[A systematic review and current recommendation for treatment of rhinophyma]. / Übersicht und aktuelle Empfehlung zur Therapie des Rhinophyms.

Rhinophyma is a benign disease involving the skin of the nose, which is characterized by skin thickening and hypertrophy of the sebaceous glands and connective tissue. It occurs predominantly by Caucasians in their fifth to seventh life decades and is 12-30 times more likely to affect males. The etiology and pathogenesis of Rhinophyma remains unclear, however it is generally stated to be the final stage of rosacea. A causative relationship between rhinophyma and alcohol consumption has not yet been proven. This review highlights the treatment options of rhinophyma. Surgery is gold standard for management of advanced rhinophyma. Each technique has pros and cons, but using surgical instruments with monopolar energy as loop and ball electrode seem to combine several goals best - first of all simultaneous removal and hemostasis followed by nasal contour refinement. Due to possible coincidence of skin cancer such as a basal- or squamous cell carcinoma, histopathological examination of all removed tissue is recommended.

Modified Nucleosides - Molecular Markers Suitable for Small-volume Cancer?

BACKGROUND: Modified nucleosides (mNS) in urine are shown to be encouraging markers in cancer, mostly in patients presenting with high tumor mass such is breast and lung cancer. To our knowledge, mNS have not been investigated in head and neck squamous cell carcinoma (HNSCC). HNSCC is characterized by early metastasis into locoregional lymph nodes and slow infiltrating growth, but even in the advanced stage exhibits only a relatively low cancer volume. Therefore, reliable distinction between HNSCC and healthy controls by urinary mNS might pose substantial analytical problems and even more as patients with HNSCC mostly have an increased exposure to tobacco smoke and excessive alcohol consumption which affect the renal mNS pattern. MATERIALS AND METHODS: Urinary mNS in samples of 93 therapy-naive patients with HNSCC and 242 healthy controls were quantified by reversed-phase high-performance liquid chromatography. Considering that the circadian rhythm causes diuresis-induced variations in concentration, the mNS-to-creatinine ratio was chosen to compare patients and controls. For sensitivity and specificity in discriminating between patients and controls, the corresponding curve was plotted. Additionally, logistic regression was carried out and a multilayer perceptron neuronal network (NN) was created. RESULTS: Fifteen mNS were detectable in cases and controls; concentrations of 11 were found to be significantly different. The sensitivity and specificity depend on the total volume of the lesion; HNSCC with volume <20 ml was reliably detected, but those with a volume of 20 ml or greater produced amounts of mNS which led to the most accurate detection of HNSCC based on HNSCC-specific mNS patterns. CONCLUSION: Analysis of urinary mNS allows for detection of small-volume HNSCC, with acceptable specificity and sensitivity if the tumor volume exceeds 20 ml.

Functional and cosmetic donor site morbidity of the radial forearm-free flap: comparison of two different coverage techniques.

BACKGROUND: The use of the radial forearm-free flap is a well-established and reliable reconstruction method in head and neck surgery. Usually, the defect of the donor site is covered with full or split-thickness skin grafts. Since 09/2013, a direct closure of the radial forearm donor site has been performed at the ENT University Hospital Leipzig to avoid secondary donor site morbidity. However, few data are available in the literature on long-term cosmetic and functional results compared to the established indirect donor site defect coverage. METHODS: This study investigated patients with radial forearm-free flap harvest from 01/2012 until 03/2015. A total of n = 39 patients were included, with n = 18 being operated by indirect (group 1) and n = 21 by direct closure technique (group 2). For the validation of surgical revisions and wound healing disorders, we carried out clinical investigations as well as interviews. The "POSAS Observer and Patient Scale" was used for assessing the cosmetic outcome and the "Michigan Hand Outcome Questionnaire (MHQ)" for functional criteria. RESULTS: Group 2 showed an increased rate of wound healing problems, however it was not statistically different compared to group 1. Revision surgery was necessary in both groups only each in one case. Using the POSAS, there were no significant differences between both groups in the observer scale for the items vascularity, pigmentation, thickness, relief, pliability, surface area and even for pain, scar itching, color, stiffness, thickness and relief in the patient scale. The functional results (MHOQ) also showed no significantly inferior results for group 2. CONCLUSIONS: The direct closure procedure is quick, simple and can be performed without secondary donor site morbidity. For wound healing, cosmetic and function of the forearm and hand, no inferior results can be measured for the direct procedure compared to the indirect coverage technique.

