Identification of biomarkers for apoptosis in cancer cell lines using metabolomics: tools for individualized medicine.

BACKGROUND: Metabolomics is a versatile unbiased method to search for biomarkers of human disease. In particular, one approach in cancer therapy is to promote apoptosis in tumour cells; this could be improved with specific biomarkers of apoptosis for monitoring treatment. We recently observed specific metabolic patterns in apoptotic cell lines; however, in that study, apoptosis was only induced with one pro-apoptotic agent, staurosporine. OBJECTIVE: The aim of this study was to find novel biomarkers of apoptosis by verifying our previous findings using two further pro-apoptotic agents, 5-fluorouracil and etoposide, that are commonly used in anticancer treatment. METHODS: Metabolic parameters were assessed in HepG2 and HEK293 cells using the newborn screening assay adapted for cell culture approaches, quantifying the levels of amino acids and acylcarnitines with mass spectrometry. RESULTS: We were able to identify apoptosis-specific changes in the metabolite profile. Moreover, the amino acids alanine and glutamate were both significantly up-regulated in apoptotic HepG2 and HEK293 cells irrespective of the apoptosis inducer. CONCLUSION: Our observations clearly indicate the potential of metabolomics in detecting metabolic biomarkers applicable in theranostics and for monitoring drug efficacy.

Metastasis of a renal cell carcinoma to the nose and paranasal sinuses.

We present a case of a metastasis of a renal cell carcinoma to the nose and paranasal sinuses. A 66 year old male patient developed a mass in his left nasal cavity and paranasal sinuses, five years after he underwent a left sided nefrectomy for a renal cell carcinoma. The histopathologic examination of the nasal mass showed metastasis of a renal cell carcinoma. A craniofacial resection was performed followed by radiotherapy.

Contractile action of galanin analogues on rat isolated gastric fundus strips is modified by tachyphylaxis to substance P.

This study was undertaken to characterize the interaction of porcine galanin (Gal) and some of its analogues with their receptors on rat gastric fundus muscle strips. Gal, galantide (M15) and Gal(1-14)-[Abu8]SCY-I evoked concentration-dependent contractions of gastric smooth muscle strips. Reproducible effects were observed in concentrations of 1-300, 3-1000 and 100-3000 nM, respectively. Specific EC50 for the contractile effect equalled 13.70 and 187 nM. Hill's coefficient for Gal is 1.03 indicating an interaction of one Gal molecule with one receptor, fulfilling the criteria of classical receptor theory. For M15 and Gal(1-14)-[Abu8]SCY-I Hill's coefficients are different from 1, namely 0.73 and 1.56, pointing out that the principle of interaction of one drug molecule with one receptor may not apply. The contraction induced by 300 nM of Gal was not significantly modified by tachyphylaxis to substance P (SP). On the contrary the introduction of tachyphylaxis to SP decreased the contractile effects of M15 and Gal(1-14)-[Abu8]SCY-I by about 57.7 +/- 3% and 39.6 +/- 5%, respectively. The findings suggest that contractile actions of M15 and Gal(1-14)-[Abu8]SCY-I are probably not only due to their agonist activities at Gal receptors but may result from a subsequent stimulation of receptors for SP or release of endogenous SP.

Tumour-associated tissue eosinophilia in head and neck squamous-cell carcinoma.

The possible role of tumour-associated tissue eosinophilia (TATE) as a prognostic factor in epithelial tumours has been investigated before. TATE is supposed to be associated with good prognosis. In this study concerning 63 patients with laryngeal and/or pharyngeal squamous-cell carcinoma, we tried to verify this hypothesis. Of the 25 patients with TATE, 17 (68%) had a good outcome opposed to 8 (32%) with a poor outcome. This numerically better chance for a good outcome in case of TATE was however not statistically significant. Uniformity of research material, staining and classification of TATE together with more representative and larger series of patients should ascertain the real value of TATE as a valid prognostic factor in head and neck squamous-cell carcinoma.

Desensitisation of substance P decreases the contractile effect of M15 and Gal (1-14)-(Abu 8) SCY-I on the rat gastric fundus.

Tachyphylaxis to SP decreased the effect of M15 and Gal(1-14)-[Abu8]SCY-I on gastric smooth muscles, without effect on the action of Gal. These findings support our initial hypothesis: the action of M15 and Gal(1-14)-[Abu8]SCY-I on the smooth muscles may not only be due to their agonist activity at Gal receptors, but may result from a subsequent stimulation of receptors for SP and perhaps for other tachykinins as well, however, a possibility that Gal analogues release endogenous SP can not be excluded. Further studies involving a tachykinin antagonist (spantide) are in progress at the moment. [1].

