Magnetic resonance-imaging of the effect of targeted antiangiogenic gene delivery in a melanoma tumour model.

OBJECTIVES: We investigated the effect of targeted gene therapy to melanoma tumours (M21) by MR-imaging. METHODS: M21 and M21-L tumours were grown to a size of 850 mm(3). M21 and M21-L tumours were intravenously treated with an αvß3-integrin-ligand-coupled nanoparticle (RGDNP)/RAF(-) complex five times every 72 hours. MRI was performed at set time intervals 24h and 72h after the i.v. injection of the complex. The MRI protocol was T1-wt-SE±CM, T2-wt-FSE, DCE-MRI, Diffusion-wt-STEAM-sequence, T2-time obtained on a 1.5-T-GE-MRI device. RESULTS: The size of the treated M21 tumours kept nearly constant during the treatment phase (847.8±31.4 mm(3) versus 904.8±44.4 mm(3)). The SNR value (T2-weighted images) of the tumours was 36.7±0.6 and dropped down to 30.6±1.9 (p=0.004). At the beginning the SNR value (T1-weighted images) of the tumours after contrast medium application was 42.3±1.9 and dropped down to 28.5±3.0 (p<0.001). In the treatment group the diffusion coefficient increased significantly under therapy (0.54±0.01x10(-3) mm(2)/s versus 0.67±0.04x10(-3) mm(2)/s). The DCE-MRI showed a reduction of the slope and of the Akep of 67.8±4.3 % respectively 64.8±3.3 % compared to baseline. CONCLUSIONS: Targeted gene delivery therapy induces significant changes in MR-imaging. MRI showed a significant reduction of contrast medium uptake parameters and increase of the diffusion coefficient of the tumours. KEY POINT: • Treatment with targeted gene-delivery therapy can be monitored by MR imaging • DCE and diffusion-weighted imaging are appropriate methods for monitoring this therapy • Functional changes are significant prior to any morphological changes.

Imaging of firefly luciferase activity under antiangiogenic therapy in a heat shock protein 70-transfected melanoma tumor model.

We investigated the effect of targeted gene therapy on heat shock protein 70 (Hsp70) expression in a melanoma tumor model (M21). M21 cells transfected with a plasmid containing the firefly luciferase reporter gene (ffluc), whose expression is driven by the hsp70 (hspa1b) or the cytomegalovirus (CMV) promoter, were grown to a size of 600 mm3. Five animals in each group were intravenously treated with an Arg-Gly-Asp peptide-nanoparticle/Raf-1 kinase inhibitor protein [RGD-NP/RAF(-)] complex. Bioluminescence imaging (BLI) (IVIS, Xenogen, Alameda, CA) was performed at set time intervals. Western blot analysis of the HSP70 protein was simultaneously performed. The size of the treated M21 tumors was nearly constant (637.8 ± 33.4 mm3 vs 674.8 ± 34.4 mm3). BLI showed that if transcription was controlled by the CMV promoter, firefly luciferase activity decreased to 51.1% ± 8.3%. When transcription was controlled by the hsp70 promoter, the highest firefly luciferase activity (4.4 ± 0.3-fold) was observed after 24 hours. In accordance with BLI, Western blot analysis showed an increase in the level of HSP70, with the maximum detection 24 hours after the injection of the RGD-NP/RAF(-) complex. Targeted antiangiogenic therapy can induce luciferase activity where transcription is controlled by an hsp70 promoter and HSP70 protein in melanoma tumors.

Use of in vivo bioluminescence and MRI to determine hyperthermia-induced changes in luciferase activity under the control of an hsp70 promoter.