HLA traits linked to development of head and neck squamous cell carcinoma affect the progression-free survival of patients.

BACKGROUND: Personalized medicine and treatment stratification of patients with head and neck squamous cell carcinoma (HNSCC) today mostly ignore genetic heterogeneity in HNSCC but especially the patient's genetic background. We hypothesized that particular human leukocyte antigen (HLA) class I (HLA-A, B, Cw) and II proteins (DR, DQ) confer susceptibility for and influence development of HNSCC and may be prognostic factors for progression-free survival (PFS). METHODS: 90 consecutive HNSCC patients of the prospective observational cohort study LIFE treated between 08/2010 and 05/2011 at the University Leipzig underwent low resolution typing of HLA-A, B, Cw, DR, and DQ. Antigen and haplotype frequencies were compared to those in German blood donors. Effects on PFS were analyzed using Kaplan-Meier curves and Cox models. RESULTS: HNSCC patients had overall altered HLA-B frequencies (P<0.05); frequencies of B∗44 were lower, those of B∗13, B∗52, and B∗57 increased (P<0.05). Almost all other antigen frequencies showed no deviation. Homozygous HLA-Cw and DRB4 were frequent and associated with reduced PFS (P<0.05). Altered haplotype frequencies were common and particular haplotypes accompanied by differing PFS. B∗13/Cw∗06 carriers had poorest outcome (P=0.011). However, multivariate Cox proportional hazard models revealed 3 clinical covariates (localization oropharynx, loco-regional metastasis, and T4 category), HPV16-DNA positivity, and 10 HLA traits as independent predictors for PFS. CONCLUSIONS: The relevance of the genetic background of HNSCC patients calls for future research to clarify the role of HLA traits in HNSCC and if PFS depends on HLA.

Pediatric nasolacrimal duct obstruction-benefit of a combined therapeutic approach.

BACKGROUND: Pediatric nasolacrimal duct obstruction (PNDO) requires therapeutic intervention after conservative procedures failed. As resilient treatment guidelines for the treatment are missing, the aim of this study was to evaluate the advantages of two different intervention techniques in children with PNDO. METHODS: Between January, 2006 and June, 2014, 233 children (0-208 months) were treated either with conventional probing by ophthalmologists only (Group I) or with endonasal endoscopic interdisciplinary approach (Group II). The clinical outcome was analyzed. RESULTS: The overall success rate of Group I was 93.4% compared to 98.4% of Group II (P<0.05). 50% of all interventions (n=62) of Group II required further surgical procedures in addition to probing/irrigation, particularly with regard to children <6 and >24 months. CONCLUSIONS: Endoscopic control in treatment of PNDO allows exact identification of the stenosis and appropriate surgical intervention with an improved clinical outcome. Endonasal endoscopic surgical techniques should be the standard PNDO treatment.

Reduced proliferation and colony formation of head and neck squamous cell carcinoma (HNSCC) after dual targeting of EGFR and hedgehog pathways.

PURPOSE: The hedgehog signalling pathway (Hh) is frequently active in head and neck squamous cell carcinoma (HNSCC). Overexpressed Hh associates with poor prognosis. The Hh inhibitor vismodegib targets smoothened, and based on molecular data, may prevent resistance to EGFR targeting. METHODS: To elucidate potential roles of vismodegib in HNSCC therapy, its sole effects and those combined with cisplatin, docetaxel, and cetuximab on HNSCC cell lines were assessed by MTT metabolisation and BrdU incorporation. Colony formation (CF) of primary HNSCC cells was studied utilizing the FLAVINO-protocol. Combinatory effects were analysed regarding antagonism, additivity or synergism. Associations between the ex vivo detected mode of action of vismodegib with other treatments related to patient characteristics were assessed and progression-free survival (PFS) in patient groups compared using Kaplan-Meier curves. RESULTS: Vismodegib suppressed BrdU incorporation significantly stronger than MTT turnover; CF was significantly inhibited at ≥20 µM vismodegib while concentrations <20 µM acted hormetic. Combining 20 µM vismodegib plus docetaxel (T), cisplatin (P), and cetuximab (E), additively enhanced anti-tumoral activity in HNSCC samples from patients with superior PFS highlighting a potential role for ex-vivo testing of this combination for use as a prognostic classifier. CONCLUSION: We provide ex-vivo evidence for vismodegib's potential in HNSCC therapies, especially if combined with cetuximab, cisplatin and docetaxel.

