[Assessment of the presence of mutations of the epidermal growth factor receptors in tumors of various histologies]. / Valutazione della presenza di mutazioni del recettore dell'epidermal growth factor in tumori di varia istologia.

Recent reports from US and Japan have established that mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (Egfr) occur in a subset of patients with lung cancer that respond to therapy with gefitinib, a TK inhibitor. To gain further insights into the role of Egfr in carcinogenesis of lung and tumors of diverse histology, that are currently under investigation with drugs of the same class, we have taken in examination a panel of tumors consisting in 110 pulmonary adenocarcinomas, 40 pulmonary squamous carcinomas, 40 gastric adenocarcinomas and 40 colorectal adenocarcinomas. The sequence analysis of exon 19 and 21 of the Egfr has allowed the identification of 10 cases exhibiting specific deletions in exon 19 and 1 case with point mutation in a conserved residue in exon 21. All Egfr mutations occur specifically in lung adenocarcinomas while tumors of different histology result unaffected. The rate of mutation affecting these other tumors is either very rare, involves different domains of the receptor or other tyrosine kinases. The molecular analysis of the Egfr gene can help identify patients that will benefit from gefitinib therapy.

[Association of mutations of K-RAS oncogene and deletions of 18Q with lymph node metastasis of colorectal cancer]. / Associazione di mutazioni dell'oncogene K-RAS e delezioni del 18Q con metastasi linfonodali nel cancro del colonretto.

Molecular characterization of gastrointestinal cancer has greatly helped the definition of the key steps of the malignant transformation process and made it the best understood among the malignant cancers. Genetic influences on prognosis may have important implications for the management of the disease and help to design patient-tailored therapy. In order to acquire additional knowledge on this issue we have commenced an institutional study with the aim to identify the most frequent molecular alterations and make a correlation with the conventional histopathological parameters.

¿Podemos predecir clínicamente la cervicoartrosis? / Can we predict cervicoarthrosis clinically?

Fundamento: desconocimiento del valor de la exploración física cervical. Objetivo: establecer la utilidad y predictividad de algunas maniobras cervicales en relación a la presencia de cervicoartrosis. Diseño: estudio descriptivo transversal con muestreo sistemático. Pacientes y métodos: sobre una muestra de 100 personas que se realizaban una exploración radiológica cervical en el Servicio de Radiodiagnóstico de la DAP Cornellà Centre (enero a mayo 2000, muestreo sistemático), evaluamos para cada una de las personas dos pruebas de exploración física (test de rotación cervical medida con goniómetro estático y test barbilla-pecho), comparando esta evaluación con la obtenida en la exploración radiológica, efectuada por un radiólogo que desconocía el resultado de la exploración clínica. Resultados: cuando la rotación cervical lateral era menor de 56º, obteníamos un valor predictivo positivo del 85 por ciento, una sensibilidad del 81 por ciento y una especificidad del 61 por ciento. La valoración clínica cervical con estos dos tests se correlacionaba de forma significativa con la gravedad radiológica, sin llegar a hacerlo con la concordancia (índice Kappa). Una maniobra de rotación cervical normal (igual o mayor a 70º) excluía la presencia de cervicoartrosis. Conclusiones: el test de rotación cervical tiene relevancia clínica y la combinación con otras maniobras exploratorias permite un cribado clínico previo y complementario a la evaluación radiológica (AU)

The tetraheme cytochrome c NrfH is required to anchor the cytochrome c nitrite reductase (NrfA) in the membrane of Wolinella succinogenes.

The electron-transport chain that catalyzes nitrite respiration with formate in Wolinella succinogenes consists of formate dehydrogenase, menaquinone and the nitrite reductase complex. The latter catalyzes nitrite reduction by menaquinol and is made up of NrfA and NrfH, two c-type cytochromes. NrfA is the catalytic subunit; its crystal structure is known. NrfH belongs to the NapC/NirT family of membrane-bound c-type cytochromes and mediates electron transport between menaquinol and NrfA. It is demonstrated here by MALDI MS that four heme groups are attached to NrfH. A Delta nrfH deletion mutant of W. succinogenes was constructed by replacing the nrfH gene with a kanamycin-resistance gene cartridge. This mutant did not form the NrfA protein, probably because of a polar effect of the mutation on nrfA expression. The nrfHAIJ gene cluster was restored by integration of an nrfH-containing plasmid into the genome of the Delta nrfH mutant. The resulting strain had wild-type properties with respect to growth by nitrite respiration and nitrite reductase activity. A mutant (stopH) that contained the nrfHAIJ locus with nrfH modified by two artificial stop codons near its 5' end produced wild-type amounts of NrfA in the absence of the NrfH protein. NrfA was located exclusively in the soluble cell fraction of the stopH mutant, indicating that NrfH acts as the membrane anchor of the NrfHA complex in wild-type bacteria. The stopH mutant did not grow by nitrite respiration and did not catalyze nitrite reduction by formate, indicating that the electron transport is strictly dependent on NrfH. The NrfH protein seems to be an unusual member of the NapC/NirT family as it forms a stable complex with its redox partner protein NrfA.

