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Background: Maintaining angiogenesis inhibition and switching the chemotherapy backbone represent the current second-line therapy in patients with RAS-mutant metastatic colorectal cancer (mCRC). Regorafenib, an oral multikinase inhibitor, prolonged overall survival (OS) in the chemorefractory setting. Materials and methods: STREAM was an academic, multicenter, single-arm phase II trial, evaluating the activity of regorafenib in RAS-mutant mCRC, in terms of the rate of patients who were progression-free after 6 months from study entry (6mo-PF). Patients were pretreated with fluoropyrimidine, oxaliplatin, and bevacizumab. According to Simon's two-stage design, ≥18 patients 6mo-PF were needed in the overall population (N= 46). Secondary endpoints were safety, objective response rate (ORR), progression-free survival (PFS), and OS. Early metabolic response by [18F]2-fluoro-2-deoxy-D-glucose-positron emission tomography/computed tomography ([18F]-FDG PET/CT) scan was an exploratory endpoint. EudraCT Number: 2015-001105-13. Results: The number of patients 6mo-PF was 8/22 at the first stage and 14/46 in the overall population. The ORR was 10.9%, disease control rate was 54.6%, median (m)PFS was 3.6 months [95% confidence interval (CI) 1.9-6.7 months], mOS was 18.9 months (95% CI 10.3-35.3 months), and mPFS2 (from study entry to subsequent-line progression) was 13.3 months (95% CI 8.4-19.7 months). Long benefiter patients (>6mo-PF) significantly more often had a single metastatic site and lung-limited disease. No unexpected toxicity was reported. Grade ≥3 events occurred in 39.1% of patients, with hand-foot syndrome (13%), fatigue, and hyperbilirubinemia (6.5%) occurring mostly. Baseline metabolic assessment was associated with OS in the multivariate analysis, while early metabolic response was not associated with clinical outcomes. Conclusions: The study did not meet its primary endpoint. However, regorafenib was well tolerated and did not preclude subsequent treatments. Patients with good prognostic features (single metastatic site and lung-limited disease) reported clinical benefit with regorafenib. The exploratory metabolic analysis suggests that baseline [18F]-FDG PET/CT might be useful to select patients with a favorable outcome. A chemotherapy-free interval with regorafenib was associated with durable disease control in a selected group of patients with favorable clinical characteristics.

Pancreatic ductal adenocarcinoma (PDAC) represents a challenge for a multidisciplinary oncology team. Diagnosis of PDAC remains challenging due to overlapping imaging features with benign lesions, notwithstanding great advances with computed tomography (CT) and magnetic resonance imaging (MRI). The term "Radiomics" has recently been introduced to define a mathematical process to extract countless quantitative features from medical images (including each diagnostic technique) with high throughput computing for diagnosis and prediction. This article is an updated overview of the imaging techniques to be employed during detection and characterization of pancreatic cancer diagnostic workup. Particularly, the limitations and advantages of the different imaging techniques are discussed, with a particular focus on functional imaging. This overview is the result of a self-study without protocol and registration number. Articles published in the English language from January 2000 to January 2021 were included. We analyzed 15 papers on radiomics. The possibility of functional imaging, such as CT, MRI, and radiomics has revolutionized pancreatic imaging, improving the detection and characterization of the lesions and allowing a prognosis related to radiological features, favoring the process of personalized medicine.

Purpose Adjuvant chemotherapy improves survival of patients with gastric cancer. Intergroup Trial of Adjuvant Chemotherapy in Adenocarcinoma of the Stomach (ITACA-S) was a phase III study comparing sequential FOLFIRI followed by docetaxel/cisplatin versus 5-fluorouracil monotherapy. The intensive regimen was not superior in terms of disease-free survival (DFS) and overall survival (OS). Methods The treatment was to be started within 8 weeks from surgery. This analysis evaluates the impact of time from surgery to chemotherapy start (TSC) on outcomes. Results Out of 1,106 randomized, 1,072 patients without major violations of eligibility criteria and receiving at least one treatment cycle were analyzed. Median TSC was 50 days. Chemotherapy was interrupted in 201 (18.8%) cases, whereas it was completed without or with modifications in 277 (25.8%) and 594 (55.4%), respectively. At a median follow-up of 56.9 months, 513 progressions and 472 deaths occurred. A longer TSC was significantly associated with longer DFS (hazard ratio [HR] 0.95; 95% confidence interval [CI] 0.89-1.00; p = 0.05) and OS (HR 0.91; 95% CI 0.86-0.97; p = 0.004), after adjustment for treatment arm, age, sex, primary tumor site, number of resected nodes, and tumor stage. Better treatment compliance was associated with improved survival. Conclusions Our findings suggest that longer TSC had at least no detrimental effect on DFS and OS, whereas treatment completion had a protective effect. Our findings need to be confirmed prospectively.

