Absence of high-risk human papilloma virus in p16 positive inverted sinonasal papilloma.

OBJECTIVES: Sinonasal inverted papilloma (SIP) is a relatively rare disease, and its etiology is not understood. It is characterized by locally aggressive growth and a strong tendency to recur despite its benign histology. AIMS: The aim of this study was to identify the presence of human papilloma virus (HPV) and its surrogate marker p16 in SIP tissue samples from a regional cohort. MATERIAL AND METHODS: Subjects were identified from our regional center cohort of 88 SIP patients treated between 1984-2014. From these subjects, 54 were included in this study. Of these, 53 biopsies were analyzed with PCR, and 54 samples were immunohistochemically stained for p16. DNA was extracted from histopathologically verified SIP. Genotype screening for 13 high risk-, 5 oncogenic and 6 low risk HPV types was performed using the PapilloCheck® HPV-screening test. RESULTS: HPV analysis was successful for 38 of 53 samples. Of the 38 successfully analyzed samples, only 2 samples were positive for HPV 11. Notably, p16 was present in the epithelia in all samples, and in the papilloma lesions in 37 samples. CONCLUSION: Since only 2 out of 38 SIPs were positive for HPV (type 11), and at the same time p16 was positive in epithelia in all samples and in 37 of 38 papilloma lesions of the samples, it is concluded that p16 cannot be used as a surrogate marker for high-risk HPV-infection in SIP. We are currently planning a prospective, multicenter study in order to increase the study power and in order to be able to better evaluate the clinical implications of HPV-and p16 in SIP.

Pharmacological intervention in the field of ototoxicity.

Modern research on ototoxicity goes back to the 1940s, when streptomycin was introduced into clinical practice. Today, aminoglycoside antibiotics and platinum-based chemotherapy, mainly cisplatin, are the most important drugs that damage the inner ear and cause hearing loss. The mode of drug administration as well as drug characteristics influence the likelihood that adequate monitoring of drug pharmacokinetics can be performed. It is not possible to predict the individual risk of treatment with an ototoxic drug, but identification of high-risk treatment protocols is important. There are many studies ongoing with the aim of discovering and developing drugs to treat different types of inner ear disorders. The mechanisms of ototoxicity and subsequent loss of hearing function have been mapped in various experimental models and have provided us with useful information for developing protective treatment. When an ototoxic lesion is established, restoration of hearing function becomes more difficult. For both aminoglycoside antibiotics and cisplatin, a large number of otoprotectors have been suggested. Systemic co-administration of an otoprotector would be the easiest approach to avoid ototoxicity in patients but it may negatively affect the intended pharmacotherapeutic aim of the ototoxic drug. New pharmacological formulations are being developed for local otoprotective treatment. This short review focuses on results from clinical reports on otoprotection in patients treated with aminoglycoside antibiotics and cisplatin. So far there is limited evidence for the safe management of otoprotection in patients. Further high-quality studies are needed to provide reliable data on the safety and effectiveness of pharmacological interventions to reduce drug-induced hearing loss.

Expression of p16 in squamous cell carcinoma of the mobile tongue is independent of HPV infection despite presence of the HPV-receptor syndecan-1.

BACKGROUND: Tongue squamous cell carcinoma (TSCC) is increasing in incidence, especially among young patients and preferably females. Infection with human papilloma virus (HPV) has been suggested as a cause of SCC in the head and neck, and the proportion of oropharyngeal cancers caused by HPV has steadily increased. METHODS: Samples from 109 patients with primary TSCC were analysed for the presence of HPV16 by in situ hybridisation and for expression of its surrogate marker p16 and the HPV receptor syndecan-1 by immunhistochemistry. RESULTS: No evidence of HPV16 DNA was observed in the tumours, although one-third showed p16 staining. There was no difference in the expression of the primary HPV receptor, syndecan-1, between TSCC and a group of tonsil SCC. CONCLUSION: Whereas p16 is expressed in some TSCCs, HPV16 is undetectable, therefore, p16 cannot be used as a surrogate marker for high-risk HPV-infection in this tumour. Despite presence of the HPV-receptor syndecan-1 in TSCC, HPV prefers the tonsillar environment. Lack of p16 associates with worse prognosis primarily in patients aged ⩽40 years with tongue SCC. The improved prognosis seen in p16-positive TSCC can be due to induction of a senescent phenotype or an inherent radiosensitivity due to the ability of p16 to inhibit homologous recombination repair.

