Plasmatic cytokine network in patients with laryngeal carcinoma after surgical treatment.

Alterations in host immunity, inflammation, angiogenesis and metabolism are all prominent clinical features in patients with laryngeal squamous cell carcinoma (LSCC). Although the origin of the signals and mechanisms underlying these responses are not well understood, their local and systematic nature suggest that squamous cell carcinoma-produced cytokines with proinflammatory and immunoregulatory activity may contribute to the pathogenesis of LSCC. In order to gain a better insight into the roles and relationships of the cytokines, we investigated serum IL-6, IL-10 and IL-12 concentrations in LSCC patients under baseline conditions and after surgery. In comparison with controls, all the patients had higher plasma IL-10 concentrations before surgical treatment (T0), while plasma IL-6 and IL-12 concentrations were higher in 22 (84.6%) and 24 patients (92.3%). The differences in plasma IL-6, IL-10 and IL-12 concentrations at T0 and T1 were statistically significant (p<0.001, p<0.0046 and p<0.011). Our finding suggest that plasma cytokines are overexpressed in LSCC patients. There was an independent increase in plasma IL-6 levels before and after surgical treatment. Furthermore, the up- and down-regulation of plasma IL-10 and IL-12 suggest a regulatory relationship between them.

The effect of intranasal cocaine and lidocaine on nitroglycerin-induced attacks in cluster headache.

The administration of nitroderivatives in cluster headache (CH) sufferers is the most reproducible experimental paradigm to induce spontaneous-like pain attacks. Previous uncontrolled studies have reported that the local use of anaesthetic agents in the area of the sphenopalatine fossa is able to extinguish nitroglycerin (NTG)-induced pain in CH. The present study, carried out according to a double-blind placebo-controlled design, included 15 CH patients, six with episodic CH (mean +/- SD age of 36.8+/-5.6 years), and nine with chronic CH (37.8+/-10.4 years). Patients had undergone a standard NTG test (0.9 mg sublingually), during which the intensity of pain was scored using a visuoanalogic scale (VAS, range 0-10). Nine patients (two with the episodic form, seven with the chronic form) experienced a typical, spontaneous-like attack on the usual side, occurring in all cases within 45 min. In these patients, the test was repeated with an interval of 2 days, and once pain intensity reached 5 on the VAS, a 10% solution of cocaine hydrochloride (1 ml, mean amount per application 40-50 mg), or 10% lidocaine (1 ml), or saline was applied using a cotton swab in the area corresponding to the sphenopalatine fossa, under anterior rhinoscopy. This was done in both the symptomatic and the non-symptomatic side, for 5 min. Treatments were always performed randomly, in separate sessions. All patients responded promptly to both anaesthetic agents, with complete cessation of induced pain occurring after 31.3+/-13.1 min for cocaine and 37.0+/-7.8 min for lidocaine (M+/-SD). In the case of saline application, pain severity increased thereafter, and extinction of the provoked attacks occurred with a latency of 59.3+/-12.3 min (P<0.01 and P<0.01 vs. cocaine and lidocaine, respectively, Mann-Whitney U-test). While further suggesting that the sphenopalatine ganglion participates in the mechanisms of pain, these findings indicate that the local administration of the anaesthetic agents cocaine and lidocaine is effective on NTG-induced CH attacks, and may be used in the symptomatic treatment of this disorder.

The organization of a counselling service for the families of patients undergoing bone marrow transplant.

The organisation of our Counselling Service for the relatives of patients undergoing bone marrow transplant is structured in various phases. During the first phase, contact is established with the relatives. The therapist introduces himself to the relatives when the patient is hospitalized and enters the sterile ward. The second phase consists in an illustration of the objectives and purpose of the Service. During this phase, two weekly interviews lasting approximately one hour are proposed, plus assessment tests (CFI, Camberwell Family Interview--and MMPI--Minnesota Multiphasic Personality Inventory). A third phase, which is often contextual to the presentation of the Service, consists in the recognition of the needs and suffering of the given relative. Here, the therapist acts as a receptacle for the anxieties and fear of death brought to him by the subject, thus empathetically accepting these states of mind and legitimizing them. Moreover, our model of intervention envisages the formation of support groups for the relatives of patients suffering from blood cancers. A further phase concerns the exchange of information between the two therapists helping, respectively, the relative and the patient, and also with medical and nursing staff. To further the same aims, for a number of years now, our Service has been organising intervention groups for nursing staff working at CTMO. The final phase is counselling, which takes place twice a week. There are two fundamental areas of intervention that must be taken into account for families of subjects suffering from blood cancers: providing information and offering support. From our experience, it emerges that the relatives need to obtain information on organic aspects of the illness and the transplant.

