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Objective. Since brain perfusion abnormalities have been described by single-photon emission computed tomography in some autoimmune diseases, the aim of the present study was to evaluate the incidence of perfusion abnormalities by brain single-photon emission computed tomography in a group of coeliac disease patients, and to investigate whether gluten intake and associated autoimmune diseases may be considered risk factors in causing cerebral impairment. Methods. Thirty-four adult coeliac patients (16 on a gluten-free diet and 18 on a gluten-containing diet, 18 (53%) with autoimmune diseases) underwent 99 m Tc -ethyl cysteinate dimer brain single-photon emission computed tomography and qualitative evaluation of brain perfusion was performed together with a semiquantitative estimation using the asymmetry index. Ten subjects on our database, matched for sex, age and ethnic group, who were proved normal by histology of jejunal mucosa (four males and six females; median age 39 years, range 27–55 years), were included as control group. Results. Twenty-four out of 34 patients (71%) showed brain single-photon emission computed tomography abnormalities confirmed by abnormal regional asymmetry index (>5%; range 5.8–18.5%). Topographic comparison of the brain areas showed that the more significant abnormalities were localised in frontal regions, and were significantly different from controls only in coeliac disease patients on unrestricted diet. The prevalence of single-photon emission computed tomography abnormalities was similar in coeliac disease patients with (74%) and without (69%) associated autoimmune disease. Conclusions. Abnormalities of brain perfusion seem common in coeliac disease. This phenomenon is similar to that previously described in other autoimmune diseases, but does not appear to be related to associated autoimmunity and, at least in the frontal region, may be improved by a gluten-free diet.

Three simple, related nucleosides, beta-L-2'-deoxycytidine (LdC), beta-Lthymidine (LdT), and beta-L-2'-deoxyadenosine (LdA), have been discovered to be potent, specific and selective inhibitors of the replication hepatitis B virus (HBV), as well as the closely related duck and woodchuck hepatitis viruses (WHV). Structure-activity relationship analysis indicates that the 3'-OH group of the beta-L-2'-deoxyribose of the beta-L-2'-deoxynucleoside confers specific anti-hepadnavirus activity. The simple nucleosides had no effect on the replication of 15 other RNA and DNA viruses, and did not inhibit human DNA polymerases (alpha, beta and gamma) or compromise mitochondrial function. The nucleosides are efficiently converted intracellularly into active triphosphate metabolites that have a long half-life. Once-daily oral administration of these compounds in the woodchuck efficacy model of chronic HBV infection reduced viral load by as much as 10(8) genome equivalents/ml serum and there was no drug-related toxicity. In addition, a decline in WHV surface antigen (WHsAg) paralleled the decrease in viral load. This class of nucleosides displays an excellent overall safety profile. The first compound, LdT, has already entered clinical trials and LdC, currently being developed as a prodrug, is expected to enter the clinic in the near future. These compounds have the potential for use in combination therapy with the goal of achieving superior viral suppression and diminishing the onset of resistance.

Worldwide, the heterosexual route is the prevalent mode of transmission of AIDS; therefore, demands have been raised for measures that block sexual spreading of the HIV infection. Development of microbicides for topical use may represent an efficacious alternative to condoms. Several approaches are being investigated. Besides surfactants, which directly act on the virus particle, and measures that enhance natural defence mechanisms, promising new candidates appear to be drugs that block the early steps of HIV multiplication. We describe herein a long-term assay which enables the establishment of whether the above drugs reversibly (virustatic action) or irreversibly (virucidal action) inhibit HIV-1 multiplication, thus allowing screening for effective and potent microbicides. We validated our assay with nucleoside (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). Following a chronic treatment, the NRTIs tested (didanosine, zalcitabine, stavudine and lamivudine) simply delayed the viral breakthrough with respect to infected, untreated controls. Under the same experimental conditions, non-nucleoside reveres transcriptase inhibitors (NNRTIs), such as MKC-442, alphaAPA, nevirapine, efavirenz and 3,4-dihydro-2-alkoxy-6-benzyl-4-oxopyrimidines (DABOs) MC 1047 and MC 1220 suppressed HIV-1 replication for the entire experimental period (40 days). When cell culture samples were evaluated for the presence of infectious virus, p24 antigen and viral DNA sequences, none of them was detected up to day 40 post-infection (p.i.). Identical results were obtained after a treatment with the above NNRTIs limited to the first 4 days p.i. Under more selective experimental conditions, that is drug treatments limited to the first 4 h p.i., nevirapine and efavirenz proved to be virustatic; in fact, viral breakthrough ensued shortly after their removal from the culture medium. Conversely, DABO MC 1220 was endowed with potent virucidal activity; in fact, at 3.5 microM it was able to suppress HIV-1 multiplication in cultures acutely infected with a very high multiplicity of infection (5 CCID50/cell), thus allowing exponential cell multiplication as in uninfected cultures for the next 40 days.

