A novel versatile precursor suitable for 18 F-radiolabeling via "click chemistry".

As an effort to improve 18 F-radiolabeling of biomolecules in method robustness and versatility, we report the synthesis and radiolabeling of a new azido precursor potentially useful for the so-called "click reaction," in particular the ligand-free version of the copper(I)-catalyzed alkyne-azide cycloaddition. The new azido precursor may help to overcome problems sometimes exhibited by most of the currently used analogues, as it is safe to handle and it displays long-term chemical stability, thus facilitating the development of new radiolabeling procedures. Moreover, the formed 18 F-labeled 1,2,3-triazole is potentially metabolically stable and could enhance the in vivo circulation time. The above azido precursor was successfully radiolabeled with 18 F, with 51% radiochemical yield (nondecay-corrected). As a proof of concept, the 18 F-labeled azide was then tested with a suitable alkyne functionalized aminoacid (l-propargylglycine), showing 94% of conversion, and a final radiochemical yield of 27% (>99% radiochemical purity), nondecay-corrected, with a total preparation time of 104 minutes.

Quinolinic acid induced neurodegeneration in the striatum: a combined in vivo and in vitro analysis of receptor changes and microglia activation.

PURPOSE: Huntington's disease (HD) is a progressive neurodegenerative disorder, which is characterised by prominent neuronal cell loss in the basal ganglia with motor and cognitive disturbances. One of the most well-studied pharmacological models of HD is produced by local injection in the rat brain striatum of the excitotoxin quinolinic acid (QA), which produces many of the distinctive features of this human neurodegenerative disorder. Here, we report a detailed analysis, obtained both in vivo and in vitro of this pharmacological model of HD. MATERIALS AND METHODS: By combining emission tomography (PET) with autoradiographic and immunocytochemical confocal laser techniques, we quantified in the QA-injected striatum the temporal behavior (from 1 to 60 days from the excitotoxic insult) of neuronal cell density and receptor availability (adenosine A(2A) and dopamine D(2) receptors) together with the degree of microglia activation. RESULTS: Both approaches showed a loss of adenosine A(2A) and dopamine D(2) receptors paralleled by an increase of microglial activation. CONCLUSION: This combined longitudinal analysis of the disease progression, which suggested an impairment of neurotransmission, neuronal integrity and a reversible activation of brain inflammatory processes, might represent a more quantitative approach to compare the differential effects of treatments in slowing down or reversing HD in rodent models with potential applications to human patients.

Improved synthesis and radiolabeling of [11C]MP4A, a suitable ligand for the investigation of the cholinergic system using PET.

An improved synthesis of the precursor acetic acid-piperidine-4-yl ester by acetylation of 4-hydroxypiperidine hydrochloride in anhydrous chloroform was developed. A procedure for fast evaluation and characterization of products originated by acetylation of the 4-piperidinol using LC-APCI/MS with an acetonitrile-water gradient method on a Merck Purosphere RP-18 column was also developed. The highly purified precursor allowed the production of [11C]MP4A for PET studies of acetylcholine neurotransmission system. The tracer was produced with >98% radiochemical purity, with yields ranging 20-60% (decay-corrected) from EOB.

Evaluation of three quinoline-carboxamide derivatives as potential radioligands for the in vivo pet imaging of neurodegeneration.

The peripheral-type benzodiazepine receptors (PBRs) are only minimally expressed in normal brain parenchyma, where they are primarily localized in glial cells. Their basal expression rises in different neurodegenerative disorders, due to the presence of infiltrating inflammatory cells and activated microglia. [11C]PK11195, a selective PBR antagonist, has been used for the in vivo PET monitoring of neurodegeneration in clinical observations. We recently developed and labeled with carbon-11 three new carboxamide derivatives: [11C]VC193M, [11C]VC195 and [11C]VC198M. Aim of this study was to evaluate these ligands for the in vivo measuring of PBRs expression in neurodegenerations and compare their kinetic behavior with that of the reference tracer [11C]PK11195. Radioligands were evaluated in a preclinical model of Huntington's disease consisting in the monolateral striatal injection of quinolinic acid (QA). Activated microglia and astrocytic gliosis was present only within the affected striatum. A concomitant increase in radioactivity accumulation was observed for all the tracers examined (P<0.01). Among the new compounds, [11C]VC195 showed higher levels of lesioned/unlesioned striatum ratios (3.28+/-0.44), in comparison with [11C]VC193M and [11C]VC198M (2.69+/-0.53 and 1.52+/-0.36, respectively), but slightly inferior to that observed for [11C]PK11195 (3.76+/-1.41).In conclusion, the results of the study indicate that [11C]VC195 is a promising candidate for in vivo PET monitoring of neurodegenerative processes but its in vivo behavior overlap that of [11C]PK11195.