Background: Some trial have demonstrated a benefit of adjuvant fluoropirimidine with or without platinum compounds compared with surgery alone. ITACA-S study was designed to evaluate whether a sequential treatment of FOLFIRI [irinotecan plus 5-fluorouracil/folinic acid (5-FU/LV)] followed by docetaxel plus cisplatin improves disease-free survival in comparison with 5-FU/LV in patients with radically resected gastric cancer. Patients and methods: Patients with resectable adenocarcinoma of the stomach or gastroesophageal junction were randomly assigned to either FOLFIRI (irinotecan 180 mg/m 2 day 1, LV 100 mg/m 2 as 2 h infusion and 5-FU 400 mg/m 2 as bolus, days 1 and 2 followed by 600 mg/m 2 /day as 22 h continuous infusion, q14 for four cycles) followed by docetaxel 75 mg/m 2 day 1, cisplatin 75 mg/m 2 day 1, q21 for three cycles (sequential arm) or De Gramont regimen (5-FU/LV arm). Results: From February 2005 to August 2009, 1106 patients were enrolled, and 1100 included in the analysis: 562 in the sequential arm and 538 in the 5-FU/LV arm. With a median follow-up of 57.4 months, 581 patients recurred or died (297 sequential arm and 284 5-FU/LV arm), and 483 died (243 and 240, respectively). No statistically significant difference was detected for both disease-free [hazard ratio (HR) 1.00; 95% confidence interval (CI): 0.85–1.17; P= 0.974] and overall survival (OS) (HR 0.98; 95% CI: 0.82–1.18; P= 0.865). Five-year disease-free and OS rates were 44.6% and 44.6%, 51.0% and 50.6% in the sequential and 5-FU/LV arm, respectively. Conclusions: A more intensive regimen failed to show any benefit in disease-free and OS versus monotherapy. Clinical trial registration: ClinicalTrials.gov Identifier: NCT01640782.

A cytogenetic analysis was performed on short-term cultures of 43 previously untreated childhood central nervous system neoplasms of various histology. The cells were obtained from pediatric patients, none of whom had received therapy before karyotypic evaluation. Successful chromosome studies were performed on 24 tumors. The most commonly detected structural abnormalities involved chromosomes 1 and 17. Other structural chromosome abnormalities involved chromosomes 3, 6, 8, 9, 11, 12, and 20.

The active site of dermatan sulfate (DS) for heparin cofactor II (HCII) was isolated in a fragment obtained by periodate oxidation, borohydride reduction, mild acid hydrolysis, and SE- and SAX-chromatography of beef mucosal and pig skin DS preparations. Characterization by mass spectrometry, one- and two-dimensional NMR spectroscopy, and HPLC analysis of disaccharides, obtained by exhaustive digestion with chondroitinase-ABC, indicates that the fragment has the prevalent structure 1, GalNAc-4SO3-[IdoA-2SO3-GalNAc-4SO3]4-R, where R is CH(CH2OH)CH(COO−)-OH. 1, is the largest DS fragment thus far isolated containing IdoA2SO3 as the only uronic acid. Its lower activity (30%) with respect to the parent polymeric DS is explainable by Tollefsen model, requiring longer polyanionic chains for formation of ternary complex with thrombin.

