[Neutrophils in the tumor microenvironment-foes or friends?] / Neutrophile Granulozyten im Tumormikromilieu ­ Feind oder Freund?

Neutrophils are the most abundant white blood cells in the human circulation and are usually associated with inflammation and with fighting infections. In recent years, the role of these cells during cancer progression has been a matter of increasing interest. Tumor-associated neutrophils (TANs) accumulate in cancer patients and represent an important negative prognostic marker in a broad variety of neoplasms. Accordingly, TANs represent a highly attractive therapeutic target. TAN may exhibit tumor-promoting or -inhibiting functions. Pro-tumor neutrophils support tumor angiogenesis and growth and promote metastatic dissemination of tumors via establishment of the premetastatic niche. Studies in animal models have already shown that the depletion of TANs or the inhibition of their migration bears therapeutic potential. Multiple pathways and mediators that induce pro-tumoral functions in neutrophils have been identified. In this review, we provide an up-to-date overview of the pro- and anti-tumor properties of neutrophils as well as the environmental cues that regulate these distinct functions. We also report on our own work that comprehensively investigated the role of neutrophils in head and neck cancer.

Metabolite profiles evaluated, according to sex, do not predict resting energy expenditure and lean body mass in healthy non-obese subjects.

PURPOSE: Differences in resting energy expenditure (REE) between men and women mainly result from sex-related differences in lean body mass (LBM). So far, a little is known about whether REE and LBM are reflected by a distinct human metabolite profile. Therefore, we aimed to identify plasma and urine metabolite patterns that are associated with REE and LBM of healthy subjects. METHODS: We investigated 301 healthy male and female subjects (18-80 years) under standardized conditions in the cross-sectional KarMeN (Karlsruhe Metabolomics and Nutrition) study. REE was determined by indirect calorimetry and LBM by dual X-ray absorptiometry. Fasting blood and 24 h urine samples were analyzed by targeted and non-targeted metabolomics methods using GC × GC-MS, GC-MS, LC-MS, and NMR. Data were evaluated by predictive modeling of combined data using different machine learning algorithms, namely SVM, glmnet, and PLS. RESULTS: When evaluating data of men and women combined, we were able to predict REE and LBM with high accuracy (> 90%). This, however, was a clear effect of sex, which is supported by the high degree of overlap in identified important metabolites for LBM, REE, and sex, respectively. The applied machine learning algorithms did not reveal a metabolite pattern predictive of REE or LBM, when analyzing data for men and women, separately. CONCLUSIONS: We could not identify a sex independent predictive metabolite pattern for REE or LBM. REE and LBM have no impact on plasma and urine metabolite profiles in the KarMeN Study participants. Studies applying metabolomics in healthy humans need to consider sex specific data evaluation.

High frequency of herpesvirus-specific clonotypes in the human T cell repertoire can remain stable over decades with minimal turnover.

High-throughput T cell receptor sequencing on sequentially banked blood samples from healthy individuals has shown that high-frequency clonotypes can remain relatively stable for up to 18 years, with minimal inflation, deflation, or turnover. These populations included T cell expansions specific for Epstein-Barr virus. Thus, in spite of exposure to a barrage of microorganisms over the course of life, the dominant clonotypes in the mature peripheral T cell repertoire can alter surprisingly little.

Association of an RNA kissing complex analyzed using 2-aminopurine fluorescence.

The fluorescent probe, 2-aminopurine-2'-O-methyl riboside (2-AP) has been selectively incorporated at adenosine positions in stem-loops (so called R1inv and R2inv), derived from the ColE1 plasmid encoded RNA I and RNA II transcripts, that interact to form stable loop-loop kissing complexes and bind the RNA one modulator (Rom) protein, such that fluorescence-detected stopped-flow and equilibrium methods could be used to study the detailed mechanism of this RNA-RNA interaction. Formation of loop-loop kissing complexes between R1inv and R2inv hairpins, substituted with 2-AP at positions in the complementary loops, results in a 5-10-fold fluorescence emission decrease (F(max) = 370 nm), which provides a sensitive measure for the binding reaction. The 2-AP substituted complexes are found to have equilibrium binding properties (average K(D) = 2.6 +/- 1.7 nM) and affinity for Rom (average K(D) = 60 +/- 24 nM) that are similar to complexes formed with equivalent unlabeled hairpins. Using stopped-flow experiments, it was found that the 2-AP probes experienced at least three different microenvironments during association of the RNA complex, thus suggesting a kinetic intermediate in the kissing pathway. In contrast, dissociation of the complex was found to fit a single exponential decay (average k(off) = 8.9 x 10(-5) s(-1)). Consistent with these observations, a two-step mechanism for RNA loop-loop complex association is proposed in which the complementary loops of R1inv and R2inv first base pair to form the loop-loop helix (average k(1) = 0.13 microM(-1)s(-1)) in the initial encounter reaction, and subsequently isomerize to the final tertiary fold in a second slower step (average k(2) = 0.09 s(-1)), where the helical stacking around the junctions is optimized.