We investigated the in vivo effect of hyperthermia on the expression of heat shock proteins and MRI changes in three tumor cell lines. Three tumor cell lines (SCCVII, NIH3T3, M21) were transfected with a plasmid containing the heat shock protein 70 gene (hsp70) promoter fragment and the luciferase reporter gene, and injected into mice. Tumors of 1100 mm³ in size were exposed to five different temperatures (38, 40, 42, 44 and 46 °C) in a water bath. Bioluminescence and MRI were performed at set time intervals. The MRI scan protocol was as follows: T1-weighted spin echo ± contrast medium, T2-weighted fast spin echo, dynamic contrast-enhanced MRI, diffusion-weighted stimulated echo acquisition mode sequence, T2 time obtained on a 1.5T General Electric MRI scanner. Immunoblotting was also performed. hsp70 transcription was strongly induced at 42 and 44 °C, reaching values as high as 8531.5 ± 432.1-fold above baseline in NIH3T3 tumors. At these temperatures, significant increases in the uptake of contrast medium, slope of initial enhancement, Ak(ep) values and apparent diffusion coefficient (ADC) were observed in the 8-h scan of the NIH3T3 cell line. In SCCVII tumors, ADC increased by about 23% (p = 0.010) in the scans performed at 8, 24, 48 and 96 h. At 46 °C, luciferase activity was reduced significantly in the three cell lines. In all tumor types, a significant increase in ADC was observed, which was highest in SCCVII tumors (33.8%; p < 0.01). In accordance with the bioluminescence results, significant Hsp70 protein production was shown by immunoblot analysis. The best correlation coefficient between luciferase activity and immunoblotting results was found for M21 tumors (r = 0.93, p < 0.0001). Different tissue types display distinct patterns of hsp70 transcription. MRI can be used, in combination with optical imaging, to provide information on hsp70 transcription and protein production. The major finding of the present study was that heat-related biochemical changes in tumor tissue can be determined by MRI.

Effect of platinum-containing chemotherapy on olfactory, gustatory, and hearing function in ovarian cancer patients.

PURPOSE: Most patients with epithelial ovarian cancer have a poor overall prognosis. Therefore, one of the main therapeutic aims after cytoreductive surgery for these patients is to identify, delay and relieve chemotherapy-induced side effects and optimise the quality of life, especially after first-line therapy. METHODS: Twelve ovarian cancer patients undergoing carboplatinum-containing chemotherapy were assessed using validated tests for olfactory, gustatory, and hearing functions before, during, immediately after, and 3 months after chemotherapy. RESULTS: All chemosensory functions decreased during and after carboplatinum-containing chemotherapy, but recovered 3 months after treatment ended. For olfaction, this decrease was significant, affecting odour identification minimally, and odour threshold the most. For taste, the decrease was not significant, but could be observed in total scores and in each quality (sweet, sour, salty, and bitter). For hearing, the decrease was not significant, but a recovery of the deep and middle frequencies was clearly evident 3 months after chemotherapy. CONCLUSIONS: Patients must be informed about transient declines in chemosensory functions during chemotherapy. Symptomatic relief provided by the use of more spices, a small amount of glutamate, or additional flavouring might help to compensate for decreased functions during chemotherapy and increase patient quality of life.

Evaluation of smell and taste in patients with Wegener's granulomatosis.

Although a reduced olfactory/gustatory function affects patients in all parts of life, this problem has not received much attention in Wegener's granulomatosis (WG). The aim of this study was to assess the smell/taste function of WG patients. Demographic data of 16 WG patients (9 males, 7 females) were obtained. They all subjectively assessed their taste/smell function on visual analogue scale. Olfactory/gustatory functions of the patients were tested with 'Sniffin' Sticks and 'Taste' strips, respectively. The results were then compared with those from sex and age-matched control group (n = 16) and normative data. WG patients subjectively assessed their olfactory (p = 0.03) and gustatory (p = 0.02) function to be lower than control group. All the olfactory scores (odour identification, odour discrimination and threshold) in both genders were significantly below the scores in the control group. WG patients were hyposmic. For taste (total taste score, as well as scores for the qualities sweet, sour, salty and bitter), WG patients did not significantly differ from controls and were normogeusic. However, the gustatory scores showed the tendency of reduction as compared to the control group. In conclusion, WG patients truly suffer from olfactory/taste dysfunction, but this is worse with olfaction. It is, therefore, imperative that physicians should make their patients to be aware of these sensory dysfunctions and educate them on methods to cope with it for better quality of life.