We have previously identified free ADP-ribose (ADPR) as a normal metabolite in mature human erythrocytes. In this study the metabolic transformations of ADPR were investigated in both supernatants from erythrocyte lysates and intact erythrocytes, loaded with ADPR by means of a procedure involving hypotonic haemolysis and isotonic resealing. In both experimental systems, the main pathway was a dinucleotide pyrophosphatase-catalysed hydrolysis to yield AMP, which was readily converted into the adenylic and inosinic nucleotide pools. To a lesser extent, ADPR underwent conversion into a compound that was identified as ADP-ribulose (ADPRu), on the basis of m.s., n.m.r. spectroscopy and enzymic analysis. ADPRu was also susceptible to degradation by the dinucleotide pyrophosphatase, which was partially purified from erythrocyte lysates and characterized with respect to its substrate specificity. Isomerization of ADPR to ADPRu was markedly enhanced by ATP. Incubation of unsealed haemoglobin-free erythrocyte membranes with labelled ADPR did not cause any transformation of this nucleotide and resulted in its trichloroacetic acid- and formic acid-resistant binding to a number of membrane cytoskeletal proteins. These proteins include spectrin, glyceraldehyde 3-phosphate dehydrogenase (Ga3PDH), three proteins of molecular masses 98, 79 and 72 kDa, which apparently comigrate with bands 3, 4.1 and 4.2 respectively, and two additional proteins of molecular masses 58 and 41 kDa. Acid-resistant binding of ADPR, as well as of NAD+, to Ga3PDH was confirmed for the enzyme purified from human erythrocytes.

Incubation of ADPribose with yeast phosphoriboisomerase resulted in the formation of an adenylic nucleotide that was identified with ADPribulose by mass spectrometry. Synthesis of [ 32 P]ADPribulose from [ 32 P]NAD + by the combined activities of commercial NAD + glycohydrolase and phosphoriboisomerase allowed us to use it as a labeled internal standard throughout the procedure of purification from trichloroacetic acid extracts of human red blood cells. ADPribulose was purified by means of three sequential reverse phase HPLC separations and its Concentration in human erythrocytes was estimated to be 0.11 ± 0.1 μM. Unsealed erythrocyte ghosts did not transform ADPribulose, which bound to specific membrane proteins with a trichloroacetic and formic acid-resistant binding. Thelabeled proteins were identified as spectrin, bands 3, 4.1, 4.2 and Glyceraldehyde 3-phosphate dehydrogenase on the basis of their relative mobilities on SDS-PAGE.

In continuous Friend leukemia cell exposure to tTAuP, the IC50 was 0.2 μM whereas in cells exposed 15 or 60 min to tTAuP followed by 72 h in drug-free medium the IC50 was 2.2 and 1.3 μM respectively. A combination of tTAuP and cisplatin (CDDP) is shown to be more active than either agent alone. Intracellular accumulation studies analysed by SXRF have shown that to achieve a cytotoxic effect, large concentrations of gold are necessary to accumulate in both the nuclear and cytoplasmic fraction. However, at cytostatic doses (1 μM), tTAuP has no effect on the cell cycle but does affect DNA synthesis. At a higher dose, greater than that necessary to induce cytotoxicity, it produces DNA damage, as observed by alkaline elution method. When cells were exposed to toxic doses of tTAuP (10 μM) in the presence of albumin, cytotoxicity was significantly reduced. Similar results were obained when cells were co-treated with L-cysteine, dithiothreitol or reduced gluthatione. Reduced cytotoxic effect can be related to the interaction with free thiol groups. According to these data it is concluded that tTAuP is highly effective in vitro; whether it is active in vivo remains to be determined.

Cell clones derived from a human melanoma metastasis selected for different integrin profiles were examined in vitro for invasive potential and biological and biochemical features potentially related to this process. Clones which expressed high levels of integrins showed high invasive potential, extracellular matrix degradation, and adhesion to gelatin-coated substrates. A correlation was also found between invasiveness and intracellular and extracellular plasminogen activator activity. Heparanase and collagenase type IV activities were apparently unrelated to invasiveness. gamma-Glutamyl transferase (GGT) activity was high in highly invasive clones, whereas melanin content was high in slightly invasive clones. Heterogeneity was also observed in cellular parameters such as cell dimensions, growth features and DNA index. The intrinsic biological and biochemical heterogeneity of a cell population derived from a single metastasis may be responsible for the different behaviour of clones, regardless of their invasive potential. Since slightly invasive cells are more differentiated than highly invasive cells, malignancy and differentiation are inversely correlated in such human melanoma clones.

