Effect of hypoxia on tissue factor pathway inhibitor expression in breast cancer.

UNLABELLED: ESSENTIALS: A hypoxic microenvironment is a common feature of tumors that may influence activation of coagulation. MCF-7 and SK-BR-3 breast cancer cells and breast cancer tissue samples were used. The results showed transcriptional repression of tissue factor pathway inhibitor expression in hypoxia. Hypoxia-inducible factor 1α may be a target for the therapy of cancer-related coagulation and thrombosis. BACKGROUND: Activation of coagulation is a common finding in patients with cancer, and is associated with an increased risk of venous thrombosis. As a hypoxic microenvironment is a common feature of solid tumors, we investigated the role of hypoxia in the regulation of tissue factor (TF) pathway inhibitor (TFPI) expression in breast cancer. OBJECTIVES: To explore the transcriptional regulation of TFPI by hypoxia-inducible factor (HIF)-1α in breast cancer cells and their correlation in breast cancer tissues. METHODS AND RESULTS: MCF-7 and SK-BR-3 breast cancer cells were cultured in 1% oxygen or treated with cobalt chloride (CoCl2 ) to mimic hypoxia. Time-dependent and dose-dependent downregulation of TFPI mRNA (quantitative RT-PCR) and of free TFPI protein (ELISA) were observed in hypoxia. Western blotting showed parallel increases in the levels of HIF-1α protein and TF. HIF-1α inhibitor abolished or attenuated the hypoxia-induced downregulation of TFPI. Luciferase reporter assay showed that both hypoxia and HIF-1α overexpression caused strong repression of TFPI promoter activity. Subsequent chromatin immunoprecipitation and mutagenesis analysis demonstrated a functional hypoxia response element within the TFPI promoter, located at -1065 to -1060 relative to the transcriptional start point. In breast cancer tissue samples, gene expression analyses showed a positive correlation between the mRNA expression of TFPI and that of HIF-1α. CONCLUSIONS: This study demonstrates that HIF-1α is involved in the transcriptional regulation of the TFPI gene, and suggests that a hypoxic microenvironment inside a breast tumor may induce a procoagulant state in breast cancer patients.

Hyperfibrinolysis is common in out-of-hospital cardiac arrest: results from a prospective observational thromboelastometry study.

BACKGROUND: Cardiocirculatory arrest (CCA) activates procoagulant pathways. It has also been reported to inhibit fibrinolysis, resulting in fibrin deposition and further impairment of blood flow. Until now, no studies have used whole-blood viscoelastic tests to characterize coagulation and the impact of fibrinolysis in out-of-hospital cardiac arrest (OHCA). METHODS: Patient with established OHCA who underwent cardiopulmonary resuscitation (CPR) were enrolled. Blood samples were obtained immediately after placement of an intravenous line at the scene, for full blood cell count, standard coagulation tests and rotational thromboelastometric (ROTEM(®)) analyses. Patients with return of spontaneous circulation (ROSC) were compared to non-ROSC patients. RESULTS: Fifty-three patients (median age 67 years, interquartile range: 56-73 years) were included in the study. ROSC was established in 25 patients. Prothrombin time index (PTI) was significantly lower and activated partial thromboplastin time (aPTT) was significantly prolonged in non-ROSC patients compared to ROSC patients. Clotting time (CT) in the extrinsically activated ROTEM test (EXTEM) was significantly longer in non-ROSC versus ROSC patients. For the remaining EXTEM parameters, there were no significant differences between ROSC and non-ROSC patients. Hyperfibrinolysis (maximum lysis>15% according to ROTEM test results) was observed in 19 patients (35.8%). There was no difference between ROSC and non-ROSC patients in the incidence of hyperfibrinolysis. CONCLUSIONS: PTI, aPTT and EXTEM CT revealed significant differences between ROSC and non-ROSC patients. Hyperfibrinolysis according to ROTEM test results was much more common than previously assumed. Routine use of fibrinolytic therapy in all patients with prolonged CPR cannot therefore be recommended.

Implantation metastasis of prostate cancer in a suprapubic cystostomy tube needle tract.

A 64-year-old male patient with a serum PSA value of 51.91 ng ml(-1) and an magnetic resonance imaging scan suggestive of prostate cancer was admitted with bilateral hydronephrosis and anemia. A suprapubic cystostomy had been inserted 6 months earlier, owing to voiding difficulties. The patient was found to have huge multiple bladder stones and a massively thickened bladder wall. A sectio alta with bladder stone removal and bilateral ureteral stenting was performed. Histopathological examination revealed the growth of prostate cancer cells in the cystostomy tube needle tract.