Validation of a questionnaire to evaluate the quality of life of nonprofessional caregivers of dependent persons.

AIM OF THE STUDY: To determine the acceptance, validity and reliability of the questionnaire for assessing the type of informal care that caregivers of dependent people give and the effects this care might have on the health of the carer. BACKGROUND: In Spain, the formal health care system provides 12% of the total time dedicated to health care, the remaining 88% is carried out by the informal system within the home environment. The act of caring has effects on various areas of the life of the carer and on family functioning. This makes clear the existing risk when the principal carer becomes a secondary nurse. METHODS: This research was a cross-sectional design, carried out in municipalities in the province of Barcelona (Spain) from January to December 1997. The subjects of the study were 240 caregivers of dependent people. The questionnaire (ICUB97Copyright ) is based on the Virginia Henderson's 14 Needs nursing model. The validity of the content was assessed through the consensus of a group of experts, validity of design by means of comparison with the hypotheses. RESULTS: Test-retest reliability was completed for the three parts of the questionnaire and the kappa index values was 0.89, 0.80 and 0.75 for each part. The higher the level of dependency of the person cared for, the more care tasks the carer had to perform. A correlation coefficient of 0.58 was obtained for the Barthel Index (P < 0.001) and 0.53 on the Philadelphia Index (P < 0.001). The care tasks performed by the carer showing greatest correlation with dependency level were; help with elimination (r=0.73, P < 0.001), help with feeding (r=0.55, P < 0.001) and help in personal development (r=0.55, P=0.001). CONCLUSIONS: This questionnaire provides a reliable and valid instrument for measuring the care given by caregivers to dependent people, to meet their basic needs as well as for assessing the needs of the carers who experience problems by the act of caring. It is therefore important for nursing practice, to have a validated instrument available for identifying the tasks performed by family carers and the effects on their health.

[Impact of caregiving on the health of family caregivers]. / Impacto del hecho de cuidar en la salud de los cuidadores familiares.

OBJECTIVES: To identify the type of care provided by informal carers of dependent persons and the repercussions this care might have on the health of the carers, and to find the characteristics of both informal carers and cared-for people. DESIGN: An observational cross-sectional study. SETTING: This study was conducted in various towns in the province of Barcelona between January and December 1997 in primary health care. PARTICIPANTS: Those taking part were 240 informal carers (IC) for dependent persons. MEASUREMENTS AND MAIN RESULTS: The ICUB 97 questionnaire was the data-gathering instrument. It was validated previously by the research team and based on the fourteen needs of the Virginia Henderson nursing model. The questionnaire was filled in at a personal interview. The level of dependence of people cared for was evaluated with the Barthel and Philadelphia Geriatric Center indices. The analysis of the results reflected that the greater the level of dependence of the person cared for, the more care is provided by the carer. The main repercussions of caring on the health of the carers were: back pain (73%), tiredness (72%), reduced leisure time (73%), insomnia (65%), anxiety (72%) and changes in family life (54%). Repercussions that correlated most closely with the fact of caring were: sleep disorders, family economy, personal development and leisure, middle age and having few educational qualifications. CONCLUSIONS: Most carers are middle-aged women performing multiple care tasks. This work causes their quality of life to deteriorate.

