Furthermore, it is necessary to reduce the number of variables for quality assurance and to define them centrally. A comprehensive multi-disciplinary documentation should be achieved. Investments in a single data set and interface enhancements of existing documentation systems should be realised.

Is there any relationship between pulmonary function tests and post-transplant complications of allogeneic hematopoetic stem cell transplantation?

AIM: Pulmonary function tests (PFT) have an important role in the assessment of pulmonary and nonpulmonary complications of hematopoetic stem cell transplantation (HSCT). In this study the relationship between PFTs and DLCOadj values and the complications of HSCT was investigated. The possible role of iron overload in the deterioration of the PFTs after HSCT was also searched. METHODS: One hundred and fifty one patients who had undergone allogeneic HSCT between years 2003 through 2008, and had the records of PFTs prior to and at 1, 3, 6, 9 and 12 months after transplantation were included in the study. Prospectively collected data of these patients were analysed retrospectively. RESULTS: Although no significant difference was identified in other PFT parameters, a significant decrease in DLCOadj was determined after 1st and 3rd months of HSCT. A significant correlation was found between pretransplant DLCOadj value <%70 and sinusoidal obstruction syndrome (SOS) (P=0.001, r=0.323), but in multivariate analysis pretransplant DLCOadj was not an independent predictor of SOS; only total body irradiation (TBI) (OR: 3.673, %95 CI: 0.880-15.804), the day of platelet engraftment (OR=1.093, %95 CI: 1.029-1.161) and serum ferritin (OR=1.001, %95 CI: 1.000-1.001) were significant. Advancing age and serum ferritin levels >600 ng/mL were the independent risk factors for pretransplant DLCOadj <%70 (OR: 0.970, %95 CI: 0.941-0.999 and OR: 2.355, %95 CI: 1.058-5.241 respectively). CONCLUSION: Although a significant correlation exists between pretransplant DLCOadj values and post-transplant SOS development, pretransplant DLCOadj was not an independent predictor of SOS. Increased serum ferritin levels were common both for pretransplant DLCO decrease and post-transplant SOS development. Iron induced endothelial damage may be the common pathophysiologic mechanism causing lung and liver vulnerability, and DLCOadj may be a non-invasive method of demonstrating this vulnerability.

The influence of severe obesity on non-invasive ventilation (NIV) strategies and responses in patients with acute hypercapnic respiratory failure attacks in the ICU.

BACKGROUND: Obesity rates are increasing in the general population and are also prevalent in intensive care units (ICUs). Patients are sometimes admitted to ICUs for hypercapnic respiratory failure or cor pulmonale, but more often, they are admitted for pneumonia, excessive daytime sleepiness, heart failure, chronic obstructive pulmonary disease (COPD), asthma attacks or pulmonary embolism, and hypercapnic respiratory failure is coincidentally noticed during this period. The optimal noninvasive mechanical ventilation strategy is often not used during ICU treatment. The aim of this study was to assess the differences between non-invasive ventilation (NIV) strategies and the outcomes of obese and non-obese patients with acute hypercapnic respiratory failure. METHODS: In this retrospective cohort study, 73 patients who were ventilated with a face mask were studied. Patients were divided into two groups: obese (BMI>35 kg/m2) and non-obese (BMI<35 kg/m2), and the differences between these two groups in necessary pressure, volume, mode, ventilator and time to reduce PaCO2 <50 mmHg were investigated. RESULTS: The mean age of the patients was 66 ± 14 years, and the mean admission APACHE II score was 18 ± 4. Forty-one (56%) of the patients were female. For the obese patients, the reason for ICU admission was more frequently pulmonary edema and less frequently pulmonary infections, which was significantly different (P=0.003 and 0.043, respectively) than the rates for the non-obese patients. While there was no significant difference across the groups between the ventilators, modes and inspiratory pressure levels, obese patients required higher end-expiratory pressure levels and more time to reduce their PaCO2 levels below 50 mmHg than non-obese patients. The lengths of NIV and ICU stay and intubation and the mortality rates were similar in both groups. CONCLUSION: These results suggest that improvement in hypercapnia in obese patients may require higher PEEP levels and longer times than that of non-obese patients during acute hypercapnic respiratory failure attack.

Comparison of the serial surveillance with quantitative and non-quantitative tracheal aspirate in predicting ventilator-associated pneumonia etiology in patients receiving antibiotic therapy.