Background Bone and joint scan using 99mTc-MDP is often used in rheumatologic practice, but there is little data on its effective relevance in evaluating chronic articular inflammatory diseases. Objectives The aim of this work has been to compare bone scan results against clinical evidence of joint inflammatory involvement, evaluating with both approaches the number of affected joints (NAJ) in a group of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). Methods Bone scan was carried out in 75 consecutive patients (44 RA, 31 OA). The nuclear medicine specialist indicated in the report the list of joints showing uptake. In the same patients (without knowing the results of scintigraphy) a rheumatologist diagnosed the NAJ on the basis of clinical signs of inflammation. Then the patients were stratified in 2 * 2 tables according to the prevalence of clinical evidence or scintigraphic uptake. Results The distribution was found to be not significant (chi-square = 1.6, p = 0.2). The percentage of patients without uptake was only 5.3%. In 33 patients (44%) no clinical evidence was observed. Among these, 30 showed articular uptake (in an average of 7.0 * 4.6 joints). Considering only the patients with clinical evidence, the subjects with articular uptake were 41 out of 42 (97.6%) with an average of 8.6 * 4.6 involved joints. In these patients the ratio between number of joints with uptake and number of joints with clinical evidence (ratio uptake/clinical) was then calculated for each subject. Such a ratio resulted in an average of 2.9 * 3.6. These results were also confirmed analysing the data by sex, disease (RA or OA) and class of therapy. We then decided to assess if the greater prevalence of involved joints obtained with bone scan was correlated with an increase of inflammatory indexes. Taking into consideration the patients with clinical evidence, we performed multiple regression analysis using as dependent variable in distinct models the ratio uptake/clinical, the number of joints with clinical evidence and the number of joints showing uptake. The inflammatory indexes (ESR and CRP) were used as independent variables and age, sex, diagnosis and therapy were used as covariates. The uptake/clinical ratio did not show any significant correlation. The number of joints with clinical evidence correlates with ESR, those ones showing uptake correlates only with age, both with direct correlation. Conclusion The data obtained indicates that the articular scintigraphic uptake in AR and OA, on average, highlights a significantly higher number of joints involved as compared to the clinical test. It still remains to be defined if this is an overestimation related to the characteristics of the scan or if it is the sign of a higher sensibility in highlighting the inflammation location. An element of caution against the second hypothesis is established by the absence of correlation with the inflammatory indexes.

Molecular modeling analysis of compounds belonging to the recently published series of dihydro-alkoxy-benzyl-oxopyrimidines (DABOs), such as S-DABOs and DATNOs, gave support to the design of new 2, 6-disubstituted benzyl-DABO derivatives as highly potent and specific inhibitors of the HIV-1 reverse transcriptase (RT). To follow up on the novel DABO derivatives, we decided to investigate the effect of electron-withdrawing substituents in the benzyl unit of the S-DABO skeleton versus their anti-HIV-1 activity. Such chemical modifications impacted the inhibitory activity, especially when two halogen units were introduced at positions 2 and 6 in the phenyl portion of the benzyl group bound to C-6 of the pyrimidine ring. Various 5-alkyl-2-(alkyl(or cycloalkyl)thio)-6-(2, 6-dichloro(or 2,6-difluoro)phenylmethyl)-3, 4-dihydropyrimidin-4(3H)-ones were then synthesized and tested as anti-HIV-1 agents in both cell-based and enzyme (recombinant reverse transcriptase, rRT) assays. Among the various mono- and disubstituted phenyl derivatives, the most potent were those containing a 6-(2,6-difluorophenylmethyl) substituent (F-DABOs), which showed EC50's ranging between 40 and 90 nM and selectivity indexes up to >/=5000. An excellent correlation was found between EC50 and IC50 values which confirmed that these compounds act as inhibitors of the HIV-1 RT. The structure-activity relationships of the newly synthesized pyrimidinones are presented herein.