Impacto del hecho de cuidar en la salud de los cuidadores familiares / Impact of caring on the health of family carers

Objetivos. Identificar el tipo de cuidados que prestan los cuidadores informales de personas dependientes y las repercusiones que pueden ocasionar estos cuidados en la salud del cuidador, así como conocer las características de los cuidadores informales y las de las personas cuidadas. Diseño. Se presenta un estudio observacional de tipo transversal. Emplazamiento. Este estudio se ha realizado en varios municipios de la provincia de Barcelona, de enero a diciembre de 1997, en el ámbito de la atención primaria de salud. Participantes. Los sujetos de estudio fueron 240 cuidadores informales (CI) de personas dependientes. Mediciones y resultados principales. El instrumento de recogida de datos fue el cuestionario ICUB 97, validado previamente por el equipo investigador, que está basado en las 14 necesidades del modelo de enfermería de Virginia Henderson. El cuestionario se cumplimentó a través de entrevista personal. El nivel de dependencia de las personas cuidadas se evaluó con los índices de Barthel y Philadelphia Geriatric Center. El análisis de los resultados reflejó que cuanto mayor es el nivel de dependencia de la persona cuidada, más cuidados presta el cuidador. Las principales repercusiones que ocasiona el hecho de cuidar en la salud de los cuidadores son: dolor de espalda (73 por ciento), cansancio (72 por ciento), disminución del tiempo de ocio (73 por ciento), insomnio (65 por ciento), ansiedad (72 por ciento) y alteraciones en la vida familiar (54 por ciento), y las que más se correlacionan con este hecho de cuidar son: alteraciones del sueño, economía, desarrollo personal y ocio. Conclusiones. La mayoría de personas cuidadoras son mujeres de mediana edad que realizan múltiples tareas de cuidados y dicha labor repercute en un deterioro de su calidad de vida (AU)

Toxic adenoma and papillary thyroid carcinoma in a patient with Graves' disease.

A case of a very rare association of toxic adenoma and papillary carcinoma with Graves' disease is presented. A 34-year-old woman developed Graves' disease with mild ophthalmopathy. An ultrasound revealed diffuse thyroid enlargement with a hypoechoic pattern and a hypoechoic nodule with regular edges of 1.6 cm in diameter at the lower pole of the left lobe. A thyroid 131I scintiscan showed a diffuse and homogeneous 131I distribution. The 131I uptake (RAIU) was elevated. One year later, while still on a low dose of methimazole, the patient had a recurrence of hyperthyroidism following an iodine load from a contrast agent. A further thyroid ultrasound confirmed the previously described pattern but showed a new hypoechoic nodule of 1.1 cm with irregular edges in the left lobe. A thyroid 131I scintiscan this time demonstrated a hyperactive area localised in the larger nodule and a lower diffuse uptake of the remaining tissue. Because of the worsening of the symptoms of hyperthyroidism, the patient had a left lobectomy. On histological examination, the larger nodule was well encapsulated and showed the characteristics of a hyperfunctioning follicular adenoma. The smaller nodule was a typically unencapsulated papillary carcinoma. Several other microfoci of papillary carcinoma were also found in the adjacent tissue. Completion of thyroidectomy was therefore performed, followed by 131I ablative therapy and thyroxine suppressive treatment. This observation suggests that the chronic abnormal stimulation of the thyroid gland by the thyroid-stimulating antibody (TSAb) may facilitate the neoplastic transformation of the thyrocytes in individuals with a critical genetic background.

The single cysteine residue of the Sud protein is required for its function as a polysulfide-sulfur transferase in Wolinella succinogenes.

The periplasmic Sud protein which is induced in Wolinella succinogenes growing by polysulfide respiration, has been previously proposed to serve as a polysulfide binding protein and to transfer polysulfide-sulfur to the active site of polysulfide reductase [Klimmek, O, Kreis, V., Klein, C., Simon, J., Wittershagen, A. & Kröger, A. (1998) Eur. J. Biochem. 253, 263-269.]. The results presented in this communication suggest that polysulfide-sulfur is covalently bound to the single cysteine residue (Cys109) of the Sud monomer, and that Cys109 is required for tight binding of polysulfide-sulfur and for sulfur transfer. A modified Sud protein [(C109S)Sud-His6] in which the cysteine residue was replaced by serine, did not catalyze sulfur transfer from polysulfide to cyanide and did not stimulate electron transport to polysulfide, in contrast to Sud-His6. The polysulfide-sulfur bound to (C109S)Sud-His6 was fully removed upon dialysis against sulfide. After this treatment, Sud-His6 retained one sulfur atom per monomer; thiocyanate was formed upon addition of cyanide to the preparation. After incubation of Sud-His6 with polysulfide, a proportion of the Sud-His6 monomers carried one or two sulfur atoms, as shown by matrix-assisted laser desorption ionization mass spectrometry. The sulfur atoms were absent from monomers derived from Sud-His6 treated with cyanide and from (C109S)Sud-His6 incubated with polysulfide.

Recurrent postinfantile syncytial giant cell hepatitis after orthotopic liver transplantation.