The multi-herbal drug STW 5 (Iberogast) has prosecretory action in the human intestine.

There is growing evidence that STW 5 (Iberogast), fixed combination of hydroethanolic herbal extracts), besides being effective in functional dyspepsia, also improves symptoms in irritable bowel syndrome (IBS). Clinical data indicate that modulation of mucosal secretion is a promising approach to treat intestinal disorders associated with IBS. We therefore explored the effect of STW 5 on secretion in the human intestine and the mechanisms by which it acts. The Ussing chamber technique was used to measure mucosal secretion in human intestinal mucosa/submucosa preparations and in human epithelial cell line T84. In addition, we recorded STW 5 effects on human enteric neurons with voltage sensitive dye imaging. In human tissue and T84 cells STW 5 induced a dose-dependent increase in ion secretion that was significantly reduced by the Na-K-Cl cotransporter blocker bumetanide, the adenylate cyclase inhibitor MDL-12 330, the non-specific and selective cystic fibrosis transmembrane conductance regulator (CFTR) inhibitors glibenclamide and CFTR(inh)-172, respectively, and the blocker of calcium dependent Cl(-) channels (ClCa) SITS (4-acetamido-4-isothiocyanatostilbene-2,2-disulphonic acid). It was unaffected by amiloride, a blocker of epithelial Na(+) channels. In human tissue, the nerve blocker tetrodotoxin significantly suppressed the STW 5 response. STW 5 evoked an increased spike discharge in 51% of human submucous neurons. Results suggest that STW 5 is a secretogogue in the human intestine by direct epithelial actions and through activation of enteric neurons. The prosecretory effect is due to increased epithelial Cl(-) fluxes via CFTR and Ca-dependent ClCa channels. STW 5 may be a novel option to treat secretory disorders associated with IBS and constipation.

Optically induced hybridization of a quantum dot state with a filled continuum.

We present an optical signature of a hybridization between a localized quantum dot state and a filled continuum. Radiative recombination of the negatively charged trion in a single quantum dot leaves behind a single electron. We show that in two regions of vertical electric field, the electron hybridizes with a continuum through a tunneling interaction. The hybridization manifests itself through an unusual voltage dependence of the emission energy and a non-Lorentzian line shape, features which we reproduce with a theory based on the Anderson Hamiltonian.

True adenomas of the cardia: a case series of 3 patients.

BACKGROUND/AIM: True adenomas of the cardia appear to be extremely rare lesions. There are no data on the natural history and histopathological background of these lesions. We report 3 patients with true adenomas of the cardia. METHODS AND RESULTS: Three patients with polypoid masses at the cardia below the Z-line were submitted to a tertiary referral center for further diagnosis and therapy. In 2 of the 3 cases Barrett's esophagus with low-grade intraepithelial neoplasia was assumed on the basis of histopathological examination of biopsy specimens taken from the surface of the lesions. Polypectomy was performed in all 3 cases. In 2 of the 3 cases the final histopathological diagnosis of low-grade adenoma of the cardia could only be established after complete removal of the polypoid masses. CONCLUSIONS: Adenomas of the cardia can be mistaken for dysplasia arising from Barrett's esophagus, if the diagnosis is based on endoscopic biopsies only. It is, therefore, reasonable to completely remove any suspicious lesions by endoscopy not only for therapeutic but also for diagnostic reasons.

The nonlinear Fano effect.

The Fano effect is ubiquitous in the spectroscopy of, for instance, atoms, bulk solids and semiconductor heterostructures. It arises when quantum interference takes place between two competing optical pathways, one connecting the energy ground state and an excited discrete state, the other connecting the ground state with a continuum of energy states. The nature of the interference changes rapidly as a function of energy, giving rise to characteristically asymmetric lineshapes. The Fano effect is particularly important in the interpretation of electronic transport and optical spectra in semiconductors. Whereas Fano's original theory applies to the linear regime at low power, at higher power a laser field strongly admixes the states and the physics becomes rich, leading, for example, to a remarkable interplay of coherent nonlinear transitions. Despite the general importance of Fano physics, this nonlinear regime has received very little attention experimentally, presumably because the classic autoionization processes, the original test-bed of Fano's ideas, occur in an inconvenient spectral region, the deep ultraviolet. Here we report experiments that access the nonlinear Fano regime by using semiconductor quantum dots, which allow both the continuum states to be engineered and the energies to be rescaled to the near infrared. We measure the absorption cross-section of a single quantum dot and discover clear Fano resonances that we can tune with the device design or even in situ with a voltage bias. In parallel, we develop a nonlinear theory applicable to solid-state systems with fast relaxation of carriers. In the nonlinear regime, the visibility of the Fano quantum interferences increases dramatically, affording a sensitive probe of continuum coupling. This could be a unique method to detect weak couplings of a two-level quantum system (qubits), which should ideally be decoupled from all other states.