A series of human immunodeficiency virus (HIV) protease inhibitors, which are analogues of N-[2(R)-hydroxy-1(S)- indanyl]-5(S)-[(tert-butyloxycarbonyl)amino]-4(S)-hydroxy-6-phenyl-2-(R) - [[4-(carboxymethoxy)phenyl]methyl]hexanamide (L-694,746), a metabolite of the anti-HIV agent L-689,502, were synthesized. In these compounds, the acetic group linked to the para position of the P1' phenyl in the reference inhibitor was replaced either by the bioisosteric phosphonomethoxy group and its diisopropyl/dibenzyl derivatives, or the 1H-tetrazol-5-yl-methoxy group and its 1-benzyl derivative. In enzyme assays, phosphonomethoxy and tetrazolmethoxy analogues proved to be potent inhibitors of the HIV-1 protease, with IC50 values as low as 0.04 nM. When tested for anti-HIV-1 activity in cell-based assays, most of the new derivatives proved active, with benzyl derivatives being more active than their highly polar, unsubstituted counterparts. The dibenzylphosphonomethoxy analogue was the most active compound, with an EC50 value of 10 nM and a selectivity index of 20,000. When compounds were examined for their capability to reduce p24 levels in both acutely and chronically infected MT-4 and H9/IIIB cells, all of them were found to be active at concentrations close to those capable of preventing HIV-1-induced cytopathic effect.

Biohydrometallurgical solubilization cannotyet compete with conventional pyrometallurgical processes for recovering metals from sulphide concentrates, though the benefits deriving from its being much more environmentally friendly make it extremely attractive. Up to now, the main reason has been the relatively large size of the bioreactors and consequently the higher investment and power costs per unit mass of metal recovered compared to conventional processing. Bioreactor size is directly related to the biosolubilization process kinetics that, in turn, depend upon the suitability of the bioreactor to the process and on the prevailing biochemical conditions in the system. The Biorotor — the first reactor purpose built to match biohydrometallurgical requirements and developed by the authors - yields much faster biosolubilization kinetics than conventional bioreactors. It has been demonstrated that sulphur and iron biooxidation kinetics can be further enhanced, even in conventional bioreactors, by adding to the mineral suspension small amounts of substances capable of removing at least part of the metabolites. One of such substances is bentonite, a clay mineral that is already used as a scavenger in some commercial processes. Ferrous sulphate kinetics at least one order of magnitude higher than those reported in the literature can easily be obtained in the biorotor when the reaction takes place in the presence of bentonite.The concentration of oxidizable solids is more of a performance-limiting factor for conventional reactors than for the Biorotor and less harmful when bentonite is present in the solids suspension. Tests have been carried out in two identical Pachuca tanks operated in parallel using pure pyrite in concentrations increasing from 4.78% to 20.58% through 13.35%, one (No.I) containing a pyrite suspension and the other (No. 2) the same suspension admixed with bentonite. The ratio of pyrite solubilization rates in Pachuca No. 2 to those in Pachuca No. 1 increased almost linearly with increasing pyrite concentration. For 30% solids in Biorotor, without bentonite, the iron solubilization rate was 561 mgAdm 3 Ah -1 . Testing of Biorotor performance with bentonite confirmed the results obtained in the Pachuca, although less pronouncedly.

Acyclic glycosidopyrroles of type 3 were synthetized in good overall yields, according to the Scheme. When evaluated for antiviral activity against DNA and RNA viruses, only compound in which R1 = R2 = Ph, R3 = NH2 was found to inhibit the HIV-1 replication at concentrations that were not cytotoxic for MT-4 cells.