Syncytial giant cell hepatitis is a severe form of hepatitis characterized by diffuse giant cell transformation of hepatocytes. The disease may evolve to chronic cholestatic cirrhosis necessitating liver transplantation. We report the case of an adult liver transplant recipient presenting with early recurrent disease without concomitant clinicobiochemical syndrome. Early recurrence of giant cell hepatitis after liver transplantation favors the hypothesis of a transmissible agent as the etiology of the disease. Routine follow-up liver biopsy is necessary in these cases in order to gain more information about the precise incidence and aggressivity of disease recurrence in the allograft.

High-dose cyclophosphamide, etoposide and BCNU (CVB) with autologous stem cell rescue in malignant lymphomas.

Eighteen patients with malignant lymphoma, 10 non-Hodgkin's and 8 Hodgkin's, were treated with high-dose CVB (cyclophosphamide 4 x 1.5 g/m2, etoposide 4 x 250-400 mg/m2, carmustine 4 x 150-200 mg/m2), followed by autologous peripheral blood stem cells (PBSC, 13 patients) or bone marrow (BM, 5 patients) transplantation. At the time of autograft 6 patients were in complete remission (CR), 3 in partial remission (PR) and 5 in relapse (4 sensitive, 1 resistant), whereas 4 had progressive disease. All CR patients had poor prognostic features at presentation. PBSC were collected at the time of rapid hematologic recovery after intense chemotherapy by means of a cell separator. All patients engrafted. Median time to achieve > or = 0.5 x 10(9)/l polymorphonuclear cells (PMN) and > or = 50 x 10(9)/l platelets was 13 days for both cell types in PBSC autografted patients, versus 20 and 28 days respectively in BM autografted patients. A significant advantage of PBSC over BM was found in terms of time needed to recover either PMN > or = 0.5 and PMN > or = 1 x 10(9)/l (p = 0.01). Autograft-related toxicity consisted mainly of moderate severity interstitial pneumopathy (3 patients), and veno-occlusive disease (1 patient) that resolved completely. Of the 12 patients autografted with detectable disease, 6 (50%) obtained a CR. Seven out of 18 autografted patients (39%) had disease progression within 1 to 5 months of autograft. The projected progression-free survival is over 50% at 4 years and it was significantly longer in patients with sensitive disease than in those with resistant disease (p = 0.01). The efficacy and the low toxicity of CVB suggest that autograft with PBSC may be proposed for the primary treatment of poor prognosis malignant lymphomas.

Alpha interferon treatment of chronic hepatitis C in beta-thalassaemia.

In this open, pilot study, interferon (IFN) alpha-2b seemed effective in the treatment of hepatitis C virus (HCV) infection in patients with beta-thalassaemia. In seven of nine patients who completed the study alanine aminotransferase activities returned to normal, and a completely stable response 24 months after treatment was seen in five. Liver biopsy specimen showed a clear reduction in portal, periportal, and lobular necroinflammation in all five cases. Three patients stopped treatment early because of side effects.

alpha-Interferon treatment of chronic hepatitis C in young patients with homozygous beta-thalassemia.

BACKGROUND: Chronic infection with the hepatitis C virus (HCV) and iron overload are the main causes of chronic liver disease in subjects with homozygous beta-thalassemia (HBT). Iron overload can be counteracted by intensive chelation. alpha-interferon reduces viremia and necroinflammation in patients with chronic HCV hepatitis. METHODS: To assess the effectiveness and safety of alpha 2b-Interferon (IFN), we enrolled in an open pilot trial of treatment 12 patients with HBT and biopsy-proven anti-HCV positive chronic hepatitis. IFN was given at a dose of 5 MU/m2 thrice weekly for 8 weeks, then 3 MU/m2 thrice weekly for 18 weeks. Patients were followed up to 24 months after stopping treatment when a second liver biopsy was performed in subjects with sustained response (normal ALT during follow-up). RESULTS: Two patients discontinued IFN at 7 weeks because of haemolytic anemia and one after 8 weeks due to persistent fever. Among 9 subjects completing the protocol, 5 normalized ALT while on treatment and a further 2 within two months after stopping IFN. A sustained response was obtained altogether in 5 patients, since ALT relapsed in 2 responders. None of the 3 subjects who discontinued IFN and of the 2 patients who did not respond to treatment normalized ALT over a 24 months follow-up. Post-treatment liver histology in long-term responders showed a reduction of portal, periportal and lobular necroinflammation, while siderosis was essentially unchanged. CONCLUSIONS: Although the pattern of response to IFN in HCV-infected subjects with HBT might differ from that of non-thalassemics, due to peculiar side effects and delayed response, the drug appears to be effective and deserves further investigation.