Stimulation by Monocyte Chemoattractant Protein-1 Modulates the Ex-vivo Colony Formation by Head and Neck Squamous Cell Carcinoma Cells.

BACKGROUND: The outcome of patients with head and neck squamous cell carcinoma (HNSCC) is still poor. To improve therapy of HNSCC, biomarkers indicating progression of the disease or modifiers with potential as therapeutic targets and therapy need to be investigated. Since monocyte chemoattractant protein (MCP1) is potentially involved in tumorigenesis of HNSCC, we aimed to clarify its role in HNSCC and investigated the influence of stimulation by MCP1 and its depletion using antibodies against MCP1 (anti-MCP1) on colony formation by HNSCC cells. MATERIALS AND METHODS: Biopsies of HNSCC were treated according to the protocol of the FLAVINO assay with cisplatin, docetaxel, temsirolimus or cilengitide alone, or combined with MCP1 or anti-MCP1. After a 72-h incubation, ethanol-fixed and fluoresceine-isothiocyanate (FITC)-labeled epithelial colonies were counted. RESULTS: Colony formation was significantly suppressed by MCP1 and 3.3 µM cisplatin, while docetaxel, cilengitide and temsirolimus at concentrations of 0.275, 10 and 0.50 µM caused insignificant effects. Addition of MCP1 to cisplatin, docetaxel and cilengitide increased efficacy of cytostatics in inhibition of colony formation, whereas those with temsirolimus were increased by anti-MCP1 that when applied alone failed to modulate colony formation. Overall regarding facilitated chemosensitivity, there was a statistical trend in favor of MCP1 stimulation over depletion. CONCLUSION: Our ex vivo results show context-dependent effects of MCP1 in HNSCC cells. An increase of MCP1 level or its addition to cisplatin, docetaxel and cilengitide reduce colony formation but the efficacy of temsirolimus is augmented by MCP1 depletion. These context-dependently opposite outcomes call for further translational investigations in HNSCC.

Simvastatin suppresses head and neck squamous cell carcinoma ex vivo and enhances the cytostatic effects of chemotherapeutics.

PURPOSE: Simvastatin (Sim) is approved as lipid-controlling drug in patients with cardiovascular risk to reduce hypercholesterolemia. Recent publications indicate possible inhibitory effects of Sim on tumor cell lines, and epidemiological data suggest activity in cancer patients. Still, its therapeutic efficacy, particularly in head and neck squamous cell carcinoma (HNSCC), remains to be elucidated. This study analyzes the effects of Sim on HNSCC cell lines (KB, HN5, FaDu) and on a larger set of primary HNSCC cells by employing a short-time ex vivo colony formation test (FLAVINO assay). Possible additive or synergistic effects of Sim combinations with established chemotherapeutics are determined as well. METHODS: Biopsies of 49 HNSCC were tested in the FLAVINO assay with Sim alone or in combination with cisplatin (Cis) or docetaxel (DTX). Cell lines were studied for reference. Epithelial HNSCC cells were stained by Cy2-labeled anti-cytokeratin antibodies facilitating the detection of colony formation (CF) by immunofluorescence. Drug combinations were analyzed regarding their interaction. RESULTS: Sim alone acted suppressive on tested cell lines and increased the cytostatic efficacy of Cis and DTX. 18/49 HNSCC qualified for FLAVINO-based dose-response analyses, and Sim significantly suppressed CF in 18/18 primary HNSCC. Moreover, Sim increased cytotoxic effects of Cis and DTX, primarily in an additive mode of action. CONCLUSIONS: The ex vivo tumor cell inhibition of Sim and its additive effects upon combination with established cytostatics provide the basis for epidemiological and clinical studies on statins, potentially directed toward co-medication in future treatment regimens.