Gene expression analysis of SCC tumor cells in muscle tissue.

The purpose of this study was to evaluate microarray technology of HNSCC cells in muscle tissue. 200 SCCVII tumor cells were injected intramuscularly into the right flank of ten C3H/Km mice each. One week later the animals were killed and the tissue taken out. Histology (H&E staining) and microarray of the tissue were performed. Histology showed a few tumor cells between the muscle fibers. Microarray technology showed different gene expression pattern of the muscle tissue with SCCVII cells in comparison with normal muscle tissue. Only those genes showing a fold change difference of 5 or higher were considered. Gene expression analysis revealed changes in the expression levels of SCCVII cells in muscle tissue in 220 genes. Significant gene expression differences between SCCVII cells in muscle tissue and pure muscle tissue could be seen.

Low flow vascular malformations of the head and neck: a study on brightness mode, color coded duplex and spectral Doppler sonography.

The clinical presentation of low flow vascular malformations of the head and neck (LFVM) can range from a birthmark to severe disfigurement, functional impairment or relevant hemorrhage. The values of Brightness mode (B-mode) ultrasound and Doppler sonography in the investigation, identifying and differentiating of these lesions has been sparingly documented in the literature. This study evaluates the sonografic features of different morphological subtypes of LFVM. This is a 2-year retrospective study of 51 patients who presented with LFVM based on routine ultrasound exam in the context of their clinical consultation. Diagnosis was based on the clinical and histological findings. B-mode, color coded duplex and spectral Doppler measurements were performed for venous, lymphatic, capillary, and mixed venous-lymphatic lesions of the head and neck. The echogenicity of the majority of venous malformations was heterogenic, of most lymphatic malformations hypoechoic, and of all capillary malformations isoechoic. Blood flow was detected in only 11 cases (36.7%) of venous malformations with a monophasic pattern. There was a statistical significant difference in the mean minimum and maximum Doppler shifts between venous and lymphatic malformation for cases when the blood flow was evident. No statistical significant difference in Doppler parameters existed between capillary and lymphatic, neither between venous and capillary nor mixed malformations. Phleboliths were present in eight cases (26.7%) of venous malformations and were not detectable in any other subtype of LFVM. The detection of flow in ultrasound was only possible in a small portion of LFVM. When considering differentiating among LFVM, features such as the echogenecity, spectral Doppler wave forms, and the evidence of phleboliths contribute to establish the correct diagnosis.

Gustatory and olfactory function in breast cancer patients.

PURPOSE: Loss of appetite is a common complaint in cancer patients. There is still no overall conclusion whether this symptom might be caused by distorted taste/smell function or by tumor byproducts. This knowledge would be important for adequate patient counseling as well as symptom relief. Several studies investigated taste function, but to our knowledge, only one studied olfactory function in cancer patients. MATERIALS AND METHODS: Sixty-nine breast cancer patients were investigated by a validated taste (taste strips) and smell test (Sniffin' Sticks) prior to chemotherapy. RESULTS: Compared to normative data, breast cancer patients showed no significant difference in odor threshold, but better scores for odor identification and odor discrimination. For taste, breast cancer patients showed a significantly lower value for the quality sour compared to healthy controls only on left side of the tongue; there was no difference in the qualities sweet, salty, and bitter. An increase in tumor size was associated with a significant decrease in olfactory function, but not in gustatory function. Different histology or graduation of breast cancer, resection status, or metastasis to the lung and liver had no influence on taste and smell. There was no correlation between taste and smell to estrogen or progesterone receptor status. There was no correlation between smell and human epidermal growth factor receptor 2 (Her2 status), but there was a significant correlation between bitter taste and Her2 status. CONCLUSION: Taste/smell did not seem substantially altered in breast cancer patients compared to normative data. Nevertheless, increasing tumor burden resulted in decreased olfactory function, but not in taste changes.