The synthesis and the in vitro anti-HIV-1 activity of novel pyrrolo annulated benzothiadiazepine acetic acids and some related derivatives are reported. The new compounds share chemical features with pyrrolo[1,2-d][1,4]benzodiazepin-6-one 1 and Ro 5-3335 pyrrylbenzodiazepinone 4, two inhibitors of HIV-replication at the level of reverse transcriptase (RT) and transcriptional transactivation by Tat, respectively. Two derivatives, namely methyl 10,11-dihydropyrrolo[1,2-b][1,2,5]benzothiadiazepine-11-acetic-5,5 -dioxide (5a) and 1,12b-dihydro-2H-azeto[2,1-d]pyrrolo[1,2-b][1,2,5]benzoth iadiazepin-2-one 8,8-dioxide (7a), were found to exhibit a significant, although not very potent, activity against human immunodeficiency virus Type 1 (HIV-1).

Double-phase 99mTc-methoxyisobutylisonitrile (MIBI) parathyroid scintigraphy has been proposed to detect hyperplastic parathyroid tissue, but the clinical usefulness of this technique in secondary hyperparathyroidism is still debated.Technetium-99m-MIBI parathyroid scintigraphy associated with parathyroid echography and [99mTc]pertechnetate thyroid scans were performed on 38 patients with chronic renal failure (CRF) and secondary hyperparathyroidism. In all patients, serum calcium, phosphorus, FT3, FT4, TSH, calcitonin and intact PTH (iPTH) were determined. Nine patients eventually underwent neck exploration and 28 parathyroid glands were removed.Thyroid diseases were excluded in all patients. Echography revealed parathyroid enlargement in 22/38 (58%) patients, while MIBI scintigraphy was positive in 28/38 (74%), including 5 ectopic glands. Mean serum iPTH concentration was significantly higher in MIBI-positive glands compared to MIBI-negative glands, but several discrepancies were observed in single patients. A significant positive correlation between serum iPTH and gland size was observed when MIBI-positive, but not MIBI-negative, parathyroids were considered. A paradoxical positive correlation between serum calcium and iPTH concentrations was found in MIBI-positive patients.Double-phase 99mTc-MIBI scintigraphy is positive in the majority of patients with uremic hyperparathyroidism. Comparison of scintigraphic data with morphological and functional data strongly suggests that 99mTc-MIBI scans do not reveal simple parathyroid enlargement but rather, identify the presence of hyperfunctioning (autonomous) parathyroid tissue suggestive of tertiary hyperparathyroidism.

(R)- And (S)-8-aza-9(-)[2-(phosphonomethoxy)propyl]guanine [(R)-and (S)-8-aza-PMPG] were synthesized and tested in vitro for anti-human immunodeficiency virus (HIV) activity. The synthesis of the above compounds and of (R)-9(-)[2-(phosphonomethoxy)propyl]guanine [(R)-PMPG] was carried out through the alkylation of 8-azaguanine or guanine with (R)- and (S)-2-O(-)[(diisopropylphosphono)methyl]-1-O-(tolylsulfonyl) -1,2-propanediol followed by deprotection of the phosphonic moiety. A different, even more convenient synthesis of (R)-8-aza-PMPG starting from 2-amino-6-chloro-5-nitro-4(3H)-pyrimidinone and (R)(-)[2(-)[(diisopropylphosphono)-methoxy]propyl]amine is also reported. Both (R)-8-aza-PMPG and (R)-PMPG demonstrated anti-HIV activity in the MTT assay with EC50 values of 12 and 4.5 microM, respectively. The corresponding S enantiomers were found to be less potent. When evaluated in combination with AZT, ddI, or DABO 603, (R)-8-aza-PMPG gave additive, additive, and synergistic anti-HIV-1 effects, respectively.

An automated system for the design and manufacturing of individual compensators has been implemented. The system based on computed tomography enables 3D compensation of missing tissue and tissue heterogeneities. The relationship between Hounsfield numbers and electron densities was obtained empirically. Compensator design is based on the calculation of the water equivalent thicknesses between the compensation plane and the patient surface. After calculation a styrofoam mould is cut by a computer driven machine and filled with bee's wax or tin granules. Compensator thickness is calculated by means of the conversion ratio tau, which is defined as t/x, where t is the compensator thickness equivalent to the missing tissue in the treatment geometry. Relations between tau and field size, depth of compensation plane and focus-compensation plane distance were assessed. The conversion ratio is a linear function of the missing tissue and depends markedly on field size; for a 10-cm-deep compensation plane at 1 m from the accelerator target the tau value, calculated for bee's wax, decreases by 25% from 7 x 7 cm2 to 23 x 23 cm2 field size. Conversion ratio rises by approximately 10% for a 3-cm increase in compensation plane depth and reduces by about 5% when increasing the focus-compensation plane distance from 100 cm to 140 cm. It must be stressed that a 10% variation of tau, for bee's wax, involves only a 2% dose variation in the compensation plane. Therefore, for compensator design it is enough to consider tau as depending on field size only. Compensation effectiveness has been tested by a film-densitometric technique using phantoms with tilted incident surfaces and heterogeneities. The results show that the compensators reduce the flatness of the beam profile below 4% and increase the relative dose uniformity on the compensation plane from 18% to 60%.