Immunohistochemical and molecular analysis of bax, bcl-2 and p53 genes in laryngeal squamous cell carcinomas.

BACKGROUND: We investigated the role of apoptosis and its potential alterations in laryngeal squamous cell carcinomas (LSCCs) by evaluating bax, bcl-2 and p53 protein expression in 50 cases and by characterising the molecular status of the bax and p53 genes. MATERIAL AND METHODS: p53 and bax gene mutations were investigated by means of PCR/SSCP and direct DNA sequencing, and bax, bcl-2 and p53 protein expression by means of immunohistochemistry. RESULTS: We identified p53 gene mutations in 17/50 cases (34%); p53 expression in 26 of the 50 cases (52%); bcl-2 expression in 5/50 cases (10%); bax expression in 32/47 cases (68%). 18/33 cases with a wild type p53 gene overexpressed p53 protein: 12 cases (approximately 66%) were bax+/bcl-2-. Of the remaining cases without p53 protein expression, seven cases (approximately 47%) were bax+/bcl-2-. CONCLUSIONS: Our observations suggest that the overexpression of p53 may contribute to the repression of bcl-2 and the induction of bax expression in LSCCs. However, the fact that a number of cases not expressing p53 did not present any clear up-regulation of bax or down-regulation of bcl-2 suggests that bcl-2 and bax may be regulated by various mechanisms other than p53.

[Occupational olfactory changes: diagnostic trends]. / Le alterazioni olfattive da causa professionale: orientamenti per la diagnosi.

Olfactory testing has been of minor interest in Occupational Health due to the lack of testing methods able to detect malingering. On the other hand there is evidence that occupational exposure to several, mainly neurotoxic, substances may result in olfactory damage. We have combined three different testing methods in one package in order to assure a forensic-degree level of results. The package consists of: 1. primary neuron functionality testing with a single olfactory stimulant; 2. olfactory-trigeminal discrimination testing with regular sniff-test; 3. odor identification score by Doty's UPSIT test. Final judgement of a link between olfactory system impairment and occupational exposure to chemicals requires a good knowledge of the present and past occupational exposure and of the general conditions of the patient. It requires collaboration between the Occupational Health specialist and the expert in Olfactology and may be completed with endoscopy, radiography and other specific controls. We suggest that a more extensive use of appropriate olfactory testing should be established at least for special risk groups of workers. This will not only detect occupational health damage that would otherwise have remained unknown, but can also furnish new information on the neurotoxic effects of many inhalable chemicals.

p53 and cyclinX D1 protein expression in carcinomas of the parotid gland.

BACKGROUND: p53 and cyclin D1 genes play a central role in the regulation of the G1 phase of the cell-cycle, and are frequently involved in head and neck tumorigenesis. MATERIALS AND METHODS: By means of immunohistochemistry, we retrospectively investigated the overexpression of cyclin D1 and p53 genes in a series of 28 parotid gland carcinomas. The immunohistochemical analysis was performed using the ABC method and the antibodies DCS6 (for cyclin D1) and CM1 (for p53). RESULTS: p53 was overexpressed in 12 (42.9%) and cyclin D1 in 6 cases (21.4%). No significant association was found between p53 or cyclin D1 expression and the evaluated clinicopathological parameters of tumor extension, clinical stage, and lymph node or distant metastases. Nevertheless, it is worth noting that all of the patients with a high expression of p53 died of their disease. CONCLUSIONS: The present data confirm the role of p53 abnormalities in the pathogenesis of salivary gland carcinoma and report, for the first time, the involvement of cyclin D1 gene in these tumors.

Cardiac malignant lymphoma: a case report.