Actions of galanin and some of its analogues on rat isolated gastric fundus.

The study was undertaken to characterize the effects of porcine galanin (pGal) and some of its analogues on rat gastric fundus muscle strips. pGal, galantide (M15) and pGal(1-14)-[Abu8]SCY-I evoked reproducible concentration-dependent contractions in concentrations of 1-300, 3-1,000 and 100-3,000 nM, respectively, with EC50 values of 13, 70 and 187 nM. Hill's coefficient for pGal is 1.03, indicating an interaction of one pGal molecule with one receptor, fulfilling criteria of classical receptor theory. For M15 and pGal(1-14)-[Abu8]SCY-I, Hill's coefficients are significantly different from 1, namely 0.73 and 1.56, so that one drug molecule may not interact with one receptor. The stimulatory effects of pGal were not modified by dibenamine 10 microM or glybenclamide 1 or 10 microM. Diltiazem 0.1, 1 and 10 microM, papaverine 0.1, 10 microM or dibutyryl cAMP (dib cAMP) 100 and 300 microM, blocked the contraction to pGal in a concentration-dependent manner, indicating an important role for the influx of extracellular calcium ions and regulation by cAMP the pGal-evoked contraction. Diltiazem, dibutyryl cAMP and papaverine were not competitive antagonists of pGal in the stomach smooth muscle.

Clinical approach to the dysphagic patient.

Dysphagia is a symptom of disease reflecting either structural or neuromuscular disorders of the oropharynx or esophagus. Dysphagia should be distinguished from globus sensation and odynophagia. The clinical evaluation of the dysphagia patient by the otolaryngologist requires taking a detailed history of the complaint, extended ENT physical examination, fiberoptic nasopharyngolaryngoscopy, to assess the oral and pharyngeal phases of swallowing. Special investigations including radiography, esophageal endoscopy, ultrasonography, pH metry, manometry may be needed for the full diagnosis. It is essential for the otolaryngologist to develop a close working relationship with the radiologist, gastroenterologist, neurologist and swallowing therapist. This team approach will allow in most of the patients with dysphagia not only to identify the cause of the complaint but also to implement an effective treatment.

Surgical treatment of oropharyngeal swallowing disorders.

The objectives of the surgical treatment of oro-pharyngeal swallowing disorders is (1) to improve the impaired transport of the bolus, and (2) to prevent aspiration or control it to safe levels. In this overview the common surgical procedures to improve the specific oro-pharyngeal swallowing disorders are presented. The indications, advantages and disadvantages of each surgical technique are discussed.

Fluctuating hearing loss in sarcoidosis.

Sarcoidosis is a systemic granulomatous disease, which can affect the vestibulo-acoustic system at any level, causing vertigo and/or hearing loss. The main mechanism could be vasculitis. One patient with fluctuating hearing loss as the first manifestation of a systemic sarcoidosis is presented. Diagnostic modalities, especially different sites of biopsy or needle aspiration are discussed. The treatment is steroid therapy. Hearing will be restored provided no irreversible changes of the VIIIth nerve and/or the inner ear have occurred.

Ototoxicity of an ototopic preparation--experimental results and clinical facts.

The ototoxicity of "Cortisporin Otic Suspension" containing 3.5 ml neomycin base, 10,000 units/ml polymyxin B was studied using the baboon (papio ursinus) as an experimental animal. Inner and outer hair cell loss occurred in all cochleas of the ears which received cortisporin. Most lesions were confined to the basal turn of the organ of Corti. Separate evaluation of polymyxin B and neomycin at the same concentration as in cortisporin showed that polymyxin B produced greater cochlear damage. The results of this study and clinical reports of sensorineural hearing loss due to commercially available ear drops indicate that ototopical preparations must be used with caution in clinical situations in which they may reach the middle ear.

[Premalignant lesions of the laryngeal epithelium]. / Premaligne lesies van het larynxepitheel.

At the Academic Hospital of the Free University of Brussels 243 patients underwent microlaryngoscopy over a period of 11 years (1978-1989). Histologic examination disclosed premalignant lesions in 55 patients. 26 patients had a follow-up (mean follow-up time was 5 years) and were selected for this study. The patients with premalignant lesions were divided into 3 groups: group 1 = hyperplasia and/or keratosis with or without mild dysplasia (11 patients); group 2 = mild dysplasia (7 patients); group 3 = severe dysplasia and carcinoma in situ (8 patients). In group 1 no malignant transformation was seen. In group 2 one invasive carcinoma developed after a period of 5 years (14%) and in group 3 two malignant transformations appeared 14 and 15 months after the initial diagnosis (25%). The treatment of choice was a total excision biopsy or stripping. A life time follow-up with an interval period of 6 months is recommended. The patients were also motivated to change their smoking habits.