An analytical procedure for the separation and determination of melphalan in human plasma was carried out. A simple high-performance liquid chromatographic method with electrochemical detection was developed taking advantage of the high sensitivity of the electrode redox reaction. The sample pretreatment consisted of a direct extraction of the interferents rather than of melphalan, owing to the difficulty of extraction of the drug, and was very simple, rapid and reproducible.

The analytical conditions in PIXE and SXRF are studied in order to optimize the detection of platinum and gold compounds in the cell and in their subcellular fractions. Calibration curves, reproducibility and detection limits were obtained with both techniques and compared. The best detection limit was achieved with SXRF. Preliminary data on the cellular pharmacokinetics of cisplatinum and gold selective distribution in nuclei were obtained by the above techniques.

The frequency of early-onset neonatal sepsis without prophylaxis is 1-5/1.000 live births. Since year '70 the most frequent causative microorganism is the group B Streptococcus (S. agalactiae, GBS), followed by Escherichia coli. The mortality rate is now reduced to 4% due to the improvement of neonatal intensive care. In the USA, the incidence of GBS early-onset neonatal sepsis has been markedly reduced by the application of the guidelines released by the Centers for Disease Control (CDC). This strategy, however, is not effective on occurrence of late-onset neonatal group B streptococcal disease. In Italy, the application of CDC guidelines is not customary, and different, often complex, protocols of obstetrical-neonatological integrated approach are applied. The frequency of infectious risk has made the GBS a paramount problem for the neonatologist, even for the legal responsibility issues resulting from the multiplicity of possible options. To reach the best level of protection of the newborn against early-onset GBS infection, the working group of providers of prenatal, obstetric, and neonatal care of the functional area of Cuneo issued an integrated protocol, in order to perform the GBS screening with the optimal culture method suggested by CDC guidelines in the highest possible number of pregnant women, and to standardize the obstetrical and neonatal management.

The two main competing models for the structure of discoidal lipoprotein A-I complexes both presume that the protein component is helical and situated around the perimeter of a lipid bilayer disc. However, the more popular "picket fence" model orients the protein helices perpendicular to the surface of the lipid bilayer, while the alternative "belt" model orients them parallel to the bilayer surface. To distinguish between these models, we have investigated the structure of human lipoprotein A-I using a novel form of polarized internal reflection infrared spectroscopy that can characterize the relative orientation of protein and lipid components in the lipoprotein complexes under native conditions. Our results verify lipid bilayer structure in the complexes and point unambiguously to the belt model.

Thyroid hormones are involved in the regulation of the GH/IGF axis. Hypothyroidism is associated with a reduction in GH pulsatility and in GH-response to stimulatory tests. In hypothyroidism, serum levels of IGF-I and IGFBP-3 fall and these changes are reserved after short-term replacement with L-T4. This study was undertaken to determine the effect of long-term replacement therapy with T4 in IGF-1 and IGFBP-3 serum levels.The study included 12 patients affected with hypothyroidism and in replacement treatment with T4. They were divided into 3 groups according to age at the beginning of treatment. Group A consisted of 4 pre-pubertal subjects with Congenital Hypothyroidism (CH) diagnosed with neonatal screening, where T4 treatment was started within 15 days of life. Group B consisted of 5 young adults where CH was clinically diagnosed at the median age of 6 months and replacement therapy started at this age. Group C consisted of 3 subjects affected with hypothyroidism secondary to thyroiditis where diagnosis and replacement treatment were delayed at age 11, 12 and 14 respectively. All subjects were matched with a control of the same age, sex, weight and pubertal stage.FT3, FT4 and TSH were in the normal range both in patients and in controls. No correlation was found between FT3 or FT4 and IGF-1 or IGFBP-3 serum levels. IGF-1 serum levels in group A (198 +/- 122 ng/ml) were significantly lower than that in group B (624 +/- 105, p0.001) and in group C (649 +/- 98, p=0.003). IGFBP-3 serum levels in group A (1.98 +/- 0.56 microgram/ml) were significantly lower than in group B (3.65 +/- 1.10, p=0.03) and in group C (4.13 +/- 0.49, p=0.003). The increase of IGF-1 and IGFBP-3 levels was seen also in control groups B and C when compared with control group A. IGF-1 and IGFBP-3 were positively correlated with age both in patients and in controls. A linear correlation was found between IGF-1 and IGFBP-3 which was positive for controls (r=0.946, p0.001) and patient group A and B (r=0.839, p = 0.005) but tended to be negative for patient group C (r=0.65, n.s.).: Our data demonstrate that long-term replacement therapy in children with hypothyroidism is associated with a physiological increase in IGF-1 and IGFBP-3. The positive correlation between IGF-1 and IGFBP-3 levels in group A and B confirm the efficacy of long-term replacement treatment on the IGF-1/BP-3 axis in pre- and post-pubertal patients treated for CH. However, this correlation tended to be negative in patients with hypothyroidism secondary to thyroiditis, suggesting that the cause of thyroid insufficiency and/or the age at the beginning of replacement therapy may have a role in the post-pubertal hormonal status in IGF-1 and IGFBP-3 balance.