Monitoring of infections by protozoa of the genera Nematopsis, Perkinsus and Porospora in the smooth venus clam Callista chione from the North-Western Adriatic Sea (Italy).

Marketable smooth venus clams Callista chione from natural banks of Chioggia (Venice) and Goro (Ferrara), North-Western Adriatic Sea (Italy), were examined for protozoan parasites from November 1996 to November 1998. Out of the 375 bivalves examined, 149 (39.7%) were infected by Nematopsis sp. and 325 (86.7%) by Porospora sp. Oocysts of Nematopsis sp. were present with a prevalence that varied from 100% in November 1996 to 5% in June 1998; cystic and naked sporozoites of Porospora sp. were very common, with a prevalence of 100%. Out of the 229 bivalves examined between January and November 1998, 63 (27.5%) were also infected by Perkinsus sp.; the prevalence of Perkinsus sp. varied from 9.1% in January to 50% in February. To our knowledge this is the first report of Porospora sp. and Perkinsus sp. in C. chione.

[Reproducibility and accuracy of the diagnosis of cervical intraepithelial neoplasia (CIN) in 15 Italian laboratories: a national pilot project. Development of new indices of diagnostic variability. National Work Group for the expert quality control in cervico-vaginal cytopathology]. / Riproducibilità ed accuratezza nelle diagnosi di CIN in 15 laboratori italiani: progetto pilota nazionale. Messa a punto di nuovi indici clinici di variabilità diagnostica. Gruppo Nazionale di lavoro per il controllo di qualità esterno in Citopatologia cervico-vaginale.

OBJECTIVES: To assess reliability and accuracy of cervical smear diagnoses, to evaluate the effectiveness of the participation in a programme of slide exchange in increasing reliability and to re-examine the agreement in discriminating between CIN 2 and CIN 3 (merged in High grade SIL in the Bethesda System). SETTING: 15 laboratories participating on a voluntary basis throughout Italy, for a period of 1 year. METHOD: Phase one: circulation of 40 slides including all main diagnostic categories; discussion of results by representatives of participating centres. Phase two: circulation of another 40 similar slides. For each slide, not only a diagnosis but also recommendations for further examinations and a judgment on diagnostic difficulty were asked. Common measures of reliability and accuracy and (the latter only for slides on which a consensus diagnosis was reached corresponding to the histological diagnosis) were calculated; three new indices of diagnostic variability were also computed. RESULTS: Consensus diagnosis among representatives of participating laboratories on about 90% of the slides was reached both in the first and in the second phase. On 3 slides it was impossible to reach a consensus diagnosis even among external referees. In both phases, the study showed a marked variability among diagnoses, recommendations and judgment on diagnostic difficulty and, on some slides, a worrying lack of reliability in the determination of precancerous lesions. The agreement on discrimination between CIN 1 and CIN 2 was low, but it was slightly better between CIN 2 and CIN 3. No significant relationship between accuracy and workload was found. External quality control or better said, continuous quality improvement activities are essential but should be conducted in a more systematic way with greater involvement of cytotechnicians.

Reliability and accuracy of reporting cervical intraepithelial neoplasia (CIN) in 15 laboratories throughout Italy: phase 1 of a national programme of external quality control in cervical screening. The National Working Group for External Quality Control in Cervical Screening.