As an approach to more extensive structural modifications of [(oxazolylphenoxy)alkyl]isoxazoles, we synthesized new compounds characterized by the replacement of the isoxazole nucleus with furan, pyrrole, and thiophene rings and by the presence of a ketocarbonyl group in the aliphatic chain connecting these pentatomic heterocycles to the 4-(4,5-dihydro-2-oxazolyl)phenoxy, 4-(ethoxycarbonyl)phenoxy, and 4-carboxyphenoxy moieties. Some pentamethylene derivatives were also prepared, and their antirhinovirus activity was compared to that of the corresponding ketomethylene derivatives. Syntheses were carried out by Friedel-Crafts acylation of the above pentatomic heterocycles and subsequent reaction of chloroalkyl ketones with the proper 4-substituted phenol. Reduction of the ketone function afforded the related polymethylene derivatives. The new compounds were tested for antirhinovirus activity and cytotoxicity in comparison with WIN 51711, used as reference drug. Inspection of the structure-activity relationships revealed that the thiophene ring and the carbonyl group are the structural components which to a large extent contribute to the positive biological profile in terms of both wideness of spectrum and low cytotoxicity. Among the various derivatives, compounds 8e,d showed in vitro the same potency of WIN 51711 but a cytotoxicity at least 10 times lower.

Some aza and deaza analogues of the anti-HIV agent 2',3'-dideoxy-3'-oxoadenosine (isoddA) (8-aza-, 8-aza-1-deaza, 8-aza-3-deaza-, 1-deaza-, and 3-deaza-isoddA) were synthesized and found inactive against HIV in vitro. The hypothesis that the inactivity of these isonucleosides might be due to their poor affinity for cellular nucleoside kinases was checked by the synthesis of a series of 5'-[bis(2,2,2-trichloroethyl) phosphate] triesters and 5'-phenyl phosphoramidate derivatives which, acting as membrane soluble prodrugs, could release the free phosphate form inside the cell. The 5'-(phenylmethoxy)alaninyl phosphate derived from 8-aza-isoddA was found active against HIV-1 and HIV-2 with a potency similar to that of isoddA, while the anti-HIV potency of 5'-(phenylmethoxy)alaninyl phosphate of isoddA proved remarkably higher than that of isoddA, in particular against HIV-2, being similar to that of AZT. Further evidence that 8-aza-isoddA could behave as anti-HIV agent, provided that it is activated as phosphate, was obtained by the synthesis of its 5'-triphosphate derivative, which proved to be an active inhibitor of HIV-1 recombinant reverse transcriptase.

Novel 3,4-dihydro-6-benzyl-4-oxopyrimidines (DABOs), variously substituted at both the C-2 and C-5 positions of the pyrimidine ring, proved to be specific inhibitors of the human immunodeficiency virus type 1 (HIV-1) in vitro. Some compounds showed potency at micromolar doses, no cytotoxicity at the maximum testable doses and selectivity indexes comparable to that of 2'-3'-dideoxyinosine (ddI). Mode of action studies suggested that DABOs interfered with a step of the virus multiplication cycle following adsorption and preceding integration. Enzyme assays indicated that DABOs targeted HIV-1 reverse transcriptase: they inhibited the RNA-dependent DNA polymerase activity in a template-dependent manner and, to a lesser extent, the DNA-dependent DNA polymerase activity. No inhibition of the RNase-H associated activity was observed. When DABOs were assayed in combination with 3'-azido-3'-dideoxythymidine (AZT) or ddI against HIV-1 in cell cultures, a slightly synergistic inhibitory effect was observed. The combination of DABO 546 and AZTTP in enzyme assays showed that the two compounds were kinetically mutually exclusive.