Based on preclinical data, phase I/II clinical trials were performed at Istituto Oncologico Romagnolo (IOR) Operative Units (Medical Oncology Departments of Forlì, Rimini, and Ravenna, Italy) to determine the efficacy and toxicity of sequential administration of doxorubicin followed by paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) in the treatment of patients with advanced breast cancer that either had been previously untreated or that had relapsed after adjuvant therapy. In the phase I trial, 19 patients received bolus doxorubicin (50 mg/m2) followed after a 16-hour interval by paclitaxel (given at dose levels ranging from 130 to 250 mg/m2) by 3-hour infusion every 3 weeks, for a maximum of eight cycles. Paclitaxel doses were escalated in 30-mg/m2 increments if the maximum tolerated dose had not been reached in the previous dose level. Analysis of the 128 cycles assessable for toxicity demonstrated neutropenia (500/microL) in 26 courses (20.3%), with no significant clinical events. No relevant clinical cardiotoxicity was observed. The paclitaxel maximum tolerated dose was not reached at the 250-mg/m2 dose level (no grade 3 or 4 extramedullary toxicity). In the IOR phase II trial, 13 patients were treated with fixed doses of both drugs (doxorubicin 50 mg/m2 and paclitaxel 220 mg/m2). Grade 4 neutropenia occurred in 39 of the 95 cycles, but was complicated by fever in only eight cycles (8.4%); three cycles required granulocyte colony-stimulating factor support. Peripheral neurotoxicity was the most common extramedullary side effect noted. Overall clinical responses in the IOR trials included 10 complete responses (31.3%) and 15 partial responses (46.9%), with an objective response rate of 78.1%. Comparison of these results with those obtained from a phase I trial using the opposite drug sequence showed comparable overall response rates, but IOR's sequence was associated with a higher complete response rate, as well as less frequent and less severe nonhematologic toxicity.

In the present study, an HPLC-MS/MS method to confirm, in bovine urine, the most common synthetic corticosteroids illegally used as growth promoters in livestock breeding will be presented. An API III-Plus (PE-Sciex) triple quadrupole mass spectrometer, interfaced by means of an atmospheric pressure chemical ionization source to the HPLC system, was used. Urine samples were treated with a sulfatase-glucuronidase mixture to cleave the drug-conjugates and then extracted on C18 disposable columns. LC separations were performed on a reversed-phase C18 column with ammonium acetate 0.1 M/acetonitrile (60/40, v/v) as mobile phase. Detection was performed in multiple reaction monitoring mode, negative ions, selecting fragmentations characteristic of 10 corticosteroids used more frequently. Good results, in terms of sensitivity and specificity have been obtained for nine corticosteroids that can be analyzed in the same HPLC run; the limits of sensitivity achieved were 0.05-1.0 ng/ml in urine. Only a more polar corticosteroid, required a different HPLC separation. Practical applications of this technique to real samples proved that it is an effective method to confirm the illegal use of corticosteroids as growth promoter in animal. In comparison with the chemical GC-MS methods the simpler sample preparation and the faster time of analysis permit a considerable increase of sample testing per day without compromising on analytical sensitivity and specificity.