This paper reports results of a first phase of a pilot study to assess and improve quality of diagnoses in cervical cytological laboratories located throughout Italy. It represents the first phase of an External Quality Assurance programme (EQA). In the first phase, two sets of cervical smears representing a range of diagnoses were circulated among participating laboratories. Responses were recorded on a standardized form. Participants were asked to assess the adequacy of the smear and formulate a diagnosis. They were also asked to recommend management of the patient on the basis of the smear report and judge the degree of diagnostic difficulty of each slide. Crude index of agreement, unweighted and weighted kappas, diagnostic specific kappas, sensitivity and specificity as well as clinical indices of variability were calculated. In the second phase, two additional sets of slides were circulated after discussion of the first phase. There was striking variability between laboratories, both in terms of diagnoses offered and recommendations for management on individual slides. Assessment of the degree of difficulty of each slide was also very variable. Discrimination between CINII and CINIII was poor, confirming the choice of merging these two categories in the Bethesda classification. However, discrimination between CINI and CINII was also unsatisfactory. The results were discussed in workshops and it was possible to reach a consensus diagnosis in 35 of 40 smears. This study confirms the need for external quality control programmes.

[Spheroidal deposits of amyloid in prolactin-secreting pituitary adenomas]. / Depositi sferoidali di amiloide in adenomi ipofisari secernenti prolattina.

Nine peculiar cases of pituitary adenomas were pointed out by a retrospective investigation at the Ospedale di Legnano (from 1978 to 1984) and at the Ospedale di Circolo di Varese (from 1973 to 1986). These tumours are chromophobe adenomas with diffuse structure. Histologically they show typical, large, spheroid and concentric amyloid deposits, in addition to common, amorphous--often perivascular--ones. They were investigated by histochemical methods (Crystal-Violet, Congo-Red) and by immunohistochemical ones as well (anti-PRL and anti-GH), showing that these deposits are amyloid and are in close relation with PRL production. In particular, by immunohistochemical methods we found out that the cells of the tumours displaying spheroidal bodies do contain prolactin, not GH. The amyloid deposits are also immunohistochemically positive to anti-PRL serum, not to anti-GH serum. Finally, by considering the information present in literature, we have discussed the possible pathogenic mechanisms leading to amyloid deposits in pituitary adenomas.

Selection for necropsy: evaluation of 321 autopsied cases from 3041 consecutive deaths among residents of a defined geographical area.

By undertaking to act as 'medico necroscopo' (the medical doctors in charge of authorizing every burial) in the Unità Socio-Sanitaria Locale (USSL) 70, a region containing 92,500 inhabitants, we were able to collect over a four-year period (1985-88) information on 3371 deaths among residents of the area. Of these, 68.1% died at home, 12.6% in hospital with no autopsy, 9.5% in hospital with an autopsy and 9.8% outside of the USSL region. For the analysis, age, sex and leading cause of death were considered as explanatory variables and place of death and proportion of autopsied cases as the response variables. The factors found to be associated with deaths occurring in hospital were non-neoplastic (particularly cardiovascular and digestive) pathological conditions in patients under the age of 60. People with malignant neoplasms usually died at home, especially when they had a gastric or breast cancer. Factors associated with autopsy were: death at a young age, short length of hospitalization and death from a cardiovascular disease. Sex did not appear to play a direct role in selection for necropsy. Our results indicate that, in our and similar regions, the cases selected for autopsy form a subset that is so specific that any extrapolation to the total deceased population is misleading.

[The anatomo-pathologist as a necroscopy physician in the field: a balance-sheet of 4 years' experience]. / L'anatomo-patologo come medico necroscopo sul territorio: bilancio di un'esperienza quadriennale.

The quality of the certificates of death for 2429 cases, all the residents died at home in the USL 70 during the period 1.8.83-31.12.87, was evaluated thanks to the necroscopic activity of the Legnano's Hospital pathologist. All the information, oral or written, available about each case was used to check the family doctor's certificate. Any correction to the certificate was discussed with the certifier. During the period a gradual and progressive reduction of error in the statement of the underlying cause of death was shown. The improvement comes out using different quality indicators.

Intralaboratory reproducibility of interpretation of Pap smears: results of an experiment.