Olfactory and gustatory function in irritable bowel syndrome.

Irritable bowel syndrome (IBS) is the most common, functional disorder diagnosed by gastroenterologists. It is still unclear whether IBS has a central etiology, e.g., hyperreactivity of the brain, or a peripheral etiology, e.g., stimulation of olfactory/gustatory receptors on enterochromaffin cells, followed by serotonin release and changed gut motility. Testing the odor identification (ID), odor discrimination (DIS) and odor threshold (THR) as well as the total taste and the taste qualities "sweet", "sour", "salty" and "bitter" should be of help for determining the etiology. To our knowledge, this is the first study investigating the olfactory/gustatory function in IBS patients. The olfactory/gustatory function of 43 patients (32 women, 11 men) suffering from IBS as defined by the ROME III criteria was investigated by means of validated tests (Sniffin' Sticks and taste strips). Compared to normative data, scores of THR were decreased and scores of ID and DIS were increased in IBS patients. Additionally, when compared to normative data, there was no difference in the taste function of IBS patients. Assuming that THR reflects more the peripheral olfactory function, whereas ID and DIS are influenced by central activity, and that taste did not differ in IBS patients compared to normative data, this supports the idea of a central etiology of IBS.

Effect of continuous high intensity focused ultrasound in a squamous cell carcinoma tumor model compared to muscle tissue evaluated by MRI, histology, and gene expression.

The purpose of this study was to investigate the effect of the continuous mode of high intensity focused ultrasound (HIFU) in a mouse head and neck cancer model (SCCVII) compared to muscle tissue. HIFU was applied to SCCVII tumors and to muscle tissue in C3H/Km mice using a dual ultrasound system (imaging 6 MHz/therapeutic 1 MHz). A continuous HIFU mode (total time 20 sec, intensity 6730.6 W/cm(2)) was applied. Three hours after HIFU treatment pre- and post-contrast T1-wt, T2-wt images, and a diffusion-wt STEAM sequence were obtained. After MR imaging, the animals were euthenized and the treated tumor and muscle tissue was taken out for histology and functional genomic analysis. T2 images showed increased signal intensity, post-contrast T1 showed a decreased contrast uptake in the central parts in the tumor tissue as well as in the muscle tissue. In addition a significant higher diffusion coefficient was found in both tissue types. Histological evaluation (H&E, Immunohistochemistry) of the tumors and the muscle tissue revealed areas of significant necrosis. In the tumor tissue 23 genes were up-regulated (> 2 fold change) and 4 genes were down-regulated (< -2 fold change). In the muscle tissue 29 genes were up-regulated and 17 genes down-regulated. Thirteen genes were up-regulated in both tissue types, 8 genes only in the SCCVII tissue, and 11 genes only in the muscle tissue. The use of HIFU treatment on tumor and muscle tissue results in dramatic changes in gene expression. The expression of some genes are tissue specific, the expression of other genes are independent of the tissue type.

Gene expression profiles, histologic analysis, and imaging of squamous cell carcinoma model treated with focused ultrasound beams.