Endonasal nasolacrimal duct surgery: a comparative study of two techniques.

Dacryocystorhinostomy (DCR) is performed in patients with saccal or postsaccal lacrimal duct obstruction. Focusing on the endonasal approach, we compared success rates, clinical outcome, complications and patient satisfaction of endoscopic vs. non-endoscopic techniques in endonasal DCR. We analyzed the results of 173 patients who underwent an endonasal DCR either utilizing a non-endoscopic (Group I) or an endoscopic technique (Group II) between 2006 and 2011. Irrigation tests followed the first day and at least 3 months after surgery. Postoperative patients' satisfaction and the occurrence of symptoms were documented and evaluated in a follow-up questionnaire. The minor complication rates of both endonasal DCR techniques were similarly low (10%) without severe adverse events. The use of the endoscope prolonged the operating time significantly (28 ± 9 min Group I vs. 34 ± 14 min Group II, p = 0.003). The success rate was 90.2% in Group II compared to only 57.9% in Group I (p < 0.000). Further, we determined the absence of reflux during the irrigation test 1 day after surgery as a significant predictor for the later outcome (R = 1.598, p = 0.005). The follow-up questionnaire revealed a significant improvement in subjectively perceived symptoms by the surgical intervention for both endonasal techniques (p < 0.000). The endoscopically assisted DCR is a safe and successful endonasal technique for patients with saccal or postsaccal lacrimal duct obstructions. The use of the endoscope led to significant higher success rates compared to non-endoscopic techniques in our collective.

Hedgehog targeting by cyclopamine suppresses head and neck squamous cell carcinoma and enhances chemotherapeutic effects.

BACKGROUND: The hedgehog signaling pathway (HH) is involved in tumorigenesis in a variety of human malignancies. In head and neck squamous cell carcinomas (HNSCC), Hh overexpression was associated with poor prognosis. Therefore, we analyzed the effect of Hh signaling blockade with cyclopamine on colony formation of cells from HNSCC samples. PATIENTS AND METHODS: HNSCC biopsies were cultured alone for reference or with serial dilutions of cyclopamine (5-5,000 nM), docetaxel (137.5-550 nM), or cisplatin (1,667-6,667 nM) and their binary combinations. Cytokeratin-positive colonies were counted after fluorescent staining. RESULTS: Cyclopamine concentration-dependently inhibited HNSCC ex vivo [(IC50) at about 500 nM]. In binary combinations, cyclopamine additively enhanced the suppressive effects of cisplatin and docetaxel on HNSCC colony formation. CONCLUSION: Our findings define SMO--a Hh component- as a potential target in HNSCC and suggest the utility of Hh targeting in future multimodal treatment regimens for HNSCC.

Overexpression of the Hedgehog signalling pathway in head and neck squamous cell carcinoma.

BACKGROUND: Overexpression of the Hedgehog (HH) signalling pathway has been described in several malignancies and is associated with a poor prognosis. HH signalling blockade reduces tumour growth in vitro and in vivo. We aimed to determine whether head and neck squamous cell carcinomas (HNSCCs) express HH proteins in comparison to healthy mucosa. PATIENTS AND METHODS: Formalin-fixed and paraffin-embedded tissue sections of 10 patients with HNSCC were stained with fluorescence-labelled antibodies for cytokeratin and HH proteins (SHH, PTCH1/2, SMO, Gli1-3) and photographs were taken with a laser scanning microscope. The pixel count and colour intensity were analysed in RGB (red/green/blue) colour mode, and expression levels were compared to healthy mucosa. RESULTS: Image analysis in RGB mode provided objective evidence for the over-expression of HH signalling components in HNSCC, particularly with regard to the transcription factors Gli1 (10-fold) and SHH (5-fold) in comparison with healthy mucosa. The lowest levels were found for Gli3 in HNSCC. CONCLUSIONS: We postulate pivotal roles of Gli1 and SHH expression in the carcinogenesis of HNSCC. HH pathway overexpression appears to be involved in the initiation of tumour growth and spread due to its stem cell-modulating properties. Detection of HH pathway components, and especially Gli1 and SHH, in HNSCC might offer a promising target for further anticancer research in HNSCC.