Comparison of laser and mercury-arc lamp for the detection of body fluids on different substrates.

The performance of two detection techniques for body fluids, the Spectra-Physics(R) Revealtrade mark portable forensic laser system and the mercury-arc lamp Lumatec Superlite 400, was evaluated with various biological stains on different substrates. Serial dilutions of neat, 1/10, 1/100 and 1/1,000 using fluid semen, saliva, urine and blood were applied on glazed tiles, glass, PVC, wood, metal, stone, formica, carpet and cotton. Apart from the fact that blood traces were not detectable with the laser, both light sources showed comparable results regarding their detection capability. Clear advantages of the Lumatec Superlite 400, however, are its lower size, weight, purchase costs and the possibility to operate this light source by battery.

Severity and duration of mental deficiency symptoms after intravenous administration of propofol.

The severity and duration of cognitive performance capacity deficits after intravenous administration of propofol were determined using the validated psychological test procedure syndrome short test (SKT), a simple reaction test and original driving licence exam questions. The test battery was performed before, immediately after, as well as 1 and 2 h after propofol administration in 23 persons. Immediately after propofol anaesthesia, six individuals had a slight performance loss, and four subjects showed mild deficits, consistent with medium organic neuropsychologic disorder or dementia. The status of the subjects rapidly changed for the better, and 2 h after propofol anaesthesia, only one person (4%) showed slight deficits of memory and attention. Therefore, it is suggested that patients refrain from any participation in road traffic for at least 2 h after propofol anaesthesia. Driving a car should not be admitted until an interval of 6 h has elapsed.

Hypoxic changes in Purkinje cells of the human cerebellum.

The significance of both Purkinje cell numbers and various neuronal changes for the diagnosis and timing of hypoxic-induced brain lesions was investigated in tissue samples from the cerebellar cortex of 52 individuals with a history of acute or prolonged cerebral hypoxia/ischemia before death. Furthermore, the area of the Purkinje cell somata (PC size) was measured using an automatic image processing and analysis system (LEICA QWin). Significantly reduced numbers of Purkinje cells (<6 cells/unit length of 1 mm) and a decreased portion (<50%) of intact Purkinje cells could be detected in individuals with a period of resuscitation of at least 2 h after acute circulatory arrest. Average cell numbers of less than 4 cells/unit were found in individuals who suffered from diffuse brain swelling and were ventilated for at least 3 days, as well as in individuals who died of brain death. Moreover, the Purkinje cells in these cases exhibited shrunken somata compared to the controls. Specimens that were stored at room temperature up to 30 h after removal at autopsy showed no significant autolytic changes of the Purkinje cells. After 46 h, however, reduced Purkinje cell numbers and shrunken cell bodies were found.

Value of morphological parameters for grading of brain swelling.

This study investigated the value of both gross features and histological findings for grading of brain swelling. For this purpose, the grooving of the temporal gyri (unci) as well as the extension of the cones at the basal part of the cerebellum were measured in 42 brains obtained at autopsy. Furthermore, the distension of perivascular spaces in tissue samples from seven different regions of the brains was evaluated histologically, assisted by an automatic image processing and analysis system. In each individual, the normal range of brain weight was calculated on the basis of the body height, using the formulae by Röthig and Schaarschmidt. The difference between this calculated (normal) value and the brain weight evaluated at autopsy was considered as a reliable criterion for the grade of brain swelling. There was no statistical evidence of a positive correlation between the various parameters. Hence, it can be concluded that both gross section and histological findings are of minimal significance for grading of brain swelling.

Human mast cell mediator cocktail excites neurons in human and guinea-pig enteric nervous system.

Neuroimmune interactions are an integral part of gut physiology and involved in the pathogenesis of inflammatory and functional bowel disorders. Mast cells and their mediators are important conveyors in the communication from the innate enteric immune system to the enteric nervous system (ENS). However, it is not known whether a mediator cocktail released from activated human mast cells affects neural activity in the ENS. We used the Multi-Site Optical Recording Technique to image single cell activity in guinea-pig and human ENS after application of a mast cell mediator cocktail (MCMC) that was released from isolated human intestinal mucosa mast cells stimulated by IgE-receptor cross-linking. Local application of MCMC onto individual ganglia evoked an excitatory response consisting of action potential discharge. This excitatory response occurred in 31%, 38% or 11% neurons of guinea-pig submucous plexus, human submucous plexus, or guinea-pig myenteric plexus, respectively. Compound action potentials from nerve fibres or fast excitatory synaptic inputs were not affected by MCMC. This study demonstrates immunoneural signalling in the human gut and revealed for the first time that an MCMC released from stimulated human intestinal mast cells induces excitatory actions in the human and guinea-pig ENS.