OBJECTIVE: The purpose of our study was to evaluate the effect of short-pulse high-intensity focused ultrasound (HIFU) on inducing cell death in a head and neck cancer model (SCCVII [squamous cell carcinoma]) compared with continuous HIFU to get a better understanding of the biologic changes caused by HIFU therapy. MATERIALS AND METHODS: HIFU was applied to 12 SCCVII tumors in C3H/Km mice using a dual sonography system (imaging, 6 MHz; therapeutic, 1 MHz). A continuous HIFU mode (total time, 20 seconds; intensity, 6,730.6 W/cm2) and a short-pulse HIFU mode (frequency, 0.5 Hz; pulse duration, 50 milliseconds; total time, 16.5 minutes; intensity, 134.4 W/cm2) was applied. Three hours later, MR images were obtained on a 1.5-T scanner. After imaging, the treated and untreated control tumor tissue samples were taken out for histology and oligonucleotide microarray analysis. RESULTS: Prominent changes were observed in the MR images in the continuous HIFU mode, whereas the short-pulse HIFU mode showed no discernible changes. Histology (H and E, TUNEL [terminal deoxynucleotidyl transferase-mediated dUTP {deoxyuridine triphosphate} nick-end labeling], and immunohistochemistry) of the tumors treated with the continuous HIFU mode revealed areas of significant necrosis. In the short-pulse HIFU mode, the H and E staining showed multifocal areas of coagulation necrosis. TUNEL staining showed a high apoptotic index in both modes. Gene expression analysis revealed profound differences. In the continuous HIFU mode, 23 genes were up-regulated (> twofold change) and five genes were down-regulated (< twofold change), and in the short-pulse HIFU mode, 32 different genes were up-regulated and 16 genes were down-regulated. CONCLUSION: Genomic analysis might be included when investigating tissue changes after interventional therapy because it offers the potential to find molecular targets for imaging and therapeutic applications.

In vitro effect of focused ultrasound or thermal stress on HSP70 expression and cell viability in three tumor cell lines.

RATIONALE AND OBJECTIVES: In this study, we compared the effect of focused ultrasound with the effect of thermal stress on the induction of a heat inducible promoter in an in vitro model using three tumor cell lines (M21, SCCVII, and NIH3T3). MATERIALS AND METHODS: We used a reporter construct that was generated using the stress-inducible promoter from the gene encoding a murine 70-kilodalton heat shock protein (Hsp70A.1) and a luciferase (luc) reporter plasmid. High-intensity focused ultrasound (HIFU) was applied in two different modes. In the first mode, an increasing voltage at constant pulse duration and in the second mode a constant voltage at increasing pulse duration was applied. HIFU or thermal stress was delivered over a range of temperatures (36-52 degrees C) for 5 minutes, and resulting luciferase activity was measured in live cells using a cooled charge-coupled device camera as a measure of reporter gene transcription. Luciferase activity was measured at set time intervals for a total of 108 hours post-stress. RESULTS: Both methods induced the hsp70 promoter; however, the luciferase activity under the influence of HIFU, independent of the applied mode, and thermal stress differs despite the fact that the temperature was the same. In the M21 tumor cell line, the maximum luciferase activity after focused ultrasound application was 4818 +/- 1521% at a temperature of 48 degrees C and after thermal stress 4468.2 +/- 1890.2% at a temperature of 52 degrees C with a viability of 72.3 +/- 5.2% and 85 +/- 3.4%, respectively. In the SCC tumor cell line, the maximum luciferase activity after focused ultrasound application was 6743.0 +/- 3281.4% and after only thermal stress exposure was 3910.6 +/- 2189.0% at a temperature of 44 degrees C and 50 degrees C, respectively. At the highest luciferase activity, the portion of vital cells was 72.5 +/- 8.4% and 72.5 +/- 5.9% respectively. In the NIH3T3 tumor cell line the highest luciferase activity of 428510.6 +/- 26526.8% was seen at a temperature of 42 degrees C applying focused ultrasound. Under thermal stress it was 29221.3 +/- 7205.0% at a temperature of 50 degrees C. At the highest luciferase activity, the viability analysis showed 75.3 +/- 9.2% and 72.3 +/- 7.9% viable cells, respectively. CONCLUSIONS: Focused ultrasound induces hsp70 expression like thermal stress alone; however, HIFU is capable of inducing expression at lower temperatures than heat stress alone, indicating that nonthermal effects also play a role on the induction of hsp70.