Combined effects of lapatinib and cisplatin on colony formation of head and neck squamous cell carcinoma.

BACKGROUND: Lapatinib targets human epidermal growth factor-receptor (EGFR) and Her2/neu receptor tyrosine-kinases and hence is under investigation in multimodal therapy concepts of advanced head and neck squamous cell carcinoma (HNSCC). We studied combined effects of lapatinib and cisplatin on colony formation (CF) of epithelial cells of individual HNSCC in short term ex vivo assays. MATERIALS AND METHODS: Biopsies of HNSCC were minced, collagenase-digested and exposed to serial lapatinib dilutions or solvent control (DMSO). The same lapatinib concentrations were tested in mixture with 1.67 µM, 3.33 or 6.67 µM cisplatin. After α 72-h incubation, adherent cells were ethanol-fixed and epithelial cells were stained using a Cy2™-labeled pan-cytokeratin antibody; then fluorescent colonies were counted. RESULTS: 33 of 51 ex vivo growing HNSCC (64.7%) exhibited epithelial CF allowing for cut-off detection. Lowest cut-off (complete chemotherapeutical suppressed CF) was noted at 6.25 µM lapatinib in three HNSCC (9.1%). The percentage of HNSCC achieving cut-off by 6.67 µM cisplatin (21.2%) was raised by addition of lapatinib at 6.25, 12.5, and 25 µM up to 33%, 45.5%, and 60.6%, respectively. However, we observed significant inter-individual different dose-response curves of HNSCC in response to lapatinib, e.g. variation in concentrations inhibiting CF to 50% (IC(50)) was 60-fold. At the individual level, antagonism, additivity, and synergism were detected as appropriate models describing the mode of action of cisplatin and lapatinib mixtures. CONCLUSION: Lapatinib suppresses CF of epithelial HNSCC cells, and in combination with cisplatin its efficacy is increased. However, caution is advisable due to significant heterogeneity in the response of HNSCC to lapatinib alone and when combined with cisplatin.

Use of Floseal and effects on wound healing and pain in adults undergoing tonsillectomy: randomised comparison versus electrocautery.

The objective of this study was to evaluate the effect of FloSeal(®) (FS, Baxter Healthcare, Deerfield, IL, USA) as a haemostatic matrix in comparison to bipolar electrocautery (EC) after tonsillectomy. Eligible patients were adults undergoing cold-knife tonsillectomy because of recurrent tonsillitis, tonsillar hypertrophy, or peritonsillar abscess (more than 3 months previously). Patients were randomly allocated, on a single-blind basis, to either FS or EC for haemostasis during tonsillectomy. Five experienced surgeons judged the handling of FS application using a five-point scale (very good, good, fair, poor, very poor). Postoperative pain scores were evaluated with a visual analogue scale for 20 days, and the duration under pain medication together with the consumption of pain medication was compared. Wound healing was documented on Days 1-5, 10, and 20. A total of 176 patients were enrolled. Overall, 76/77 (98.7%) of surgeon evaluations of FS handling were judged at least "good". FS-treated patients showed significantly improved wound healing (less thickness of wound plaques) throughout the postoperative observation period, a trend for less postoperative pain (cumulative pain intensity score; P = 0.074), and a significantly shorter duration of pain-medication use (9.5 vs. 11.6 days; P = 0.014) as well as reduced pain-medication consumption/demand (P = 0.032). No difference in the rate of postoperative haemorrhage was observed between the two treatment groups (4.9% for FS patients, 6.0% for EC patients, P = 0.76). In conclusion, this study demonstrates the easy handling of FS application in tonsillectomy. Its use instead of EC after cold-steel tonsillectomy shows beneficial effects on mucosal recovery, as assessed by a decrease in the thickness of wound coating. Furthermore, FS is associated with a significantly shortened duration of pain-medication use and overall reduction in consumption/demand.