Downregulation of miRNA-424: a sign of field cancerisation in clinically normal tongue adjacent to squamous cell carcinoma.

BACKGROUND: The overall survival for patients with squamous cell carcinoma of the tongue is low and the search for early diagnostic and prognostic markers is thus essential. MicroRNAs have been suggested as potential prognostic and diagnostic candidates in squamous cell carcinoma of head and neck in general. METHODS: On the basis of the known differences between sub-sites within the oral cavity, we investigated the expression and role of microRNA-424 in squamous cell carcinoma arising in tongue. MicroRNA levels were measured by qRT-PCR in both tissue and plasma samples. RESULTS: Levels of microRNA-424 were upregulated in tongue squamous cell carcinoma, but not in tumours originating from gingiva or floor of the mouth. Interestingly, microRNA-424 was downregulated in clinically normal tongue tissue next to tumour compared with completely healthy tongue, indicating that microRNA-424 could be a marker of field cancerisation in this tumour type. However, expression of microRNA-424 in a tongue-derived epithelial cell line revealed no significant changes in the expression profile of proteins and genes. CONCLUSIONS: Our patient data show that microRNA-424 alterations are a marker of field cancerisation specific for tongue tumourigenesis, which also could have a role in development of tongue squamous cell carcinoma.

miRNA analysis of formalin-fixed squamous cell carcinomas of the tongue is affected by age of the samples.

Global miRNA expression arrays were used for analysis of 836 miRNAs in formalin-fixed paraffin-embedded samples from 21 tongue cancer patients and 8 controls. Samples had been stored for one to eleven years. Results separated tumour samples from controls, however, the largest variation was correlated to sample storage time, detectable already after one year. With the use of a linear regression model we could adjust for the storage-dependent effect, leading to the identification of 54 differentially expressed miRNAs in tongue cancer, compared to 16 when using standard normalization, including up-regulation of a novel miRNA, miR-424.

Percutaneous endoscopic gastrostomy (PEG) - a long-term follow-up study in head and neck cancer patients.

Many patients with head and neck cancer experience problems related to swallowing. A retrospective study of 156 consecutive patients who received a percutaneous endoscopic gastrostomy (PEG) at a teaching hospital is presented. The results showed that 42% had complications. Fatal complications were seen in connection with PEG tube placement, but severe and minor complications could occur much later. The method of PEG tube insertion did not affect the complication rates. The spectrum of observed complications is different to that reported earlier, suggesting that the learning curve of surgeons under training could have influenced the outcome. It may be concluded that for a very sick patient a theoretically easy surgical procedure could turn into a potentially dangerous operation. It is important to select suitable candidates for a PEG. Head and neck cancer patients with a PEG need special attention in connection with the PEG tube placement and also in a long perspective, e.g. by follow-up at a nurse-led outpatient clinic.

Intracochlear administration of thiourea protects against cisplatin-induced outer hair cell loss in the guinea pig.

Amelioration of cisplatin-induced side-effects is of great clinical importance. Local administration of a cytoprotective agent to the inner ear offers a possibility to prevent cisplatin-induced ototoxicity without risk of interference with the antitumour effect. The ideal substance for local administration has yet to be identified. Thiourea (TU) has unique properties that make it an interesting candidate. This study was initiated to test the hypothesis that TU given by local administration protects against cisplatin ototoxicity in the guinea pig. After baseline auditory brainstem response (ABR) assessment, the left cochlea was implanted with a microtip catheter connected to an osmotic pump filled with either 27 mg/ml TU in artificial perilymph (AP), or AP administered for the full duration of the study. Three days post-implant, animals with normal ABRs received an intravenous injection of 8 mg/kg body-weight cisplatin. Five days after the cisplatin treatment ABRs were reassessed, animals decapitated and bilateral cytocochleograms prepared. TU-treated ears demonstrated significantly lower outer hair cell (OHC) loss as compared to contralateral untreated ears, and significantly lower OHC loss compared to AP-treated ears. ABR threshold shift did not differ significantly between the two groups. It can be postulated that TU demonstrates partial protection against cisplatin-induced ototoxicity.

Cisplatin-induced hearing loss: influence of the mode of drug administration in the guinea pig.