Earlier investigations on laboratory and pilot plant scale have shown the resort to microorganisms to be a practicable approach to the problem of purifying mineral dressing plant reject waters from residual flotation reagents and/or metal ions. In spite of the proven effectiveness of this method, one major drawback, namely the pathogenicity of some microorganisms, has so far hampered its application on a commercial scale. A research programme, aimed at developing a microbial reject water purification technique utilizing non-pathogenic strains was thus drawn up and is currently being implemented. Strains such as Bacillus subtilis, Saccharomyces cerevisiiae, Bifidobacterium bifidum, Lactobacillus acidophilus and Acinetobacter calcoaceticus , which are not harmful to human health, some of them being commonly found in the human intestine, have been successfully tested for removing alkylsulphates, alkylamines and fany acids from solutions simulating flotation plant tailings waters. Removals as high as 90% in less than 48 h can be easily achieved with no nutrient requirements, since in most cases the flotation reagent residue to be removed is metabolized by the microorganisms themselves.

The authors report on their experience with liver hemangioma (LH) diagnosis by means of a simplified method--that is, the simultaneous, in vivo, double labelling of liver reticuloendothelial system (RES) and of red blood cells (RBC) by 99mTc. Twenty-eight patients with US diagnosis of suspected LH and 15 controls were examined after sequential iv injection of SnCl2, of 99mTc-mucolloid albumin and, after liver scintigraphy, of 99mTc-pertechnetate to conclude in vivo RBC labelling. All patients underwent CT and, if necessary, CT-guided biopsy. Focal colloid defects filled after RBC labelling were shown in 20/22 patients with unquestionable LH. No colloid defects were shown in 6/28 cases (expansive process). 15/15 controls showed unchanged non-filling defects after double labelling. Finally, the authors point out that, in the diagnosis of LH, sequential double labelling of liver RES and RBC appears to be a quicker scintigraphic technique than conventional ones. Moreover, this technique has the same high specificity and sensitivity as more time-consuming ones.

21184 Background: The prognostic value of flow cytometry DNA ploidy and Ki-67 expression in colorectal carcinomas has not been defined yet. The study tries to correlate tumoral DNA ploidy and proliferative activity (Ki-67) with therapy response, Overall Survival (OS), Disesase Specific Survival (DSS) and Disease Free Survival (DFS). Methods: From 01/09/02 to 30/06/05, 3 samples of cancer tissue and 1 sample of normal mucosa has been collected from all operating pieces of colorectal cancer. These samples were frosen and later disgregated, treated and coloured with Propidium Iodide. DNA ploidy was evaluated by FACSCalibur cytofluorimetry. Normal mucosa tissue was our internal control. Ki-67 was evaluated by immuno-histochemistry in all tumoral samples. All the patients were cured with chemo- and/or radiotherapy in our divisions. Results: 67 patients (M/F 35/32); median age was 70; staging: 19% I, 33% II, 30% III, 18% IV. We found aneuploidy in 65,7% of carcinoma and Ki-67 median expression was 55%. DNA tumoral heterogeneity was present in 27% of patients. DNA aneuploidy correlates with advanced disease stage at diagnosis (p55%) do not show evident disease versus 100% of patients with DNA diploidy and lower Ki-67. Tumoral aneuploidy correlates in a significative way with lower OS (48% vs 89% of diploid patients), lower DSS (tumor death happened just in patients with aneuploid DNA in every disease stages), with lower DFS (18% vs 86% of diploid patients). Univariated analysis stated that aneuploidy determinates an Odd Ratio=5,7 to develop disease progression (p=0,033). At the moment Ki-67 expression with 55% cut-off does not seem to correlate with OS, DSS and DFS. Conclusions: Preliminar results (the study is still in progress) seem to suggest that cytofluorimetric DNA-ploidy has a prognostic and predictive significance in colo-rectal carcinomas. Ki-67 expression (immuno-histochemistry) has an uncertain significance. The small number of patients and the short follow-up do not allow us to reach any definitive conclusions, but the study is worth to go on. No significant financial relationships to disclose.