Degradation impurities have been investigated, by means of high-performance liquid chromatography (HPLC)/ mass spectrometry, in commercial ampoules of salmon calcitonin, aqueous solution, stored for up to 2 years. Oxidation derivatives of the cysteine residues in positions 1 and 7, with cleavage of the disulfide bridge, were observed in all samples. Using our reversed-phase separation, these oxidation derivatives coeluted and presented an HPLC behavior identical to the derivative of salmon calcitonin with the disulfide bridge reduced. In a previous study this last compound was tentatively identified, without confirmation by mass spectrometry, as a major degradation product of salmon calcitonin in aqueous solution.

Background. Platelet-activating factor is a mediator of inflammation involved in the blood-membrane interaction. We report that selective stimulation of complement receptors (CR1 and CR3) triggers PAF synthesis and monocyte adherence to complement-activating membranes. Methods. The synthesis of PAF was studied after stimulation of normal human adherent monocytes with F(ab)2 and Fab fragments of monoclonal antibodies specific to CR1 and CR3. CD11a, CD11b, CD18, and CD35 was studied by flow cytometry on neutrophils and monocytes. The molecular species of PAF from stimulated monocytes were identified by reverse-phase high-performance liquid chromatography coupled with mass spectrometry. Results. Anti-CR1 and anti-CR3 monoclonal antibodies induced a dose-dependent C-16 but not C-18 PAF production. The latter occurred also with monovalent Fab fragments of both anti-CR1 and anti-CR3 monoclonal antibodies, that were not internalized as seen by immunofluorescence. Adherence of monocytes to Cuprophan membranes was markedly higher (P

Pneumatically assisted electrospray was demonstrated to be a powerful ionization source for the analysis of oligosaccharides. A mass spectrometer was interfaced to an HPLC system, using this interface, to determine oligosaccharides from the enzymatic digestion of heparin separated on a reversed-phase column. To set up the technique, and particularly to clarify the ionization process, purified disaccharides, from enzymatic digestion of chondroitin sulphates, were measured. The use of a suitable counter ion in the mobile phase, tetrapropylammonium (TPA), to optimize the HPLC separation, gave, with sulphated di- and oligosaccharides, adducts [M + nTPA - (n + m)H]m-, which were unexpectedly stable to fragmentation; molecular ions [M - (n + 1)H]n-, in the presence of the counter ion, were observed only with desulphated or monosulphated disaccharides. The stability of the adducts and the use of a deuterated ion-pair reagent permitted an exact evaluation of the molecular masses of disaccharides and oligosaccharides of unknown structure. Spectra obtained in the absence of the counter ion contained singly or multiply charged molecular ions and fragmentation ions mainly from loss of the sulphate groups; under these ionization conditions the exact mass determination and interpretation of the spectra were difficult. After removal of the counter ion, tandem mass spectra could be obtained with some interesting data for the characterization of these molecules. Complete spectral analyses were performed with amounts of samples of 50 micrograms but, using microbore columns, one twentieth of this amount may give good spectra.

In continuous Friend leukemia cell exposure to tTAuP, the IC50 was 0.2 microM whereas in cells exposed 15 or 60 min to tTAuP followed by 72 h in drug-free medium the IC50 was 2.2 and 1.3 microM respectively. A combination of tTAuP and cisplatin (CDDP) is shown to be more active than either agent alone. Intracellular accumulation studies analysed by SXRF have shown that to achieve a cytotoxic effect, large concentrations of gold are necessary to accumulate in both the nuclear and cytoplasmic fraction. However, at cytostatic doses (1 microM), tTAuP has no effect on the cell cycle but does affect DNA synthesis. At a higher dose, greater than that necessary to induce cytotoxicity, it produces DNA damage, as observed by alkaline elution method. When cells were exposed to toxic doses of tTAuP (10 microM) in the presence of albumin, cytotoxicity was significantly reduced. Similar results were obtained when cells were co-treated with L-cysteine, dithiothreitol or reduced gluthatione. Reduced cytotoxic effect can be related to the interaction with free thiol groups. According to these data it is concluded that tTAuP is highly effective in vitro; whether it is active in vivo remains to be determined.