One hundred and twenty Pap-smears were examined by 3 cytotechnologists on two occasions in the Legnano Hospital Cytologic Center. A 10-category classification system was used. Chance-adjusted overall and category-specific agreement was estimated (Cohen's unweighted kappa-statistics) within and between cytotechnologists. Mean K for the Center, weighted for precision, was 0.432, S.E. 0.029 (between cytotechnologists). The most reproducible categories (between cytotechnologists) were "9: malignant cells" (K = 0.683) and "1: normal" (K = 0.533); the least reproducible categories were "4: endocervical metaplastic cells" (K = 0.024) and "8: severe squamous dysplasia CIN III" (K = 0.227).

Discovering asymptomatic non-precancerous lesions with cervical cytologic mass-screening campaigns: is it a benefit or a cost?

A total of 3357 women (88.1% married, 82.0% parous) were examined at least once in the mass-screening program carried out by the Legnano Hospital Screening Center in Nerviano (a small town near Milan with 15,600 inhabitants) in 1974, 1977 and 1980. Married women with visible portio and adequate smear were 2758: a visibly healthy cervix was found only in 1143 subjects (41.4%). A total of 1516 married women were examined at least in 2 consecutive campaigns: 1279 (84.4%) reported "no gynecologic treatment in the previous 5 years" at the first campaign, and 899 (59.3%) reported "no gynecologic treatment in the previous 3 years" at their second one. Of 350 women with "medical lesions" and 525 with "surgical lesions" recorded at the first visit, 111 (31.7%) and 232 (44.2%) reported, respectively, a "medical treatment" or a "surgical treatment" at the second visit. In women with "medical lesions" at the first visit, at the second visit a healthy cervix was found in 66.2% of the treated women and in 72.6% of the untreated patients. In those with "surgical lesions" a healthy cervix was successively found in 91.6% of treated and in 40.7% of the untreated patients.

Multifocal Purkinje-like tumor of the heart. Occurrence with other anatomic abnormalities in the atrioventricular junction of an infant with junctional tachycardia, Lown-Ganong-Levine syndrome, and sudden death.

A 13-month-old boy with intermittent preexcitation (Lown-Ganong-Levine type) and paroxysmal tachycardias of 280 to 300 beats per minute died suddenly with cardiac arrest. At necropsy the only abnormalities were in the heart. There were multifocal Purkinje cell tumors in the conduction system, including one directly at the bifurcation of the His bundle. In addition, a fault in the central fibrous body was filled with an accessory communication between the mid-portion of the atrioventricular node and both the interventricular septum and the His bundle. This accessory communication connected with the Purkinje cell tumor. Although an atrioventricular nodal bypass was thus present and could account for the preexcitation and for reentrant tachycardias, there is also the anatomic basis for an automatic arrhythmia originating within one or more of the Purkinje cell tumors, particularly the one directly within the His bundle. Dispersion of elements of the conduction system within the central fibrous body resembled the normal fetal pattern. Whether this persistence of fetal dispersion of the atrioventricular node and His bundle within the central fibrous body is causally related to the presence of multifocal Purkinje cell tumors or is only coincidence merits further consideration.

Linfoma histiocitico pleomorfico primario del bazo en un paciente con cuadro clinico de mala absorcion. / Primary pleomorphic histiocytic lymphoma of the spleen in a patient with mal-absorption syndrome

Se presenta un caso de linfoma "histiocitico" pleomorfico difuso primario del bazo, en una mujer de 76 anos de edad, con diarrea cronica y sindrome de mala absorcion. Su estudio radiologico gastrointestinal revelo engrosamiento de pliegues, dilatacion de asas y floculacion del bario y microscopicamente el intestino delgado mostro atrofia de vellosidades e infiltrado masivo de celulas plasmaticas maduras en la lamina propia, hallazgos estos compatibles con la enfermedad de cadena pesada alfa o con el llamado linfoma "mediterraneo". Se revisa la bibliografia y se hacen comentarios

Study of women who did not respond to screening for cervical cancer.

Demographic and social factors influencing the population response to cervical screening programs have been studied. Age, marital status and, to a lesser extent, place of birth and socio-economic status were the most relevant factors. On the other hand, the reasons for nonparticipation were mainly the lack of information and motivation. Personal invitations, the recall of women who did not present on the first call, and the setting up of decentralized smear collection clinics proved to be useful tools to increase attendance.