Cisplatin (8 mg/kg) was given intravenously to guinea pigs either as a 15 s bolus injection (25 animals) or as a 1 h infusion (28 animals). To determine the influence of the mode of cisplatin administration and pharmacokinetics on the ototoxic side-effect, the concentrations of cisplatin and the biotransformation product monoaquated cisplatin were determined in blood ultrafiltrate using liquid chromatography with post-column derivatization. Ototoxic effect was evaluated as difference in pre- and 96 h post-exposure auditory brainstem response (ABR) threshold. The cisplatin peak concentration was considerably higher, 19.2+/-2.4 microg/ml, in the bolus injection group than in the infusion group, 6.7+/-0.5 microg/ml (mean+/-S.E.M.). The area under the blood ultrafiltrate concentration time curve (AUC) for cisplatin was slightly greater in the infusion group, 442+/-26 microg/ml/min, than in the bolus injection group, 340+/-5 microg/ml/min. For monoaqua cisplatin, the AUC was not different between the groups (bolus injection: 30.8+/-1. 5 microg/ml/min, infusion: 34.1+/-3.3 microg/ml/min). A significant ototoxic effect was observed in both groups at 20 and 12.5 kHz, but there was no difference between the groups in the extent of threshold shift. The interindividual variability in susceptibility to ABR threshold shift was far greater than the variability in pharmacokinetics, suggesting that other factors are more important in determining the degree of hearing loss.

Blood-perilymph barrier and ototoxicity: an in vivo study in the rat.

Cisplatin and gentamicin are two ototoxicants that are supposed to be transported by the paracellar route, i.e. via cellular junctions, to the perilymphatic compartment. This study was initiated to test the hypothesis that susceptive variation of individuals to ototoxic drugs may be explained by variability in transport properties. The transport of radioactive mannitol through the blood-perilymph barrier was correlated in vivo with the acute effect of cisplatin and gentamicin on auditory function. Transport of radioactive mannitol across the blood-perilymph barrier was monitored by sampling of scala vestibuli perilymph at 60 and 120 min after an intravenous infusion of the tracer to nephrectomized Long Evans rats. Counting of hair cell loss was performed in the animals receiving 16 mg/kg body weight cisplatin. The transport of radioactive mannitol across the blood-perilymph barrier did not correlate with the ototoxic effect of cisplatin, evaluated as changes in the auditory-evoked brainstem response thresholds or loss of outer hair cells. The results provide evidence that the barrier function is not involved in the interindividual variability of the ototoxic effect of cisplatin. Furthermore, it can be postulated that neither cisplatin nor gentamicin induce a disruption of endothelial cell junction stability in the inner ear.

Field screening of blood lead levels in remote Andean villages.

Blood lead (PbB) levels were investigated in chronically lead (Pb) exposed Andean children and adults living in a highly Pb contaminated area of Ecuador where Pb glazing of ceramics is prevalent. A comparative study was made of the PbB levels of Pb-glazing and non-Pb-glazing families living in close proximity, using three PbB analysis techniques. Fifty-one, 50-microl blood samples from children and adults were analyzed in the field by a finger-stick capillary screening technique using the portable ESA LeadCare Blood Lead Testing System (LCS). Venous blood samples of 2-4 ml were collected from the same 51 participants and analyzed in the laboratory by inductively coupled plasma mass spectrometry (ICP-MS) and by graphite furnace atomic absorption spectrometry (AAS). The median PbB levels for the Pb-glazing group as determined by the ICP-MS, AAS and LCS techniques were 37.2 microg/dl (range 11.6-101.0), 32.0 microg/dl (range 8.0-70.0 microg/dl) and 44.0 microg/dl (range 19.0-105.0), respectively. The median PbB levels for the non-Pb-glazing group were 9.2 microg/dl (range 5.0-21.7) with ICP-MS, 9.0 microg/dl (range 4.3-32.0) with AAS, and 11.3 microg/dl (range 7.3-21.1) with LCS. The differences in PbB levels between the Pb glazing and non-Pb glazing groups were statistically significant (p = < .0001) for each PbB analysis method. Correlations between paired samples were: LCS and ICP-MS: r = 0.913, LCS and AAS: r = 0.829, and ICP-MS and AAS: r = 0.905. The results suggest that neighboring Pb glazing and non-Pb glazing families have significantly different PbB levels, and that the portable LCS field technique may be useful for screening and periodic monitoring of relatively low and high PbB levels of persons in remote high altitude Andean areas.