We have in this study used the 3-(4,5-dimethyl-2- thiazoyl) -2,5-diphenyl 2-H-tetrazolium bromide (MTT) end point in our histoculture drug-response assay. We have previously demonstrated that the formazan crystals formed by MTT reduction by mitochondrial succinate dehydrogenase reflect polarized light and can be measured by pixel analysis in intact tissue. The results described here indicate a total specificity of 93.8% and a total accuracy of 74.6% of the MTT end point for drug response in histoculture correlating with nine different human xenograft tumors grown in nude mice with respect to the in vivo drug response data. This in vitro system allows prediction of positive and negative responses to drugs, with a rate of 70% and 71.8%, respectively. The system described here has potential for clinical use because of the possibility of simultaneous description of the MTT values and heterogeneous response to drugs within individual tumors.

Glycosaminoglycan contents were evaluated in plasma and urine samples from volunteers treated intravenously with a mixture of dermatan sulphate and heparin, combining a novel liquid chromatographic-mass spectrometric technique for the determination of oligosaccharides from glycosaminoglycans with a classical technique for the extraction of glycosaminoglycans from biological samples (precipitation with cetylpyridinium chloride). In plasma samples dermatan sulphate and heparin can be measured for 2 h after treatment; urine excretion was detectable for 24 h. These results suggest that this novel approach is promising for future studies on the pharmacokinetics of glycosaminoglycans, although some technical aspects need further improvement, mainly regarding the procedures for sample clean-up; cetylpiridinium precipitation is a complex procedure and the recovery is limited.

The aim of this study was to assess the diagnostic sensitivity and specificity of hepatobiliary scintigraphy using a 99mTc-HIDA compound to differentiate intrahepatic cholestasis from extrahepatic forms during the first months of life. The tracer used was acid N-(2,6)-diethylacetanylido-iminodiacetic (diethyl-HIDA) with almost exclusively biliary excretion and a high concentration of radioactivity in the bile. Each neonate was injected with 0.5 mg/kg i.v. of the compound marked with a dose of 99mTc equivalent to 80-100 microCi/kg. Scintigraphic recordings were carried out every 10' for the first hour and further controls were performed at 2, 3, 4, 8 and 24 hours. Scintiphotos were obtained using a Polaroid scintillation camera, PHO Gamma V. Fifty-four patients were included in the study (34 males and 20 females) aged between 4 days and 3 months old. All patients were clinically suspected of pathologies involving the hepatobiliary tract. All cases were affected by persistent jaundice (total bilirubin between 1.8 and 39.6 mg%) with predominantly direct bilirubin (range 1.5-26.2 mg%), acholic feces and hyperchromic urine. Hepato-biliary scintigraphy revealed an intestinal excretion of tracer in 31 out of the 54 neonates examined, excluding the presence of an extrahepatic obstruction of the biliary tract. On the other hand, only 13 out of 23 cases in which no enteric excretion of the tracer was observed, was the final diagnosis one extrahepatic cholestasis. Scintigraphic tests therefore showed a 100% sensitivity associated with a specificity of only 56.52%. This demonstrates that the finding of tracer in the intestine is pathognomonic of the permeability of extrahepatic biliary ducts and that biliary atresia can be ruled out. On the contrary, the absence of the intestinal excretion of the tracer is nor constantly associated with biliary atresia. This study has confirmed this finding in 10 cases of intrahepatic cholestasis (4 hypoplasias of the intrahepatic biliary tract, 3 thick bile syndromes, 3 cases of hepatitis due to cytomegalovirus). In conclusion, the Authors state that hepato-biliary scintigraphy represents a straightforward and non-invasive diagnostic method which enables the permeability of the biliary tract to be assessed in subjects with jaundice.

The effect on DNA synthesis in cell cultures exerted by two photoactive benzocoumarins were studied. A favourable ratio between inhibition of DNA synthesis and the irradiation time periods was found in these compounds. This activity is substantially due to their ability to produce a monofunctional adduct with DNA.

We present two unrelated children, a male and a female, with signs of ectodermal dysplasia, mental retardation, agenesis/ dysgenesis of the corpus callosum, and primary hypothyroidism. Reports of ectodermal dysplasia with CNS malformations or hypothyroidism or both are rare. We suggest that the condition we describe is a distinct entity within the large group of ectodermal dysplasia syndromes and that it has a variable clinical spectrum. As both males and females are affected and in a few reports some parents show minimal signs, the inheritance is likely to be autosomal dominant.

