Relationship between Prolactin Plasma Levels and White Matter Volume in Women with Multiple Sclerosis.

BACKGROUND: The role of prolactin (PRL) on tissue injury and repair mechanisms in multiple sclerosis (MS) remains unclear. The aim of this work was to investigate the relationship between PRL plasma levels and brain damage as measured by magnetic resonance imaging (MRI). METHODS: We employed a chemiluminescence immunoassay for measuring plasma levels of PRL. We used a 1.5 T scanner to acquire images and Jim 4.0 and SIENAX software to analyse them. RESULTS: We included 106 women with relapsing remitting (RR) MS and stable disease in the last two months. There was no difference in PRL plasma levels between patients with and without gadolinium enhancement on MRI. PRL plasma levels correlated with white matter volume (WMV) (rho = 0.284, p = 0.014) but not with grey matter volume (GMV). Moreover, PRL levels predicted changes in WMV (Beta: 984, p = 0.034). CONCLUSIONS: Our data of a positive association between PRL serum levels and WMV support the role of PRL in promoting myelin repair as documented in animal models of demyelination. The lack of an increase of PRL in the presence of gadolinium enhancement, contrasts with the view considering this hormone as an immune-stimulating and detrimental factor in the inflammatory process associated with MS.

Natalizumab treatment in multiple sclerosis: the experience of S. Andrea MS Centre in Rome.

We reported a post-marketing experience of 190 patients affected by relapsing multiple sclerosis on treatment with natalizumab. Clinical findings during pre-treatment period and throughout the study were documented. Magnetic resonance imaging (MRI) scans were performed at baseline and at 6, 12, and 24 months of therapy. Cumulative proportions of patients disease activity free (i.e. absence of relapses, disability progression, MRI activity) were measured as efficacy endpoints. Despite that the baseline characteristics suggested a more severe course of disease in our sample than that of the AFFIRM trial, data on effectiveness of natalizumab were comparable. At 1 year of treatment we found 80 and 70% patients free from relapses and MRI activity, respectively, that is similar to 75 and 62% detected in the AFFIRM trial. Moreover, only 5% of our patients showed a progression of disability after a mean follow-up time of 15 months. Finally, the presence of antibodies anti-Natalizumab was strongly related to the occurrence of relapses (p = 0.002) and MRI activity (p < 0.001) even in the post-marketing experience.

The Italian version of the Unité Rhumatologique des Affections de la Main (URAM) for Dupuytren's disease: The URAM-I(10).

Introduction: Several general hand functional assessment tools for Dupuytren's disease have been reported, but none of the patient-reported-outcome measures specific to Dupuytren's disease-associated disabilities are available in the Italian language. The purpose of this study was to culturally adapt the Unité Rhumatologique des Affections de la Main (URAM) into Italian (URAM-I) and determine its measurement properties. Methods: Cross-cultural adaptation was performed according to the current guidelines. Construct validity (convergent and divergent validity) was measured by comparing the URAM-I with the Pain-Rated Wrist/Hand Evaluation (PRWHE-I), Short-Form 36 (SF-36-I) scale and finger range of motion, respectively. Factor analysis was used to investigate the URAM-I's internal structure. Reliability was assessed by internal consistency (Cronbach's alpha) and test-retest reliability by Intra-Class Correlation Coefficient (ICC). Results: This study included 96 patients (males = 85%, age = 66.8 ± 9.3). Due to the cultural adaptation, we divided the original item #1 into two separate items, thus generating the URAM-I(10). Convergent validity analysis showed a strong positive (r = 0.67), significant (p < 0.01) Pearson's correlation with the PRWHE-I. Divergent validity analysis showed a weak, negative (r < 0.3) and not significant correlation with the SF-36-I subscales, except for the physical pain subscale (r = -0.21, p < 0.05). Factor analysis revealed a 2-factor, 4-item solution that explained 76% of the total variance. The URAM-I(10) demonstrated high internal consistency (α = 0.94) and high test-retest reliability (ICC = 0.97). Conclusion: The URAM-I(10) demonstrates moderate construct validity, high internal consistency and test-retest reliability, and showed a 2-factor internal structure. Its evaluative use can be suggested for the Italian Dupuytren's population.

New insights into the photodynamics of acetylacetone: isomerization and fragmentation in low-temperature matrixes.