The effect of Ginkgo biloba extract (EGb 761) on parameters of oxidative stress in different regions of aging rat brains after acute hypoxia.

BACKGROUND AND AIMS: Neurodegenerative processes of aging seem to be associated with oxidative stress by reactive oxygen species (ROS). This study investigates the influence of age and of acute respiratoric hypoxia on parameters of oxidative stress in different brain regions of Wistar rats and the protective effects of Ginkgo extract (EGb 761) as a radical scavenger. METHODS: Biopsies of frontal and temporal cortices, the cerebellum, and the brainstem of young and old rats (each group n=6-8: normoxic - hypoxic; unprotected - EGb-protected) were analyzed for malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, glutathione (GSH) content, and creatine kinase (CK) activity. Experimental hypoxia: downregulation of oxygen partial pressure to 5 vol. % for 20 minutes. EGb administration: daily 100 mg/kg of body weight in drinking water for 3 months. RESULTS: Effects of age: While most oxidative stress parameters in the temporal cortex, the cerebellum, and the brainstem are increased, this is not the case in the frontal cortex; after additional hypoxia SOD and GSH are diminished in the temporal cortex and the brainstem of old rats. EGb treatment causes contradictory alterations in young, old, and hypoxic brain regions. Minor effects are seen in old hypoxic brains, while there are some protective effects in old normoxic brainstems and cerebellums. CONCLUSIONS: The old brain appears to adapt appropriately to chronic oxidative stress and to the specific conditions of shortterm hypoxia. EGb's protective effect is especially notable in the brainstem and the cerebellum.

Heterogeneity of epithelial and stromal cells of head and neck squamous cell carcinomas in ex vivo chemoresponse.

BACKGROUND: Valid prediction of the effectiveness of chemotherapeutic agents in individual head and neck squamous cell carcinoma (HNSCC) is desirable and might be achieved using ex vivo assays. METHODS: Three biopsies from each of 15 HNSCC were taken, minced and digested by collagenase. The digested HNSCC was added to serial dilutions of either cisplatin (CIS) or docetaxel (DTX), which were prepared under flavin-protecting conditions in ECM-coated microtiterplates. After 72-h incubation, cultures were methanol-fixed and Giemsa-stained. The cutoff concentration (COC; concentration completely suppressing colony formation) for epithelial cells (EC) and stromal cells (SC) was evaluated. RESULTS: 12/15 HNSCC (80%) were evaluable. Despite significant correlation of COC of CIS in respect of colony formation of EC or SC, no significant differences in response of individual HNSCC specimens were found in the t test for paired samples (p > 0.16). The same applied to DTX. However, EC and SC showed heterogeneity in chemoresponses leading to COC variability of more than one titration step in 44.1% (CIS) and 20% of HNSCC (DTX). No significant correlation between the COC of both cell populations was found in HNSCC specimens. CONCLUSIONS: The ex vivo chemoresponse of EC and SC of HNSCC must be analyzed separately.

Nitric oxide synthase (NOS2/3) expression in head and neck squamous cell carcinomas in correlation with clinical patterns.

BACKGROUND: Increased nitric oxide synthase (NOS) expression has been demonstrated in a number of carcinomas and is discussed to play a key role in tumor progression. The aim of this immunohistochemical study was to examine the protein expression rates of endothelial (e)NOS and inducible (i)NOS in head and neck squamous cell carcinomas (HNSCCs) and oral mucosa and to correlate the results with clinicopathologic factors (TN stage). PATIENTS AND METHODS: Protein expression patterns of NOS were studied immunohistochemically (score 0-7) in 58 patients with HNSCC and 7 mucosa samples, and the results were correlated with tumor stages. RESULTS: In oral mucosa, iNOS was only expressed in the basal epithelial layers and in macrophages, eNOS in endothelial cells and lymphocytes. In contrast, both NOS isoforms were expressed in HNSCC with preference at the tumor margins. 64% of tumor specimens demonstrated a positive eNOS immunoreactivity (score > or =3), 55% a positive iNOS immunoreactivity. NOS protein expression rates reached higher scores in tumors of patients with lymph node metastasis (N > 0; iNOS protein expression rate p < 0.05). CONCLUSIONS: HNSCCs are able to express both NOS protein isoforms in relevant amounts, and we presume that synthesized NO is able to support angiogenetic patterns and facilitate tumor progression and lymphatic spread.