A 76 year old man, who had undergone tonsillectomy for non Hodgkin's malignant lymphoma, died suddenly in rapidly evolving cardiogenic shock with electrocardiographic signs of acute lateral myocardial infarction. Post mortem examination showed three cardiac lesions, two in the left ventricle and one in the right atrium, corresponding to the "crista terminalis". Histologic examination of autopsy samples confirmed the presence of lymphoma in the heart, partially affecting the sino atrial node (NSA) but excluding other sites. There was no evidence of acute myocardial infarction in spite of the clinical signs and symptoms. An infarction-type electrocardiographic pattern associated with conduction disturbances in patients with lymphoma should suggest the possibility of cardiac localization of the disease.

Trigeminal evoked potentials in man: a new olfactory stimulation device.

The recording of olfactory evoked potentials in healthy humans, using a continuous flow olfactory stimulator, is described. A stimulator pushed inert gas (N2) in a continuous flow through the nose at a rate of 4 l/min. At fixed 30-second intervals, (32 times) the flow was replaced by an equal amount of CO2, a trigeminal stimulant. Each pulse lasted 200 ms. An electronic timing circuit triggered both the stimulator and the recorder. Signal acquisition was performed using an Evoked Potential Recorder (Nicolet Compact Four by Nicolet Biomedical Instruments), triggered by the stimulator. Using this stimulator device reliable olfactory evoked potentials can be recorded in a clinical setting. Since this is a non invasive technique which can be used to test olfactory function whether or not the patient cooperates, it is expected to become widely used, particularly in non collaborating patients and in those suspected of malingering.

Esthesioneuroblastoma: a general review of the cases published since the discovery of the tumour in 1924.

Esthesioneuroblastoma (ENB) arises from the neuroepithelium in the olfactory rim of the nasal cavity. It accounts for about 3% of all intranasal tumours. Reviews since the first description by Berger and Luc in 1924 never reported more than a hundred cases, stressing the rarity of the tumour. However, a thorough literature review revealed a total of 945 reported cases. In our search we found a total of 1,457 cases chronicled in the literature of which perhaps 487 were cited in more than one paper, bringing the total of reported cases to 945. Author cases accounted for 198 and therefore collaborative efforts accounted for 747 cases. Sex distribution was 53.36% male and 46.64% female. Kadish classification was applied to 553 cases revealing 103 (18.29%) class A cases, 182 (32.33%) class B and 278 (49.38%) class C cases. This distribution was generally stable through the decades. Treatment could be classified in 898 cases. It consisted of surgery alone in 25.17% (226 cases), radiotherapy alone in 18.37% (165 cases), combined surgery and radiotherapy in 43.21% (388 cases) and chemotherapy in 13.2% (119 cases), followed in 11 cases (1.22%) by bone marrow transplant. In the reported cases an overall follow up could be evaluated in 477 cases, while in only 234 cases a five-year follow up was done. The outcome was 68.38% alive and disease free, 12.82% alive with disease and 18.80% dead. From these 20.51% had surgery only, 11.11% radiotherapy and 68.38% combined surgery and radiotherapy. The best survival rates were obtained by combined therapy (72.5% vs. 62.5% surgery alone and 53.85% radiotherapy alone). Death rates were highest after radiotherapy alone (30.77% versus 18.75% in combined therapy and 12.50% after surgery alone). In conclusion, ENB is a rare but not exceptional tumour. It is best treated with combined surgery and radiotherapy. Unfortunately early diagnosis is still uncommon and no significant changes to the proportions of Kadish classes at first diagnosis have been noted in recent decades. A greater awareness of the tumour and earlier diagnosis seems the major focus for future research.

PCNA--a cell proliferation marker in vocal cord cancer. Part II: Recurrence in malignant laryngeal lesions.