High-throughput tissue microarray analysis of 3p25 (RAF1) and 8p12 (FGFR1) copy number alterations in urinary bladder cancer.

Studies by comparative genomic hybridization revealed that the chromosomal regions 3p25 and 8p11-p12 are recurrently amplified in bladder cancer. To investigate the prevalence of DNA copy number alterations in these chromosomal regions and study their clinical significance, we used probes for the RAF1 (3p25) and FGFR1 (8p12) genes for fluorescence in situ hybridization. A tissue microarray containing 2317 tumors was analyzed. The analysis revealed RAF1 amplification in 4.0% and FGFR1 amplification in 3.4% of interpretable tumors. In addition, deletions were found at the 3p25 locus in 2.2% and at the 8p11-12 locus in 9.9% of interpretable tumors. Both amplifications and deletions of RAF1 and FGFR1 were significantly associated with high tumor grade (P < 0.0001), advanced stage (P < 0.0001), and poor survival (P < 0.05) if tumors of all of the stages where analyzed together. RAF1 amplifications were associated with subsequent tumor progression in pT1 carcinomas (P < 0.05). The marked differences in the frequency of all of the analyzed changes between pTa grade 1/grade 2 and pT1-4 carcinomas support the concept of these tumor groups representing different tumor entities.

Ultrafast energy transfer and structural dynamics in DNA.

Ultrafast structural dynamics concomitant to excitation energy transfer in DNA has been studied using a pair of pyrene-labeled DNA bases. The temporal evolution of the femtosecond pump-probe spectra reveals the existence of two electronic coupling pathways, through-base stack and through-space, which lead to excitation energy transfer and excimer formation even when the labeled DNA bases are separated by one AT base pair. The electronic coupling which mediates through-base stack energy transfer is so strong that a new absorption band arises in the excited-state absorption spectrum within 300 fs. From the analysis of time-dependent spectral shifts due to through-space excimer formation, the local structural dynamics and flexibility of DNA are characterized on the picosecond and nanosecond time scale.

Polysomies but not Y chromosome losses have prognostic significance in pTa/pT1 urinary bladder cancer.

A disturbed cellular DNA content is of potential diagnostic and prognostic relevance in urinary bladder cancer. To evaluate the prognostic significance of individual chromosomal aberrations in superficial bladder cancer, specimens of 105 tumors (67 pTa, 38 pT1) were examined by fluorescence in situ hybridization (FISH). FISH allows quantitation of chromosomes on a cell by cell level. Centromere probes for the chromosomes Y, 1, and 17 were used. There was a strong association between polysomies of the chromosomes 1 (found in 46% of tumors) and 17 (40% of tumors, P < .0001). Polysomies (1 and 17) were significantly more frequent in pT1 than in pTa tumors (P < .0001 each). In pTa tumors, polysomies of both chromosomes were linked to a high risk of recurrences; polysomy 17 was associated with an increased risk of progression (P < .05 each). There was no significant association between polysomies and an unfavorable prognosis in pT1 carcinomas. Previous studies had suggested a prognostic role of Y losses in bladder cancer. However, Y losses were not linked to recurrences or tumor progression in pTa or pT1 tumors of 67 male patients. These data show that marked genetic differences exist between pTa and pT1 carcinomas. They also indicate that polysomies of different chromosomes may have prognostic relevance in pTa urinary bladder cancer.

Construction of improved plasmid vectors for promoter characterization in Pseudomonas aeruginosa and other gram-negative bacteria.

We report the construction of two broad host range promoter-probe plasmid vectors for rapid analysis of promoters in Gram-negative bacteria. The new vectors, pME4507 and pME4510, carry carbenicillin and gentamycin resistance genes, respectively, and are small sized (4 kb) with a flexible multiple cloning site to facilitate directional cloning of putative promoter elements. The vectors allow rapid plate-based screening for promoter activities, using beta-galactosidase as the reporter enzyme. In the absence of an inserted promoter fragment, they display very low background activity, making them a useful tool for analysis of low expression level promoters.

Regional cerebral blood flow in man at rest and during exercise.