The in vitro chemosensitivity to daunorubicin and cytosine arabinoside of blast cells derived from 35 patients affected by acute non-lymphocytic leukemia was assessed by a semiautomated tetrazolium-based colorimetric assay, by the use of p-iodonitrotetrazolium violet. The results of the in vitro testing were then compared a posteriori to clinical outcome of patients, who followed a schedule of therapy which always included the drugs tested in vitro. Three dosages of drugs were employed to allow the determination of a dose-response curve, which was obtained for all the patients. The data collected in INT assay correlated with the clinical sensitivity of patients, evaluated in terms of achievement of complete remission. For the dosage of ARA-C 500 ng/ml it was possible to establish a significant cutoff between responders and non-responders to therapy, while an acceptable distribution of sensitivity/resistance prediction was found for DNR 500 ng/ml and 5 micrograms/ml. Present results, together with rapid and easy execution of the test, encourage the use of INT assay in screening leukemic patients' sensitivity to antiblastic drugs before treatment or, in case of resistance to classical chemotherapy, in detecting individual sensitivity to alternative drugs.

Insertion reactions of sulphur dioxide with PtC σ-bonded complexes are described. These occur at about 50° in a Carius tube. The complexes obtained are of the S -sulphinate type, as indicated by their IR spectra. The S -sulphinate group shows a medium trans -effect. A comparison between carbon monoxide and sulphur dioxide insertions shows that the sulphur dioxide reactions are faster, probably because of the greater electrophilicity of sulphur dioxide.

A new series of organopalladium complexes of the type trans-[PdL2RX] (L = SeEt2, TeEt2; R = aryl group; X = halogen) containing σ-bonded organic substituents has been obtained from trans-[PdL2X2] complexes and the appropriate Grignard reagents. Analogous aryl derivatives could not be obtained from trans-PdL2X2] complexes (L = SEt2 or SPh2). The properties of the new compounds are reported and discussed. Their stabilities vary with the size of the neutral ligand and the position of bustitution on the benzene ring.

In the present study the intracellular activity of oligoadenylate synthetase and ribonuclease have been evaluated in two human breast cancer cell lines treated with tamoxifen, a well known antiestrogenic drug. Increased levels of oligoadenylate synthetase and enhanced ribonuclease activity have been found in the cultures of CG5 cell line treated with concentrations of tamoxifen inhibiting cell growth. In the experiments with the EVSA-T cell line we found neither an antiproliferative effect nor an increased oligoadenylate synthetase and ribonuclease activity. It is likely that these enzymes are involved in the mechanisms by which this drug acts as an antiproliferative agent.

Results with diethyl-IDA in the diagnosis of dilatation and obstruction of the bile ducts are reported. By comparison with methyl-IDA, the more rapid blood clearance and slower renal excretion of diethyl-IDA do not lead to interference on the part of the right kidney and ureter during the first 30' of scintiscanning. The preparation was employed in 15 subjects with persistent neonatal icterus and clinically suspected diseases of the hepatic bile ducts. Personal experience is cited for the view that, thanks to its almost exclusively biliary excretion and high bile concentration, 99mTc-labelled diethyl-IDA gives better visualisation of the bile ducts than the methyl-IDA complex.

Newborns with clinically evident forms of congenital ichthyosis are generally classified as "collodion babies" in view of the particular appearance of their skin that looks rather like a membrane of dried cellophane. This is an extremely rare clinical picture that may be the expression of various types of ichthyosis. The present paper describes two cases of collodion baby with a report on the latest discoveries about the physiopathology of the condition and the current classification of its congenital forms.

The serum concentration of protein C reactive (PCR) in preterm and term children was assessed on the first, fifth, eighth and fifteenth day of life. The data obtained show that in all cases characterised by PCR values higher than 10 mg/l on the first day of life a serious infectious pathology was present which required antibiotic treatment, monitored by subsequent PCR samplings. In all cases in which PCR was negative on the first day of life and in later samples, none had presented infectious pathologies. Analysis of the results showed that so-called borderline values (greater than 2 mg/l less than 10 mg/l) were present in some newborns at risk for the development of infectious pathologies. These children were subjected to antibiotic therapy as prophylaxis and in subsequent samples PCR became totally negative.