Initial Experience with the Transapical Access for TEVAR.

Background The endovascular approach has become a mainstay in the treatment of aortic aneurysms. While the transfemoral approach is most commonly used, it is often inaccessible due to a vascular pathology, such as occlusion, tortuosity or heavy calcifications. The transapical access provides an alternative approach. The goal of this study is to report the feasibility of the transapical approach for total endovascular repair of thoracic aortic aneurysms (TEVAR). Methods Three patients with thoracic aortic aneurysms with inaccessible femoral arteries underwent TEVAR via the transapical approach. For access, the apex of the left ventricle was exposed by a mini-thoracotomy. After left ventricular puncture, a stiff guidewire was placed through the aortic valve into the descending thoracic aorta. All stent grafts were delivered under fluoroscopic guidance. Results All three procedures were technically successful with complete exclusion of the aneurysm without endoleak. One patient suffered spinal ischemia with subsequent paraplegia on day 4 after the procedure. The same patient died on day 43 after the procedure due to esophageal rupture. The two remaining patients did not present procedure-related problems. Conclusion The transapical access is a feasible and safe alternative to the transfemoral route in selected cases scheduled for TEVAR. Key Points · Transapical TEVAR is a feasible option if the transfemoral route is not accessible.. · Reversed orientation of the stent graft has to be taken into account for procedure planning and graft deployment.. · Care has to be taken in the case of aortic valve vegetation in order to avoid stroke due to downstream embolism.. Citation Format · Mahnken AH, Irqsusi M, Hundt W et al. Initial Experience with the Transapical Access for TEVAR. Fortschr Röntgenstr 2017; 189: 760 - 764.

Value of PET/CT versus PET and CT performed as separate investigations in patients with Hodgkin's disease and non-Hodgkin's lymphoma.

PURPOSE: The aim of this study was to assess the clinical benefit of combined [(18)F]FDG PET/CT in patients with malignant lymphoma as compared to separately performed PET and CT. METHODS: Overall, 100 patients with Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) were included in this study. Co-registered PET/CT with [(18)F]FDG and contrast medium was performed in 50 consecutive patients with NHL (n=38) or HD (n=12) for initial staging (IS) (n=12) or re-treatment staging (RS) (n=38). Another 50 patients with NHL (n=32) or HD (n=18) underwent separate PET and CT investigations within a time frame of 10 days for IS (n=22) or RS (n=28). Lymphoma involvement was separately evaluated for seven different regions in each patient. Each patient had clinical follow-up evaluation for >6 months. PET and CT data were analysed separately as well as side-by-side or in fused mode. RESULTS: In the PET/CT group, region-based evaluation for lymphoma involvement suggested a sensitivity/specificity of 85%/91% for CT, 98%/99% for PET and 98%/99% for PET/CT. In the PET and CT group, region-based evaluation showed a sensitivity/specificity of 87%/80% for CT, 98%/99% for PET and 98%/100% for PET and CT read side by side. CONCLUSION: PET was superior to CT alone and was improved further by side-by-side reading of both examinations. However, no significant difference was observed between PET/CT and separate PET and CT imaging in patients with lymphoma.

Smell and taste in patients with vascular malformation of the extracranial head and neck region.