A predictive model for endometriosis.

BACKGROUND: Aromatase is the key enzyme in the process of estrogen biosynthesis from the precursor androgen. Recently, aromatase has been found to be aberrantly expressed in eutopic endometrium of patients suffering from endometriosis. This finding has prompted speculation about the contribution of this enzyme to the prediction of this disease. METHODS: We prospectively aimed to evaluate whether endometrial biopsy, prior to laparoscopy in symptomatic women to screen for the presence of aromatase by real-time RT-PCR and immunohistochemistry, combined with select patients' characteristics, is of value to predict endometriosis. RESULTS: Of 48 consecutive symptomatic and eligible patients, 25 (52.1%) exhibited endometriosis and 23 (47.9%) were disease-free. A multiple logistic regression model revealed that 95.5% of patients whose eutopic endometrium was found to be positive for aromatase mRNA as well as immunohistochemically detected protein and who were additionally suffering from moderate to severe dysmenorrhoea (visual analogue scale score >4/10) exhibited endometriosis at laparoscopy. CONCLUSIONS: These findings provide direct evidence that screening for eutopic endometrial aromatase in combination with clinical data could be of discriminative value in the prediction of disease.

Cardiac involvement in Emery-Dreifuss muscular dystrophy.

Emery-Dreifuss muscular dystrophy (EDMD) is a common form of muscular dystrophy frequently involving cardiac muscle, thus leading to dilated cardiomyopathy. Clinical outcome and prognosis is frequently determined by the involvement of the cardiac conduction system causing symptomatic bradyarrhythmias, as well as tachyarrhythmias and, if untreated, frequent sudden cardiac death. Typical features of the cardiac involvement of EDMD are presented, caused by a novel missense mutation in the splice receptor sequence of intron 6 of the LMNA gene on chromosome 1, encoding for the lamin A/C gene, consistent with the autosomal dominant form of EDMD.

[Endoscopic, pathologic-anatomic and histologic findings in the bovine teat. 2. Changes in the area of Fürstenberg's rosette]. / Endoskopische, pathologisch-anatomische und histologische Befunde an der Rinderzitze. 2. Veränderungen im Bereich der Fürstenbergschen Rosette.

In the present study endoscopically diagnosed changes of the Fürstenberg's rosette were examined histologically. For this purpose the teats of 200 slaughtered cows were inspected. In 19 cases proliferations in the region of the Fürstenberg's rosette were found. Additionally, tissue samples were taken from the Fürstenberg's rosette of 26 cows which showed milk flow disorders due to stenoses of this part of the teat. Teats of slaughtered cows as well as biopsy specimens were fixed in 5% formaldehyde and embedded in paraffin and in plastic. All specimens were stained according to H.E., Giemsa and Turnbull, respectively. Histologically, the proliferations of the Fürstenberg's rosette consisted of fibrovascular tissue (granulation tissue) covered by keratinized squamous epithelium (teat canal epithelium) in eleven cases. In the other cases double-layered cuboid epithelium (cisternal epithelium) was found additionally. The formation of granulation tissue and deposition of blood pigment was observed exclusively in areas covered by stratified epithelium. Therefore a traumatic lesion of the teat canal was supposed to be the cause of the histological findings. In contrast to several reports in the literature, however, no indications of an inversion of the teat canal epithelium were found. On the basis of these findings, the hitherto assumed idea of pathogenesis of proliferations of the Fürstenberg's rosette was modified and a new pathogenesis pattern was developed.

[Endoscopic, pathologic-anatomic and histologic findings in the bovine teat. 1. Changes in the area of the teat cistern in slaughtered cows]. / Endoskopische, pathologisch-anatomische und histologische Befunde an der Rinderzitze. 1. Veränderungen im Bereich der Zitzenzisterne bei Schlachtkühen.