Laryngeal squamous cell carcinoma constitutes the most frequent carcinoma found in the head and neck region. A precise prediction for recurrence potential cannot be done on site, treatment and histologic grading. Since Proliferating Cell Nuclear Antigen (PCNA) and DNA-cytometry have shown a good correlation between premalignant lesions and their progressive potential towards full-fledged carcinoma in the larynx as described in part I of this work, we have analyzed the PCNA index and DNA cytometry in specimen taken from vocal chord carcinomas with a 5-year follow-up, in order to assess its relationship with the presence or absence of tumour progression. 42 cases with (21) and without (2) recurrence have been examined. The DNA-index ranged from 1.01 to 1.43 (mean 1.10) in the group without and from 1.02 to 1.59 (mean 1.38) in the group with recurrent carcinoma (p = 0.002). The PCNA-index ranged from 0.00% to 18.90% (mean 6.97%) in the nonrecurrent group and from 0.00 to 3g.50% (mean 16.35%) in the patients with recurrence (p = 0.001). Both indices also correlated in a highly significant way. From these data emerges a highly significant correlation between the cytometric indices of cell proliferation and PCNA immunostaining. Furthermore the high correction between PCNA and DNA-index is of special interest for single case assessment. High DNA aberration and PCNA-index in vocal chord carcinoma may indicate a higher cellular aggressiveness of the tumour, resulting in a greater overall risk of metastases and local recurrences. Our results support the thesis that the indices of cellular proliferation within some cancers can define subsets of patients of high risk and help in isolating a population in which a more aggressive clinical protocol may be proposed.

PCNA--a cell proliferation marker in vocal chord cancer. Part I: Premalignant laryngeal lesions.

Laryngeal hyperkeratotic lesions can progress to fully developed malignant carcinoma in some cases. These premalignant lesions are proliferative disorders whose potential for further tumour progression is perhaps difficult to assess by mere histology. Immunostaining with PCNA, a protein correlated with cell proliferation, has been used to study tissue behavior in 30 cases of premalignant laryngeal vocal chord lesions treated by epithelial stripping in microlaryngoscopy, 15 of whom had no progression and 15 had recurrence and final development of full malignancy. The results showed a statistically significantly higher PCNA-index in the cases which underwent further tumour progression towards malignancy. PCNA testing may thus be suggested as a marker for tumour progression potential and help in determining clinical treatment choices.

Carcinoma of the vocal cord. Results after subperichondral cordectomy.

The clinical outcome of 110 patients operated upon by subperichondral cordectomy between 1982 and 1992 for T1s/T1a, NO, MO carcinomas of the vocal cord has been evaluated in this longitudinal epidemiological study. Patients have been followed up until the end of 1993 by examinations done once a month (first year), every three months (years 2 to 4), every six months (years 5 to 8), and then once a year. Life-tables have been computed according to Kaplan and Meier and raw survival has been 90.0%. Considering only the mortality due to any type of neoplastic disease, the survival reached 93.6%. Finally, considering only deaths due to recurrences or metastases of the primary tumor, the survival rate was 95.5%. Mortality after the first recurrence was 27.3%, after a second recurrence 50.0%. A correlation between number of cigarettes smoked and the risk of recurrence of the tumor could be observed (p < 0.01), while gross appearance and histologic grading of the vocal cord carcinoma proved to be uncorrelated with the risk of recurrences.

[Epidemiologic longitudinal study of a population of workers of the state railroads exposed to noise]. / Studio epidemiologico longitudinale di una popolazione di lavoratori delle Ferrovie dello Stato esposta al rumore.

Loss of hearing due to chronic noise exposition is a well-known pathology and many nations have regulations for the protection of workers with periodical audiometry and ONT check-ups. For the last 15 years, the authors have followed state railway workers of the Milan area who were professionally exposed to noise. All were periodically examined by ONT specialists and submitted to audiometry. The findings underscore the importance of these periodical examinations and their efficacy for the prevention of hearing loss due to noise.

Electronmicroscopic demonstration of HPV in oral warts.

Human Papilloma Virus (HPV) has been demonstrated in a series of benign proliferative lesions of the skin and the mucosae. The virus has also been found in verrucous laryngeal carcinoma and carcinomas of the oral cavity and other organs. DNA hybridization techniques have classified, the HPV into 51 types, some of which seem to be associated with specific lesions. In order to study the intracellular distribution of HPV, we performed ultrastructural analysis with the electron microscope on 14 specimens taken from 7 patients by large excisional biopsy, which had been histologically classified as "fibropapilloma". From each patient specimens were taken from both the clinically evident lesion and the clinically normal surrounding mucosa. The specimens were fixed with glutaraldehyde, washed with cacodylate buffer, post-fixed with potassium ferrocyanide reduced-osmium tetroxide, block stained with uranyl acetate and embedded in EPON 812. The tissues underwent to amylase digestion before the electron microscopic examination. We found a large number of viral particles in both nuclei and cytoplasm, without forming crystal array structures as described typically for the virus of the verruca vulgaris (HPV-2). No significant differences were found between the cells derived from the clinical lesion and those derived from the surrounding mucosa. The passage of viral particles from infected to not yet infected cells through the intercellular space was observed. Of particular interest, we found a high intracytoplasmatic presence of the virus and its clear abundance in the cells surrounding the clinical lesion.