BACKGROUND: Olfactory and gustatory functions have not been investigated in patients with vascular malformation of the extracranial head and neck region with validated smell and taste tests. Although olfactory and gustatory deficiencies are often not outwardly apparent, they substantially affect daily life. METHODS: Smell and taste tests using sniffin sticks and taste strips were administered in 40 patients. RESULTS: For all age groups and both sexes, odor threshold (THR) values were, on average, lower in patients than in healthy individuals; whereas, values of odor identification and discrimination were not significantly lower. Regarding odor THR, 33 (82.5%) patients were hyposmic. Taste values (sweet, sour, salty, bitter, and total taste) were, on average, lower in patients than in healthy individuals; 21 (52.5%) patients were hypogeusic. Disease duration did not correlate with smell and taste test values. Patients with and without tongue involvement had decreased odor threshold and taste values. No significant differences were identified when taste values on the left and right sides of the tongue were compared in patients without tongue involvement and with unilateral and bilateral tongue involvement. Patients with venous malformations had lower smell test values, and patients with lymphatic malformations had lower taste test values than patients with other malformations. CONCLUSION: Patients exhibit significantly reduced olfactory and gustatory function even when the nose and/or tongue are not malformed. Patients should be tested with validated smell and taste tests to adequately inform and advise them about overcoming smell and taste deficits.

Smell and taste in patients with vascular malformation of the extracranial head and neck region.

BACKGROUND: Olfactory and gustatory functions have not been investigated in patients with vascular malformation of the extracranial head and neck region with validated smell and taste tests. Although olfactory and gustatory deficiencies are often not outwardly apparent, they substantially affect daily life. METHODS: Smell and taste tests using sniffin sticks and taste strips were administered in 40 patients. RESULTS: For all age groups and both sexes, odor threshold (THR) values were, on average, lower in patients than in healthy individuals; whereas, values of odor identification and discrimination were not significantly lower. Regarding odor THR, 33 (82.5%) patients were hyposmic. Taste values (sweet, sour, salty, bitter, and total taste) were, on average, lower in patients than in healthy individuals; 21 (52.5%) patients were hypogeusic. Disease duration did not correlate with smell and taste test values. Patients with and without tongue involvement had decreased odor threshold and taste values. No significant differences were identified when taste values on the left and right sides of the tongue were compared in patients without tongue involvement and with unilateral and bilateral tongue involvement. Patients with venous malformations had lower smell test values, and patients with lymphatic malformations had lower taste test values than patients with other malformations. CONCLUSION: Patients exhibit significantly reduced olfactory and gustatory function even when the nose and/or tongue are not malformed. Patients should be tested with validated smell and taste tests to adequately inform and advise them about overcoming smell and taste deficits.

Detection of micrometastases of squamous cell carcinoma tumor cells in muscle tissue.

The aim of this study was to evaluate microarray technology in the detection of micrometastases of head and neck squamous cell carcinoma (HNSCC) in muscle tissue. Three hundred SCCVII tumor cells were injected intramuscularly into the right flank of ten C3H/Km mice. One week later, the animals were euthanized and the muscle tissue was taken out. Histology (H&E staining), microarray and reverse transcriptase polymerase chain reaction analysis (RT-PCR) of the tissue was performed. Histology showed a few tumor cells between the muscle fibers. Microarray technology showed the different gene expression pattern of the muscle tissue with micrometastases in comparison to normal muscle tissue. Only genes with a fold change difference of 10 or greater were considered. Gene expression analysis revealed changes in the expression levels of 91 genes of micrometastases in muscle tissue. RT-PCR confirmed gene up-regulation. Significant differences in gene expression between micrometastases in muscle tissue and pure muscle tissue were found. The genes found to be up-regulated could be used to detect micrometastases in muscle tissue.

Smell and taste in inflammatory bowel disease.

OBJECTIVE: To investigate the olfactory/gustatory functions of patients with inflammatory bowel disease (IBD) by smell/taste tests, and to determine if disease activity or medication might influence the olfactory/gustatory functions of patients. PATIENTS AND METHODS: In total, 59 IBD patients (37 Crohn's disease (CD) and 22 ulcerative colitis (UC) patients) were studied using "Sniffin' sticks" and "taste strips" for olfactory and gustatory tests, respectively, and compared to healthy controls and published normative data. RESULTS: Among IBD (CD and UC) patients, the values for odor threshold, but not for odor identification or discrimination, were significantly lower than that of the normative data. Further, these patients showed lower values than the normative taste values and the control group for all tastes, except sour; 57.6% of the IBD patients were hyposmic, while 30.5% were hypogeusic. Subjective self-assessments showed that the patients were not aware of their reduced olfactory/gustatory functions. There were no relevant differences in taste and smell abilities between the CD and UC patients. Disease activity and treatment did not influence the olfactory/gustatory functions. CONCLUSION: IBD (CD and UC) patients exhibited significant reductions in the olfactory and gustatory functions. Therefore, patients should be tested by smell/taste tests, in order to be adequately informed of their olfactory/gustatory functions and provided an understanding of how to overcome their limitations, and thus improve their quality of life.