Regional cerebral blood flow (rCBF) of the left hemisphere was measured in 12 healthy young men at rest and during physical work on a bicycle ergometer in the supine position at work-load levels of 25 W or 100 W using the intravenous 133Xe method. Regional mean cerebral blood flow, regional gray-matter flow, and relative gray-matter weight was determined for six regions of interest. Arterial blood pressure, pulse frequency and expiratory CO2 concentration were recorded. Cerebral blood flow in all regions was significantly (P less than 0.001) higher during exercise than at rest. The increase in the 100 W group (24.7%) was significantly (P less than 0.05) greater than in the 25 W group (13.5%), but resting blood flow levels and alveolar CO2 concentrations were also different in both groups. Mean arterial blood pressure, pulse frequency and alveolar CO2 concentrations, but not arterial pCO2, were significantly higher during exercise and there was a faster washout of whole-body xenon. The CBF increase was interpreted as a combined effect of elevated systemic blood pressure and functionally activated brain metabolism. There was no evidence of impaired cerebral autoregulation.

Clinical value of a mucin-like carcinoma-associated antigen (MCA) in patients with carcinomas of the kidney and the urinary collecting system.

A new tumor marker, mucin-like carcinoma-associated antigen (MCA), was evaluated in thirty-four patients with renal cell carcinomas and forty patients with carcinomas of the urinary collecting system. In the first group, specificity was 95%, sensitivity 65%, the predictive value for positive diagnosis was 94%, and for negative diagnosis 72%. In the second group specificity was 83%, sensitivity 71%, predictive value for positive diagnosis was 63%, and for negative diagnosis 88%. Receiver operating characteristic curves indicated that the maximum amount of information was greater in patients of the first group than the second (0.379 vs. 0.332 bits). In the light of these data, the MCA test can be recommended for assessing metastatic spread in patients particularly with renal cell carcinomas.

Preoperative predictors of recurrent atrial fibrillation late after successful mitral valve reconstruction.

OBJECTIVE: Late outcome after mitral valve repair was examined to define preoperative predictors of recurrent atrial fibrillation late after successful mitral valve reconstruction. METHODS: One hundred and eighty-nine patients, 112 with preoperative sinus rhythm and 72 with preoperative chronic or intermittent atrial fibrillation, were followed for 12.2 +/- 10 years after valve repair. Clinic, hemodynamic end echocardiographic data were entered into Cox-regression and Kaplan-Meyer analysis to assess predictors for recurrent atrial fibrillation late after successful mitral valve repair. RESULTS: Univariate and multivariate predictors for recurrent atrial fibrillation late after successful mitral valve reconstruction were preoperative atrial fibrillation (P = 0.0001), preoperative antiarrhythmic drug treatment (P = 0.005), heart rate (P = 0.01), left ventricular ejection fraction (P = 0.01) and increased left ventricular posterior wall thickness (P = 0.05). Patients > 57.5 years with a mean pulmonary artery pressure > or =23 mm Hg and a history of preoperative antiarrhythmic drug treatment had an odds ratio of 53.33 (95% confidence limits 6.12-464.54) for atrial fibrillation late after successful mitral valve repair. CONCLUSION: Older patients with a history of atrial fibrillation, antiarrhythmic treatment or an elevated pulmonary artery pressure may present atrial fibrillation late after successful mitral valve repair. They could be considered for combined mitral valve reconstruction and surgery for atrial fibrillation even though sinus rhythm is present preoperatively.

Influence of pneumoperitoneum and patient positioning on respiratory system compliance.

STUDY OBJECTIVE: To investigate the influence of pneumoperitoneum (PP) and posture on respiratory compliance and ventilation pressures. DESIGN: Prospective, single blind trial. PATIENTS: 10 female ASA physical status I and II patients scheduled for elective gynecologic laparoscopy. SETTING: University medical center. INTERVENTIONS: Anesthesia was performed as total IV anesthesia (TIVA) with propofol, alfentanil, and atracurium. After induction of anesthesia and orotracheal intubation, the lungs were ventilated to maintain partial pressure of CO(2) (P(ET)CO(2)) of 30 +/- 3 mmHg. Ventilation was kept constant. As gas mixture oxygen and air 1:1 was used without positive end-expiratory pressure (PEEP). MEASUREMENTS: Measurements were taken before and after creation of pneumoperitoneum with an intraabdominal pressure (IAP) of 10 mmHg, of 15 mmHg in 20 degrees head-down tilt, then in 20 degrees head-up tilt, and after deflation of PP. We determined peak inspiratory pressure (PIP), mean airway pressure (mPaw), P(ET)CO(2), expiratory minute volume (V(E)), heart rate (HR), and systolic (SBP), diastolic (DBP), and mean arterial pressure (MAP). Respiratory system compliance (C(eff rs)) was calculated as quotient of tidal volume (V(T)) and PIP. MAIN RESULTS: After creation of PP (IAP 10 mmHg), there was a significant increase of median PIP (3 cmH(2)O), mPaw (1 cm H(2)O) and arterial pressure (BP), (MAP by 7 mmHg), C(eff rs) decreased by 6 mL. cm H(2)O(-1). Increase of IAP to 15 mmHg led to a further increase of PIP (2 cm H(2)O) and mPaw (1 cm H(2)O), and a further decrease of C(eff rs) by 5 mL cm H(2)O(-1); BP decreased (MAP by 5.5 mmHg). Head-up or head down positions showed no significant hemodynamic or pulmonary changes. P(ET)CO(2)increased from 29.5 to 36 mmHg at an IAP of 15 mmHg, but then no further changes were noticed. Five minutes after deflation of pneumoperitoneum all values returned to baseline levels. CONCLUSIONS: Creation of PP at an IAP of 15 mmHg reduced respiratory system compliance, and increased peak inspiratory and mean airway pressures, which quickly returned to normal values after deflation. Head-down or head-up position did not further alter those parameters.