[Laryngeal amyloidosis]. / Laryngeale amyloidose.

Laryngeal amyloidosis is a rare, benign disease of the larynx. An update on the pathology and clinical implications of this affection, along with the actual treatment modalities are discussed.

Density of epithelial cells in the normal human nose and the paranasal sinus mucosa. A scanning electron microscopic study.

214 biopsies of mucosa from various sites of the nose and paranasal sinuses were obtained post-mortem and examined using the scanning electron microscope. The density of ciliated cells was increased in the nasal cavity, in the antero-posterior direction. In paranasal sinuses the density of these cells was high, except near the ostium of the maxillary sinus, where the density was decreased by half. Non-ciliated epithelial cells were found in relatively few areas: anterior aspect of the middle and lower turbinates, anterior septum. However, no uniform distribution of these cells was present. The density of goblet cells was significantly lower in the paranasal sinuses as compared to the nasal cavity, with the highest density being found near the ostium of the maxillary sinus.

Ototoxicity of neomycin and polymyxin B following middle ear application in the chinchilla and baboon.

Previous experimental studies have demonstrated structural damage of the organ of Corti and stria vascularis following application of combination antibiotic otic drops to the middle ear. In this investigation the ototoxic effects of neomycin and polymyxin B (two antibiotics often used together in ototopical preparations) were separately evaluated after administration of each agent to the middle ear cavities of chinchillas and baboons. The antibiotics were administered in saline solution at the same concentrations used in Cortisporin Otic Suspension (3.5 mg/ml neomycin base, 10,000 units/ml polymyxin B). In both the rodent and primate, polymyxin B consistently produced greater cochlear damage than did neomycin. In fact, the extent of hair cell loss and strial injury produced by polymyxin B alone was, in many cases, comparable to that previously observed after application of Cortisporin Otic Suspension itself. Hair cell loss in the baboon was markedly less severe than in the chinchilla. It is believed that differences in position and structure of the round window membrane are important factors in the differing levels of ototoxicity observed in the rodent and primate.

Prevalence of otitis media in a Venda village.

During mid-winter in 1985, 263 children and 217 adults from a remote rural community in Venda, were examined for the presence of middle ear lesions by: pneumatic otoscopic examination; tympanometry; and contralateral acoustic reflex. A diagnostic algorithm was applied to assess the findings. The prevalence of otitis media (OM) in children (less than or equal to 15 years) was 8.4% and in adults (greater than 15 years) 2.3%, with the highest figure in the 0-23 months age group. OM with effusion was found in 3.6% of the children. Our data suggest the existence of considerable differences in the prevalence of OM between comparable black rural communities.

Ototoxicity of an ototopical preparation in a primate.

Severe ototoxicity has been observed in laboratory rodents after middle ear application of ototopical preparations or their constituents. Owing to their relatively high susceptibility, these animals may not be ideal experimental models. In the present study the ototoxicity of Cortisporin Otic Suspension (neomycin, polymyxin B, hydrocortisone) was studied using the baboon as a primate model. Middle and inner ear pathologic changes were assessed after a single application of the suspension to the tympanic cavity. Inner and outer hair cell loss occurred in all the experimental animals; it was, however, confined to the basal turn of the organ of Corti. Relative to previously studied rodents, the thicker, more densely structured round window membrane of the primate appears to provide better protection against penetration of ototoxic agents. Nonetheless, the results of this study indicate that ototopical preparations must be used with caution in clinical situations in which they may reach the middle ear.

Prevalence of otitis media in children in a black rural community in Venda (South Africa).

During mid-winter in 1985, 480 residents from a remote rural community in Venda, South Africa, were examined for the presence of middle and external ear pathology. The children (less than 15 years) numbered 267. The otological examination consisted of (1) pneumatic otoscopic examination, (2) tympanometry, (3) contralateral acoustic reflex. A diagnostic algorithm as proposed by Cantekin et al. was applied to assess the findings. Our data showed the prevalence of otitis media in children to be 8.2%. The highest prevalence was found in the 0-23 months age group. Of 22 children with otitis media, only one had a perforation with otorrhea. Otitis media with effusion was found in 3.8% of the children. External otitis media as an independent variable was found in only 1.5% of the 267 children, while 6.7% presented with scarred tympanic membranes. Our data suggest the existence of considerable differences in the prevalence of otitis media in children between two comparable black rural communities from Nigeria and Venda.