Combined treatment of subacute and acute synthetic and venous bypass-graft occlusions with percutaneous mechanical thrombectomy and thrombolysis.

INTRODUCTION: Percutaneous mechanical thrombectomy (PMT) is a third choice of treatment for acute arterial occlusions, in addition to thrombolysis and surgical thrombectomy. The aim of this retrospective study was to compare the combined treatment of PMT and local thrombolysis with thrombolysis therapy alone. MATERIALS AND METHODS: Sixty-nine patients with acute (<14 days [n = 35]) or subacute (14-42 days [n = 34]) femoropopliteal bypass occlusions were treated with PMT combined with thrombolysis. Seventy-two patients with acute [n=40] or subacute [n = 32] femoropopliteal bypass occlusions were treated with thrombolysis alone. The thrombolysis in myocardial infarction (TIMI) classification was used to assess the bypass occlusion. Local thrombolysis time and dosage, reopening time, time in the intensive care unit, necessary surgical re-interventions, and clinical outcome were compared between the 2 groups. RESULTS: The TIMI scores were significantly higher in the PMT plus thrombolysis group than in the thrombolysis group (acute occlusions 1188 versus 935, p<0.001; subacute occlusions 935 versus 605, p<0.001). The total urokinase dosage, the total hours of thrombolysis, time in the intensive care unit, and total hospital stay in the acute PMT plus thrombolysis group were significantly lesser than those in the thrombolysis group. After 24h of treatment, the ankle-brachial index improved in all groups (p<0.001): in the acute and subacute PMT plus thrombolysis group to 0.63 ± 0.14 and 0.43 ± 0.08, respectively; and in the acute and subacute thrombolysis group to 0.51 ± 0.11 and 0.41 ± 0.04, respectively. CONCLUSIONS: PMT combined with thrombolysis is a safe and very effective therapy for acute and subacute femoropopliteal bypass occlusions compared to treatment with thrombolysis alone.

Is routine audiometric evaluation necessary in gynaecologic tumour patients undergoing chemotherapy?

BACKGROUND: Our objective was to assess the auditory function of gynaecological tumour patients who had received cytotoxic agents and to determine their associated risk of ototoxicity. PATIENTS AND METHODS: 87 patients who had undergone chemotherapy for gynaecological malignancies were investigated. Of these patients, 79% had breast cancer, and 14% ovarian cancer. All of the patients had a subjective assessment of their hearing function on a visual analogue scale. Audiometric tests were performed before and at 9 weeks, 18 weeks and 3 months after completion of chemotherapy. RESULTS: The age of the patients ranged from 32 to 71 years (mean age of 53.5 ± 10.5 years). The average subjective rating of the patients' hearing function was 83.0 ± 17.2 before and 84.8 ± 16.9 3 months after completion of chemotherapy. No significant audiometric change at either the speech hearing frequency range (0.5-2 KHz) or high frequencies was observed in the patients after chemotherapy. There was also no significant difference in the hearing threshold of the patients who had received platinum analogue-based chemotherapy compared to non-platinum analogue-based chemotherapy. CONCLUSION: Hearing loss is uncommon in patients treated with the typical gynaecological chemotherapy protocols. Hence, routine audiometric testing in these patients is not necessary.