Influence of pneumoperitoneum and patient positioning on preload and splanchnic blood volume in laparoscopic surgery of the lower abdomen.

STUDY OBJECTIVE: To determine the hemodynamic effects of pneumoperitoneum and patient positioning during laparoscopic surgery of the lower abdomen. DESIGN: Prospective study. SETTING: University-affiliated medical center. PATIENTS: 10 ASA physical I and II female patients scheduled for laparoscopic surgery of the lower abdomen. INTERVENTIONS: Patients were anesthetized with propofol and an alfentanil infusion, then intubated, and normoventilated. MEASUREMENTS: After intubation, a transesophageal multiplane probe for measurements of right (RVESA) and left (LVESA) ventricular end-systolic and end-diastolic areas (RVEDA and LVEDA) and ejection fraction area (RVEFa, LVEFa) was introduced; heart rate (HR) and noninvasive blood pressure (BP) were recorded every minute. Ventilation was not changed during the measurements. A transvaginal ultrasound probe was inserted to measure the diameter of the common iliac vein. Measurements were performed 15 minutes after induction of anesthesia and while patients were in the supine position (P 0), 10 minutes after CO(2) insufflation to 10 mmHg IA pressure (P 10), 10 minutes after a further increase to 15 mmHg (P 15), 10 minutes after 20 degrees Trendelenburg (P 15 T), and 20 degrees reverse Trendelenburg positions (P 15 RT). Data are shown as medians, 25th to 75th percentiles, and comparisons between P 0, P 10, P 15, and P15 T were made with the Friedman test, followed by Wilcoxon test, when significant. Data at P 15 T, P 15 RT, and P 15 were compared using the Wilcoxon test, with a p-value < 0.05 regarded as significant. MAIN RESULTS: Pneumoperitoneum at 10 mmHg abdominal pressure caused a significant increase of LVESA by 78% (RVESA: 61%) and LVEDA by 48.5% (RVEDA: 45%). The diameter of the common iliac vein was decreased by 6%. A further increase of abdominal pressure to 15 mmHg led to an additional increase of 20% (LVESA) and 17% (LVEDA). Mean arterial pressure increased by a significant 7% at P 10, decreasing subsequently by 5% at P 15. The Trendelenburg position did not alter any hemodynamic findings. Reverse Trendelenburg position, however, caused a significant LVEDA-and RVEDA-decrease by 18% and 27%, respectively, and an increase in the diameter of the common iliac vein by 22%. The LVEFa and RVEFa decreased significantly after abdominal CO(2) insufflation by 18% each (P 10) without further change. CONCLUSIONS: The lithotomy position and subsequent pneumoperitoneum increased preload, probably as a result of blood shifting from the abdomen to the thorax by compression of splanchnic vessels caused by the pneumoperitoneum. Careful fluid management, maintaining low abdominal pressure, and use of the reverse Trendelenburg position are favored to prevent adverse hemodynamic effects in laparoscopic surgery.

[Familial adult cystic kidneys and malignant teratomas after kidney transplantation with cyclosporin therapy]. / Familiäre adulte Zystennieren und maligne Teratome nach Nierentransplantation unter Ciclosporintherapie.

We report on two brothers with adult polycystic kidney disease, malignant teratomas and other genital malformations. Because of the unusual accumulation of malformations of embryologically related organs, we postulate a connection between malformations of the kidneys and the genital tract, on the one hand, and teratomas on the other. No genetic coherence is known so far. It is unlikely that immunosuppression with cyclosporin after transplantation had caused these tumours.