Single tissue samples from head and neck squamous cell carcinomas are representative regarding the entire tumor's chemosensitivity to cisplatin and docetaxel.

BACKGROUND: In multimodal therapy concepts for advanced head and neck squamous cell carcinoma (HNSCC), a valid predictive assay for the quick detection of efficient chemotherapeutic agents is desirable. Questionable so far was whether tissue samples of about 100 mg correctly reflect the chemoresponse of a whole HNSCC. This was proven using an ex-vivo colony-forming assay. MATERIALS AND METHODS: Of 14 HNSCC, 3 biopsies each were taken from separate sites, minced, and collagenase digested. HNSCC digests were added to microtiter plates containing serial dilutions of chemotherapeutic agents or medium as control. After 72-h incubation, wells were washed and cultures methanol-fixed before Giemsa-staining. Epithelial colonies were counted. RESULTS: 11/14 HNSCC (78.6%) showed sufficient colony formation allowing reliable cut-off detection. Cut-off concentrations (complete chemotherapeutically suppressed colony formation) between 3.3 microM and >50 microM cisplatin, and 0.55 microM and 17.6 microM docetaxel were detected. Inhibition of colony formation to 50% of colonies detected in controls (IC50) was found between 0.2 microM and 17.9 microM cisplatin or 1.5 microM and 13.7 microM docetaxel. Cut-off concentrations and IC50 of the HNSCC fragments showed a strong correlation (docetaxel: r > 0.80, p < 0.005; cisplatin: r > 0.67, p < 0.044), while being only insignificantly different in the t-test for paired samples (docetaxel: p > 0.163; cisplatin: p > 0.167). CONCLUSION: In most cases, tissue samples of about 100 mg allow a representative assessment of chemoresponse of HNSCC.

Cardioprotective effect of EGb 761 on myocardial ultrastructure of young and old rat heart and antioxidant status during acute hypoxia.

BACKGROUND AND AIMS: Acute hypoxia is a threatening clinical case of emergency and may result in ultrastructural damage, with complete loss of cellular and organ functions. However, little is known about the differences in hypoxia tolerance between young and old myocardia and the protective effects of radical scavenging agents in acute hypoxic stress situations. METHODS: We investigated the age-dependent differences of the myocardial ultrastructure and antioxidative status (superoxide-dismutase (SOD) activity and malondialdehyde (MDA) content) of young (6 months) and old (22-24 months) Wistar rats (Crl (Wi)Br) after acute respiratory hypoxia of 20 min at 5% v/v O2 in N2O mixture, and the protective effect of Ginkgo biloba extract (EGb 761). RESULTS: Ultrastructural-morphometric and biochemical age analysis only revealed a decrease in the sarcoplasma volume fraction, an increase in homogeneous intramitochondrial areas, significant higher SOD activity and lower MDA levels in the group of old rats. Pretreatment with EGb 761 led to a significant decrease in MDA content in both control groups. Acute hypoxic stress increased the volume fractions of sarcoplasmatic reticulum, t-tubules, vacuoles, and lipid droplets, and caused mitochondrial swelling, with a more significant increase in degenerated and homogeneous intramitochondrial areas in the old group. SOD activity decreased only in the old hypoxic group; MDA content fell in both. Pretreatment with EGb 761 reduced ultrastructural-morphometric hypoxic damage in both groups, significantly below the levels of control. Young rat myocardium showed significantly higher SOD activity after hypoxia than untreated or older specimens. CONCLUSIONS: Better hypoxia tolerance is demonstrated by the young myocardium, and an obvious hypoxia-protective effect of EGb 761 in both age groups.