BACKGROUND: The aim of this study was to investigate the value of serial quantitative (QC) and non-quantitative (NQC) endotracheal aspirate (ETA) surveillance cultures in predicting the causative pathogen of ventilator associated pneumonia (VAP) in patients receiving antibiotic therapy and the factors associated with their predictive value. This was a prospective observational cohort study carried out in the Intensive Care Unit of a tertiary hospital. METHODS: The study enrolled 109 patients receiving mechanical ventilation for at least four days. Tracheal surveillance cultures were obtained routinely thrice weekly. Each sample was processed non-quantitatively and quantitatively (103 and 105 cfu/mL). The sensitivity, specificity and predictive values (true positives plus true negatives) of these cultures for the development of VAP were evaluated, and the causative pathogens were assessed. RESULTS: Sixty-eight VAP episodes were detected during this period. The NQCs and QCs detected the responsible pathogens of VAP in 63% and 28% of the VAP patients, respectively. Surveillance with NQC and QC were negative in 78% and 85% of the patients without VAP, respectively. Compared with the QC-ETAs (47%), NQC-ETAs (65%) were more predictive in all of the 109 patients. The NQC-ETA and QC-ETA predicted the causative pathogens at 3.3+/-2.7 days and 2.5+/-1.7 days prior to the development of VAP episodes, respectively. Both NQC and QC surveillance cultures were less predictive in older patients, and QCs were less predictive in A. baumannii infections. CONCLUSION: These results suggest that surveillance with NQ-ETA is better than the Q-ETA in predicting the development and causative pathogen of VAP in patients who have already been receiving antibiotic therapy.

Risk factors for fungal pulmonary infections in hematopoietic stem cell transplantation recipients: the role of iron overload.

Fungal pulmonary infections (FPIs) are frequent causes of mortality in hematopoietic stem cell transplantation (HSCT) recipients. Determination of the specific risk factors may improve the prognosis. The aim of this study was to evaluate the risk factors of FPIs due to HSCT. Patient history, physical examination, chest X-rays and the consultation records of the pulmonary disease department which were a part of the routine evaluation before and at first, third, sixth, ninth and twelfth months of HSCT were retrieved in 148 adult HSCT recipients. Results of the high-resolution computed tomography, fiber-optic bronchoscopy and the microbiological data were also included. FPI was diagnosed in 22 patients (14.9%). Multivariate analysis showed that increased ferritin levels (>1000 ng/ml; OR: 3.42, 95% CI 1.03-11.42, P=0.045) and the development of sinusoidal obstruction syndrome (SOS; OR: 5.09, 95% CI 1.53-16.90, P=0.008) were significant risk factors for FPIs. The sensitivity and specificity of ferritin >1000 ng/ml for the prediction of FPIs were 67 and 70%, respectively. There was a positive correlation between the increased risk of FPIs and pretransplantation ferritin levels (r=0.413, P<0.001) and increased ferritin levels and SOS (r=0.331, P<0.001). Increased pretransplantation ferritin levels and development of SOS are predictive factors of FPIs during HSCT.

[Advanced ectopic pregnancy--clinical management]. / Fortgeschrittene Extrauteringravidität - klinisches Management anhand einer Fallbeschreibung.

Advanced ectopic pregnancy - clinical management. In case of vaginal bleeding and diffuse abdominal pain you have to include the diagnose of (rare) advanced ectopic pregnancy. Even with ultrasound technique in some of the cases you need further examinations to be sure with your diagnose. Because of the high incidence of maternal morbidity the treatment ist immediate primary laparotomy. In some pregnancies after the 23/24 week there is possibility of a more conservative approach. We describe a case with immediate operative intervention.

[Angiosarcoma of the breast after conservatively operated breast carcinoma--a sequela of adjuvant radiotherapy?]. / Das Angiosarkom der Brust nach konservierend operiertem Mammakarzinom--eine Folge der adjuvanten Nachbestrahlung?

Angiosarcoma represents only 2.4% of soft tissue malignancies. Primary angiosarcoma of the breast is also a rare neoplasm. The incidence of this disease has been reported to be 0.04% of all breast malignancies. The aetiology is unknown. Angiosarcoma associated with chronic lymphoedematous arm of a postmastectomy woman was described by Stewart and Treves. In this paper we describe a 62-year-old female who underwent wide excision, axillary node dissection and adjuvant radiotherapy for primary breast cancer. After 5 years she noticed a purple discolouration of the treated left breast. Biopsy of these areas confirmed an angiosarcoma. No haematogenous dissemination was found. She underwent simple mastectomy without systemic therapy. A chest X-ray 3 months later revealed diffuse bilateral lung metastases. The incidence of breast conserving therapies is growing fast. Until now we have no evidence on the role of irradiation as an aetiological reason for angiosarcoma after breast-conserving surgery and axillary node dissection. Therefore, we have to pay increasing attention to these patients. A patient with history of radiotherapy after breast conserving therapy and skin discolouration must undergo a biopsy of these areas. The only therapy of angiosarcoma is an early mastectomy. The success of chemotherapy is uncertain. Possibly adjuvant chemotherapy with actinomycin D for the group with poorly differentiated tumours appears beneficial. Prospective studies might help to resolve this question.

Vulva reconstruction with a tissue expander.

A patient with exstrophy of the bladder in whom we performed vulva reconstruction using a tissue-expansion technique and local flaps was presented. The ultimate result was in concert with the anatomic characteristics of normal external genitalia with regard to type of skin, pigmentation, hair pattern, quality of circulation, and sensory supply.