The globin CACCC boxes are absolutely required for the appropriate regulation of the β-like globin genes. While the β-globin CACCC box binds EKLF/KLF1, a likely adult switching factor, analogous factors, interacting with the γ-globin gene and predicted to regulate the fetal stage of hemoglobin switching, have so far been elusive. By using yeast one hybrid assay, we have isolated four KLFs, KLF1, 2, 4, and 6, that bound the γ-CACCC bait. To establish their role in globin regulation and in the switching of hemoglobins, these factors were compared to four other KLFs already established or putative globin regulators, KLF3, 11, 13 and 16, mainly evaluating their ability to bind and transactivate the ε-, γ- and β-globin gene. γ-CACCC binding at variable intensities was confirmed in band shift assay for all four isolated KLFs, for KLF3 and, faintly, for KLF13. The ε- and β-CACCC were bound by the same factors with similar affinities with the exception of KLF3 and KLF13 that bound stronger to the β- and ε- than to the γ-CACCC box. On the other hand, KLF11 and 16 did not produce any specific complex in band shift assays with anyone of the globin CACCC boxes. More relevant differences were observed among the factors in the transactivation of single and dual luciferase reporters in both K562 and MEL cells. In these assays, most factors presented peculiar modulatory properties and specific promoter tropism. Several factors presented bidirectional activity displaying in the same time the capacity to stimulate and repress different globin promoters. KLF1 and 4 were the strongest stimulators of the β-globin promoter in both cell lines, whereas KLF2 activated the β-promoter only in K562 cells. KLF1 and especially KLF4 consistently repressed ε-globin expression especially in MEL cells. KLF3 behaved always as a general globin repressor in MEL cells, but acted as a weak stimulator of the γ- and ε-promoter in K562 cells. KLF4 was the strongest inhibitor of the ε-globin gene. KLF13 significantly stimulated the γ-promoter in both cell lines, whereas KLF3, 4 and 6 showed statistically significant stimulation only in MEL cells. By RT-PCR analysis we found that KLFs were highly variable in their tissue expression and that KLF1, 3 and 13 had the highest expression in erythroid tissues. Thus the level of tissue expression should ultimately determine which factors are really active in physiological conditions. Taken together our binding and expression studies suggest that several KLFs have the potential to modulate the activity of the globin genes and that the resulting globin expression will depend on the vectorial sum of the relative activities of the factors expressed at any given time of development. Furthermore, as some KLFs, like KLF1 and 4, exert opposite effects on fetal and adult globin genes, their role in hemoglobin switching may be direct and not only dependent on their ability to mediate promoter competition for the LCR.

Capacité de charge et territoire — 2. Conséquences spatiales du manque de planification globale — 3. Tourisme et occupation des sites — 4. Dommages écologiques dus à l'ouverture d'espaces au flux touristique — 5. Planification du développement touristique — 6. Conclusions.

The plasma cholic acid, chenodesoxycholic acid and desoxycholic acid levels were studied by spectrofluoremetry in 153 cases. The values of 67 controls with no evidence of hepatobiliary or intestinal disease were compared with those of 86 patients with liver and biliary tract disease. The fasting values failed to provide more diagnostic information than did conventional laboratory assays. Plasma bile acid concentrations exceeding 2.5 mu mol/l are conclusive of liver or biliary disease. A cholic acid/chenodesoxycholic acid quotient higher than 1.0 is a sign of cholestasis. Estimation of bile acids after food intake was found more informative. The plasma cholic acid- and chenodesoxycholic acid levels underwent a considerable increase 1 to 2 hours after meals. A more marked increase of chenodesoxycholic acid than of cholic acid (the ratio of the two being in excess of 1.0) is indicative of cholestasis and is most marked in primary biliary cirrhosis.