In the present study 795 teats of slaughtered cows were examined endoscopically. Teats which showed lesions were examined histologically. Abnormalities were detected endoscopically in 37 teats. In five cases red discolorations were observed. Histologically these alterations proved to be fresh bleedings within the fibromuscular layer of the teat cistern. Thirteen teats showed yellowish discolorations of the cistern surface. Three of these cases could not be cleared up histologically. In the other cases moderate or severe blood pigment deposition (n = 10) or severe fibrosis (n = 3) of the fibromuscular layer were found. Neither fresh bleeding nor severe blood pigment deposition was found in teats with normal coloured epithelium of the cistern. Cistern surface was smooth in 18 cases, whereas in 19 cases nodules were found. Histologically all nodules consisted of fibrovascular tissue ("granulation tissue") covered with cisternal epithelium. Detailed histological examinations, however, revealed considerable morphological variations regarding the overlying epithelium, the extent and kind of cellular infiltration, the extent of angioplasia and fibroplasia as well as fresh bleedings or deposition of blood pigment in the underlying fibrovascular tissue in some cases. It was concluded, that teat endoscopy enables a differentiation between fresh and old bleedings according to the coloration of the cisternal surface. Nodular proliferations of the cistern surface generally consists of fibrovascular ("granulation") tissue. Further histological details, however, and thus possible information about the pathogenesis of the lesions cannot be assessed endoscopically.

Both IGH translocations and chromosome 13q deletions are early events in monoclonal gammopathy of undetermined significance and do not evolve during transition to multiple myeloma.

Molecular and genetic events associated with the transition from monoclonal gammopathy of undetermined significance (MGUS) to multiple myeloma (MM) are still poorly characterized. We investigated serial bone marrow specimens from 11 patients with MGUS who eventually progressed to MM (MM post-MGUS) by interphase fluorescence in situ hybridization for immunoglobulin heavy-chain gene (IgH) translocations and chromosome 13q deletions (del(13q)). In nine patients, IgH translocations were present both in MGUS and MM post-MGUS plasma cells, including three t(11;14)(q13;q32) and one t(4;14)(p16;q32), which was observed already 92 months prior to MM. Similarly, all five MM patients with del(13q) had this aberration already at the MGUS stage. Two patients without IgH translocation and del(13q) had chromosomal gains suggesting hyperdiploidy, but IgH translocations and/or del(13q) did not emerge at MM post-MGUS. IgH translocations and del(13q) are early genetic events in monoclonal gammopathies, suggesting that additional events are required for the transition from stable MGUS to progressive MM.

Development of a 13-locus PCR multiplex system for paternity testing.

In this study the development of a 13-locus multiplex-PCR system fitting the updated demands for paternity testing in Germany is described. For this purpose an existing multiplex PCR system that allows the simultaneous amplification of eight different STR loci together with the sex-specific locus amelogenin ( genRESMPX-2, Serac, Germany) was extended. Whereas some of the primers were taken from the underlying multiplex system, suitable primer sequences were chosen for the STR loci D19S433, TPOX, TH01, D16S539, D5S818, D2S1338 and FGA. Primers of loci resulting in potentially overlapping fragment sizes were labelled with the fluorescent dyes 6-FAM, JOE and NED. Reaction conditions, such as annealing temperature, concentrations of primers and polymerase or buffer conditions were optimised to obtain a robust amplification and reproducible genotype analysis for various sample sources. Full DNA profiles from single source samples were reliably typed from template DNA amounts of as low as 120 pg, suggesting a potential use of this system also in forensic casework analysis. With a mean exclusion chance (MEC) of 99.9989% and a power of discrimination (P(D)) of about 1x10(14) (Caucasians), the new multiplex PCR system provides a significant and sensitive system for forensic DNA analysis. On the basis of these studies, a commercial kit system is now provided by Serac (Bad Homburg, Germany, genRESMPX-3).

Sudden death due to a haemoglobin variant.

A previous healthy 35-year-old man was found dead in his truck. Shortly before death he merely complained of influenza-like symptoms. The histological examination revealed evidence of a massive accumulation of sickle cells in smaller blood vessels. After molecular genetic analysis, the preliminary diagnosis of "sickle cell disease" was finally changed to the diagnosis of a sickle cell trait. It is presumed that an epileptic attack which also has to be considered as a concurring cause of death, precipitated sickling of the erythrocytes and led to a fatal sickle cell crisis.

Unusual craniocerebral injury caused by a pneumatic nail gun.

A man was found unconscious near a ladder in a house. After resuscitation he was brought to a hospital and X-rays of the skull showed that two 12-cm long nails had completely penetrated the cranial cavity. The nails were operatively removed and after treatment for 5 weeks, the patient was transferred to a rehabilitation centre with a decreasing hemiparesis on the left side and general deterioration and then, after an attempted suicide to a psychiatric hospital. The perforating cranio-cerebral injury from a pneumatic nail gun known to reach only low muzzle velocities is a very unusual finding.