Paracellular transport properties of inner ear barriers do not account for cisplatin toxicity in the rat.

The interindividual variability for the ototoxic effect of the antineoplastic drug cisplatin has still to be explained. To examine if the variability can be related to differences in drug kinetics, the effect of cisplatin on the paracellular transport properties of the inner ear barriers was studied in vivo in cisplatin treated Long-Evans rats. The concentration of [3H]mannitol was followed in plasma, scala vestibuli perilymph, and endolymph after an intravenous infusion of the tracer. Cisplatin had no effect on paracellular transport of the inner ear barriers 3 days after administration of 8 mg/kg cisplatin. However, an interindividual variability for the transport of [3H]mannitol across the blood-perilymph barrier was evident, indicating a variability for the passive transport of solutes to the inner ear.

Blood flow-independent accumulation of cisplatin in the guinea pig cochlea.

Considerable interindividual variability in the ototoxic effect of cisplatin has become the unpredictable dose-limiting factor in its use as curative as well as palliative therapy. The drug accumulates in highly vascular areas in the cochlea, causing dose-related hair cell loss. The purpose of this study was to assess blood flow-dependent aspects of cisplatin absorption in the cochlea in order to better understand factors that may influence cisplatin-induced ototoxicity. The effect of reduced cochlear blood flow on the ototoxic action of cisplatin was studied in guinea pigs. Before cisplatin administration the cochlear vasculature in each animal was unilaterally pre-constricted, by the application of 2% epinephrine to the round window. A 20-30% reduction in cochlear blood flow, assessed by laser Doppler flowmetry, was maintained before and after intravenous infusion of 0.1% cisplatin. Cisplatin infusion affected cochlear blood flow but not vessel conductivity. The cochlear blood flow decrease, maintained by local epinephrine application to the round window during cisplatin infusion, did not alter the cisplatin-induced hearing loss. In addition, the concentration of free cisplatin in scala tympani perilymph did not differ between epinephrine-treated and non-treated ears. Our results indicate that cisplatin transport into the cochlea is not an energy-dependent process in the lateral wall vasculature.

Radiotherapy enhanced ototoxicity of cisplatin in children.

We reviewed and re-examined 31 children (6 months-14 years at the time of diagnosis), who had been treated for a neoplasm in Turku University Central Hospital between 1989 and 1994. The children were divided into 3 groups according to the site of the neoplasm and the type of therapy. Group I included 13 children, operated on for an intracranial tumor and received postoperative radio- and cisplatin-based chemotherapy. Group II included 14 children operated on for intracranial tumors and treated with radiotherapy, but not given chemotherapy. Group III included 4 children suffering from extracranial malignancies and they had received chemotherapy including cisplatin. The children in Group I had significantly worse hearing thresholds in the middle- and high-frequency range than children in Groups II and III. In a precise analysis of the different factors, no single dose of cisplatin, inner ear irradiation dose or totally to the central nervous system (CNS) received irradiation dose correlated to the detected hearing loss. However, multiple linear correlation analyses suggest a combined effect of radiotherapy plus cisplatin resulting in a high frequency hearing loss. This is in accordance with earlier random case reports, and supports the idea that radiotherapy should be considered cautiously in children treated with cisplatin for intracranial malignancies.

Cisplatin administration to gynecologic cancer patients. Long-term effects on hearing.

BACKGROUND: Cisplatin is known to create an acute dose-related ototoxic effect. There are unanswered questions regarding the long term effect of cisplatin on hearing in gynecologic cancer patients. METHODS: A retrospective review of 59 to 115 months' duration was performed on 184 women with gynecologic cancer who were treated with cisplatin-based chemotherapy between 1982 and 1986. Twenty-six of 40 survivors were again tested audiometrically with the same audiologic equipment. RESULTS: Fourteen patients (54%) had significantly progressive hearing loss (> or = 15 decibels) at long term follow-up compared with the posttreatment control. These changes were generally small and restricted to three frequencies or fewer in one of the patient's ears. The changes corresponded to the expected age effect upon hearing. Only 2 patients (8%) showed more severe hearing threshold changes. The hearing loss in one of the two patients might represent degenerative changes induced by cisplatin treatment, whereas in the other patient the etiologic background to the hearing loss remains unknown. CONCLUSIONS: This study does not provide any strong evidence for a delayed ototoxic effect of cisplatin that should influence therapeutic strategy. Patients who receive moderate dose cisplatin therapy, 50 mg/m2 per body surface area every 4 weeks, have a negligible long term risk of a drug-induced social hearing handicap.