Helicobacter pylori since Marshall and Warren's discovery has been an object of interest of gastroenterologists and many researchers of other specialties. What needs to be highlighted is also the growing interest of dentists in the role of oral residue of H. pylori in oral pathologies such as burning mouth syndrome, periodontitis and gingivitis. With the development of medical techniques more studies using highly specific diagnostic methods are performed in order to determine the transmission pattern of bacterial infection. Suggested faecal-oral and oral-oral routes of bacterial transmission raised interest in molecular biology methods as tools for the study of these environments. Additionally, co-existence of helical and coccoidal forms of H. pylori in the mentioned niches raised the question whether the latter is potentially pathogenic. This is why molecular biology is now giving a great opportunity to explore parts of the human body that could not have been diagnosed before using only gold standard diagnostic methods. Molecular techniques have shown their usefulness in examining the potential virulence of coccoid forms of bacterium. This review was created also to summarize the knowledge about molecular methods, especially different PCR techniques, as diagnostic tools that can help medical teams during regular diagnosis of gastritis.

The recent discovery of gold finely intergrown with the complex metal sulphides in the Tertiary volcanites of the Serrenti-Furtei district in central and southern Sardinia, prompted the investigation of gold extraction methods that are both economically viable and environmentally friendly. Here two types of integrated processes for gold extraction are compared: roasting plus cyanidation and bioleaching plus cyanidation. With the first process, overall gold recoveries as high as 85% are achieved, whereas with a variant of the second, total gold recovered does not exceed 77%. Both types of processes investigated are described in detail. The findings of the investigation suggest that the advantages offered by the biohydrometallurgical processes—especially from the environmental viewpoint—may compensate the lower gold recovery they yield.

Differenti studi hanno riscontrato nei pazienti con diagnosi di disturbo psicotico una diffusa compromissione delle funzioni esecutive, in particolare della Working Memory, e una tendenza a prendere decisioni rapidamente dovuta ad errori nel processamento di raccolta delle informazioni presenti nel contesto (Jumping To Conclusion, JTC). Obiettivo del presente studio é quello indagare la presenza di una possibile correlazione tra Working Memory e Jumping To Conclusions in un campione di pazienti al primo episodio psicotico. Per il presente studio sono stati valutati 41 pazienti all’esordio psicotico (58,5% M), di età media 29,63 (DS=10,285) e 89 controlli sani (47,2% M), di età media 33,31 (DS=12,817). I dati hanno mostrato che il Jumping To Conclusions è presente in 45 degli 89 soggetti del gruppo di controllo (51,1%) e in 31 dei 38 individui del gruppo dei casi (81,6%). Dalle percentuali ricavate, si nota come il gruppo dei casi fosse più propenso a “saltare alle conclusioni” rispetto ai controlli (χ2=10,28; p=0,001). Per quanto riguarda i compiti di Working Memory, i pazienti psicotici hanno ottenuto punteggi peggiori rispetto ai controlli. Infine, le analisi di correlazione svolte all’interno del gruppo dei casi rilevano la presenza di una correlazione significativamente positiva tra il punteggio ottenuto al Beads Task e tutti i compiti di Working Memory: Digit Span (rho=0,28; p=0,003), Digit Symbol (rho=0,27; p=0,002), Arithmetic (rho=0,26; p=0,003) e Block Design (rho=0,29; p=0,001). I risultati esposti sembrano inoltre supportare, in accordo con la letteratura, l’ipotesi di una possibile correlazione tra alterazioni a carico della Working Memory e deficit di raccolta dati Jumping To Conclusions.

With the aim of developing novel inhibitors of human immunodeficiency virus, various derivatives (10-17) related to 5H-pyrrolo[1,2-b] [1,2,5]benzothiadiazepine (PBTD) were prepared and tested in vitro. The title tricyclic derivatives were obtained by intramolecular cyclization of the open-chain intermediate arylpyrrylsulfones, followed by N-alkylation at position 10. Among test derivatives some 10-alkyl-5H-pyrrolo[1,2-b] [1,2,5]benzothiadiazepin-11(10H)-one-5,5-dioxides were found to exert potent and specific activity against HIV-1. In particular, 7-chloro derivatives 11i and j showed a potency comparable to that of nevirapine. However, when the chloro atom was shifted to the 8 position, the related products were scarcely active or totally inactive. Replacement of the pyrrole with pyrrolidine led to inactive products and the reduction of SO2 to S strongly diminished the antiviral potency. PBTD derivatives active in cell cultures were also inhibitory to the recombinant HIV-1 RT in enzyme assays, thus allowing the conclusion that PBTDs are a new class of non-nucleoside reverse transcriptase inhibitors (NNRTIs).

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