Expression of leukocyte adhesion molecules in the liver of patients with chronic hepatitis B virus infection.

Virus-specific T-cell responses are believed to be involved in the pathogenesis of liver cell injury secondary to hepatitis B virus infection. In this study, liver biopsy specimens from patients with chronic hepatitis B virus infection were analyzed for expression of two major pathways of adhesion used by cytotoxic T cells to interact with target cells. The lymphocyte function-associated antigen 3 was found preferentially expressed on hepatocytes of patients with active hepatitis B virus replication, whereas the expression of the intercellular adhesion molecule 1 on hepatocytes seemed more closely related with inflammatory activity. Adhesion molecules were also highly expressed on T lymphocytes found in areas of piecemeal and spotty necrosis, indicating the presence of antigen-specific "memory" T cells at the site of hepatocellular injury. This study suggests that the expression of the lymphocyte function-associated antigen 3 on hepatocytes may be important for viral elimination. The coordinate expression of the intercellular adhesion molecule 1 may regulate inflammatory response and enhance viral antigen presentation to T cells. Conversely, the absence of hepatocyte adhesion molecules might be a favorable factor for viral persistence.

Natural history of congestive gastropathy in cirrhosis. The Liver Study Group of V. Cervello Hospital.

In a prospective study of the natural history of congestive gastropathy, 212 consecutive cirrhotic patients (75 treated with sclerotherapy) were included. Mean follow-up was 46 months. Mild gastropathy (mosaiclike pattern) was found in 110 patients and severe gastropathy (granular mucosa with cherry spots) was found in 20. Prevalence of Helicobacter pylori, formerly Campylobacter pylori, was 50% in patients without, 43% in those with mild, and 28% in those with severe gastropathy. Congestive gastropathy was significantly more frequent in patients treated with sclerotherapy (83% vs. 50%, P less than 10(-5)). Sixty-month actuarial proportions of patients free of anemia (in the absence of hematemesis or melena), were 17% with severe, 62% with mild, and 93% without gastropathy (P less than 10(-8]. Corresponding figures for overt bleeding were 25%, 73%, and 87% (P less than 10(-7], whereas those for survival were 46%, 72%, and 85% (P = 0.0005), respectively. A multivariate regression analysis supported the following conclusions: (a) sclerotherapy and the presence of large esophageal varices significantly increase the risk of congestive gastropathy, which (b) is a significant risk indicator of both chronic and overt bleeding but does not independently affect survival.

Procollagen type I production by hepatocytes: a marker of progressive liver disease?

The presence of collagen-producing cells and its relation to disease activity were determined in cryostat liver tissue sections from subjects with active cirrhosis (n = 15), inactive cirrhosis (n = 5), chronic persistent hepatitis (n = 8), or normal histology (n = 3) by means of an immunofluorescence technique using a monoclonal antibody to the carboxy-terminal domain of procollagen type I (anti-Pc). In all patients with active cirrhosis hepatocytes showed a strong intracellular staining with anti-Pc; in 4 of them bileducts also showed a membrane-like reaction. By contrast, tissue sections from chronic inactive liver disease and normal liver were essentially negative. These findings suggest that in chronic liver disease hepatocytes and sometimes biliary epithelium produce collagen and that production is related to disease activity. The detection of active production of procollagen type I by hepatocytes could become a useful marker of progressive liver disease.

Phenoxy herbicides and soft-tissue sarcomas in female rice weeders. A population-based case-referent study.

A population-based case-referent study was conducted in an area of northern Italy where rice growing is the predominant agricultural activity and phenoxy herbicides have been used since 1950. Manual rice weeding was formerly performed by a seasonal female working population; in the early 1950s these women were concurrently exposed to chemical herbicides. Sixty-eight persons representing incident and histologically revised cases (31 women) and 158 population referents (73 women) were interviewed. The cases were histologically confirmed independently by two blinded pathologists, and exposure to phenoxy herbicides was assessed by two blinded pesticide researchers. An age-adjusted odds ratio of 0.91 was found for the living men (with suspect exposures; no man diagnosed as a case had been exposed with certainty to phenoxy herbicides). Among the living women the relative risk was 2.7 (90% confidence interval 0.59-12.37), and it further increased when attention was restricted to women exposed in the whole 1950-1955 period and to younger age groups.