Electron microscopic detection of human papillomavirus particles in oral proliferative lesions.

Human papilloma virus (HPV) has been demonstrated in a series of benign proliferative lesions of skin and mucosae. To prove the distribution of HPV in the oral proliferative lesions at the ultrastructural level, we performed electron microscopic analysis of 10 specimens taken from 5 patients through large excisional biopsy. All of them were diagnosed pathologically as fibropapilloma. In each patient, specimens were taken from both clinically evident proliferative lesions and clinically normal surrounding mucosa. Obtained specimens were fixed in a glutaraldehyde solution and processed for routine ultrathin sectioning. Before electron microscopic observation, the tissue sections on copper grids were subjected to amylase digestion of glycogen granules. Spherical viral particles of 40-55 nm in diameter were detected the non-keratinized epithelial cells in all specimens examined. Of particular interest were the large amounts of viral particles found in the cytoplasmic matrix and nuclei (especially on their chromatin masses) of the cells in intermediate and surface layers, which did not form a crystal array. All the membranous cell organelles of epithelial cells were, however, devoid of viral particles. Some viral particles were distributed in the extracellular spaces of an intermediate layer. Viral particles were hardly observed in the cells of a basal/suprabasal and prickle cell layers. There were no significant differences in the HPV distribution between the cells derived from the proliferative lesion and those derived from the surrounding normal mucosa.

[Ultrastructural demonstration of the human papilloma virus (HPV) in oral proliferative lesions]. / Dimostrazione ultrastrutturale del virus del papilloma umano (HPV) in lesioni proliferative orali.

The Human Papilloma Virus (HPV) has been found in a series of benign proliferative lesions of the skin and mucosa. The virus has also been found in verrucous laryngeal carcinoma and carcinomas of the oral cavity and other organs. DNA hybridization techniques have made it possible to classify 51 types of HPVs, some appearing to be associated with specific lesions. In order to study the intracellular distribution of HPVs, an ultrastructural morphological analysis was performed with an electron microscope on 10 specimens taken from 5 patients. The specimens were obtained through large excisional biopsy, histologically classified as "fibropapilloma". For each patient specimens were taken from the clinically evident lesion and from the surrounding clinically normal mucosa. The specimens were fixed in glyceraldehyde, washed in a cacodylate buffer, post-fixed in potassium ferrocyanide reduced-osmium tetroxide, stained with uranyl acetate and included in EPON 812. The tissue was subjected to amylase digestion prior to electron microscope examination. A great number of viral particles were found in both the nucleus and the cytoplasm, without forming crystal array structures as typically described for the verruca vulgaris virus (HPV-2). No significant differences could be found between the cells derived from the clinical lesion and those derived from the surrounding mucosa. The passage of viral particles from infected to as yet uninfected cells through the intercellular space could be seen. The high intracytoplasmatic presence of the virus, and its clear abundance in those cells surrounding the clinical lesion, is felt to be of special interest.

[Alphafetoprotein in serum of pregnant women. A new assay and practical aspects in prenatal screening for fetal neural tube malformations (author's transl)]. / Alphafetoprotein im Schwangerenserum--Eine neue Messmethode und praktische Aspekte bei der prnatalen Suche nach kindlichen Neuralrohrmissbildungen.

A new radioimmunoassay for alphafetoprotein was tested with 440 sera for it's suitability in the screening of pregnant women for fetal neural tube malformations. The test was reliable and it's execution simple and little time consuming. The test allowed to establish the normal range during pregnancy as well as to detect typical deviations from normal in cases of abortion and anencephaly. Results of mothers with diabetes or elevated transaminase activities were in the normal range. The practicability and acceptable cost of such a test render now possible, and at the same time urgent, a regional screening of women from the 15 th to the 18th week of pregnancy. Only in this way, the detection rate of fetal neural tube malformations can be measured and the usefulness of a general screening of all pregnant women can be evaluated.