Comparative entry of carboplatin and sucrose in endolymph in the rat cochlea.

The permeability of the perilymphatic-endolymphatic interface for carboplatin was determined after lateral cerebral ventricle infusion of radioactive carboplatin (cis-diamine[1,1-cyclobutane-1-14C-dicarboxylate]platinum) to rats. [14C]sucrose, a similar weight molecule was used for comparison of the kinetics in the inner ear fluids. 14C-radioactivity was measured in perilymph and endolymph. The rate of elimination of the tracers from perilymph was equivalent indicating no difference in transport across the blood-perilymph barrier. The transport from perilymph to endolymph was very restricted for both substances. The present study indicates that the ototoxic effect of carboplatin cannot be explained by a specific endolymphatic transport mechanism.

Effect of cisplatin administration on the electrochemical composition of endolymph in the rat cochlea.

The effect of cisplatin on the electrochemical composition of the cochlear endolymph was studied in Long-Evans rats three days after a single intraperitoneal injection (8 mg/kg b.w.). A dose 2/3 of LD50 induced a decrease of the endolymphatic concentration of potassium whereas the endocochlear potential was unaffected. The discrepancy between these two findings indicated that cisplatin did not alter the mechanisms involved in the genesis of the endocochlear potential but modified the passive K transport into endolymph.

Distribution of cisplatin in perilymph and cerebrospinal fluid after intravenous administration in the guinea pig.

The concentration of free cisplatin was followed in plasma, scala tympani perilymph and cerebrospinal fluid (CSF) after an intravenous injection (12.5 mg/kg) in guinea pigs. Liquid chromatography with postcolumn derivatization was used for quantitative determination of the drug. The distribution of cisplatin to CSF was fast; at 10 min after drug administration the concentration was 7 micrograms/ml and the CSF:plasma ratio was 0.37. Cisplatin seems to distribute more slowly to the perilymphatic compartment. The highest concentration measured was 4 micrograms/ml at 20 min after the injection, and the perilymph:plasma ratio was 0.40 at that time. The concentration-time curves generated for cisplatin in perilymph and CSF were similar. No accumulation in the perilymphatic compartment or CSF was observed.

Monitoring of hearing during treatment of leukaemia with special reference to the use of amikacin.

Thirty-nine patients with leukaemia were followed audiometrically during treatment with broad-spectrum antibiotics. Amikacin was given during neutropenic febrile episodes. Five patients reported a deterioration of the hearing function after termination of amikacin treatment. Significant hearing threshold loss occurred in 20 patients (51%). The hearing threshold changes were small in general, except for two patients who exhibited bilateral hearing threshold changes in the frequency range 0.5-8 kHz. Using multiple linear regression analysis 22% of the changes in hearing thresholds was estimated to be related to old age, an increased trough concentration of amikacin and an impaired pretreatment hearing state. Factors found not to influence the hearing thresholds were maximum peak concentration of amikacin, cumulative duration of therapy, pretreatment renal dysfunction and concomitant use of vancomycin. It is concluded that administration of amikacin for repeated treatment courses is associated with a low incidence of serious changes in hearing function.

The relative importance of the routes and sources of wound contamination during general surgery. II. Airborne.

The influence of airborne bacteria on wound contamination during biliary surgery was studied. When bacteria grew in the bile they accounted for most of the bacteria in the wound but when the wounds were free of bile bacteria many of the bacteria came from the patient's skin. It was only in wounds with little contamination from non-airborne routes that it was possible to demonstrate an effect of airborne contamination. In such a situation it was estimated that a reduction in the airborne bacteria in the operating room of about 13-fold would reduce the wound contamination by about 50%. The contamination of patient drapes from various sources and its relationship to wound contamination was studied. It was demonstrated that in areas away from the wound, the bacterial concentration on the drape surface was significantly affected only by airborne bacteria. In the area close to the wound, airborne bacteria and bacteria from the wound significantly affected drape contamination. However, it was found that more bacteria transferred from the wound to the drape surface than vice versa. Punctured gloves, impervious gowns and the number of bacteria on the patient's skin did not significantly affect the counts on the drapes' surfaces.