UV and IR photoreactivities of acetylacetone isolated at 4.3 K in four matrixes (N(2), Ne, Ar, Xe), pure and doped with O(2) are investigated, using either tunable UV and IR optical parametric oscillators, or a broad band mercury lamp. Samples are probed by UV and FTIR spectroscopies: electronic and vibrational transitions are assigned and irradiation kinetics are analyzed. Contrary to what is observed in the gas phase, stereoisomerization is the main reaction observed: UV irradiation breaks the strong H-bond of the stable enolic form of acetylacetone, leading to the observation of non-chelated forms. Isomerization among the different non-chelated forms as well as back-isomerization to the chelated form are also observed under UV irradiation. Similar reactions and reaction rates are observed for the four matrixes, indicating that the inter-system crossing to the T(1) state involved in the isomerization process is very fast, probably due to efficient coupling with phonons, in contrast with gas phase where inter-system crossing is rate-limiting. When matrixes are doped with O(2), dissociation of the non-chelated forms under UV irradiation is observed and fragments, in particular CO, are formed in large amounts. Dissociation through a Norrish type-I reaction is probably one of the reaction channels occurring during electronic relaxation: dissociation is hindered by the surrounding cage in the case of pure matrixes while fragments immediately react with O(2) in the case of doped matrixes. The differences between gas phase and cold solid medium photodynamics of acetylacetone are discussed.

Cyanobacteria from benthic mats of Antarctic lakes as a source of new bioactivities.

AIMS: To exploit the cyanobacterial diversity of microbial mats growing in the benthic environment of Antarctic lakes for the discovery of novel antibiotic and antitumour activities. METHODS AND RESULTS: In all, 51 Antarctic cyanobacteria isolated from benthic mats were cultivated in the laboratory by optimizing temperature, irradiance and mixing. Productivity was generally very low (

Silver- versus gold-catalyzed sequential oxidative cyclization of unprotected 2-alkynylanilines with oxone.

Unprecedented domino oxidative cyclization reactions of unprotected 2-alkynylanilines to give functionalized 4H-benzo[d][1,3]oxazin-4-one or benzisoxazole derivatives in moderate to good yields are achieved by silver vs. gold selective catalysis. The search for the optimal reaction conditions revealed the divergent catalytic activity of NaAuCl4·H2O and AgNO3.

Complementation of a Streptomyces lividans Leu- mutant by the Actinoplanes teichomyceticus leuC gene.

A leucine auxotroph of Streptomyces lividans (Sl), designated PC196, was unable to convert alpha-isopropylmalate into the beta-isomer. A DNA fragment from Actinoplanes teichomyceticus (At) cloned into the Streptomyces vector pIJ702 complemented PC196. Sequence analysis of the 3.0-kb insert revealed one complete ORF with high similarity to other leuC genes encoding the large subunit of isopropylmalate isomerase (IPMI), and the 5' end of a second ORF corresponding to leuD, which encodes the smaller subunit of IPMI. Further subcloning established that Sl strain PC196 is defective in the large subunit of IPMI.

Structural and functional modifications induced by ketamine on synaptosomes in the rat.

This study correlates previous observations on the effects of a general anaesthetic (2-chlorophenyl-2-methylaminocyclohexanone, trade name Ketalar), administered in vivo on the functional properties of synaptosomes from the brain of the rat, with structural modifications detected using freeze-fracture electron microscopy. The anaesthetized rats exhibited a fluidization of the synaptic membranes, as probed by stearic acid spin labels, accompanied by a decrease in the activity of acetylcholinesterase. The freeze-fracture images of synaptosomes from anaesthetized rats showed a clear statistical increase in the number of vesicles with aggregated intramembrane particles, without any significant change of the particle diameter. The hypothesis that a perturbation of the lipid protein interaction is a primary effect, in the mechanism of action of anaesthetics, is supported at present only by indirect evidence.

Postsynaptic effects of methoctramine at the mouse neuromuscular junction.

Functional studies were performed to evaluate the effects of methoctramine at the neuromuscular junction of the mouse. The presynaptic control of acetylcholine release and the postsynaptic activation of the nicotinic receptor have been analysed by means of the extracellular recording with an EPC7 Patch Clamp amplifier. This electrophysiological method revealed a dose-related inhibitory effect of methoctramine on the studied parameters. The dramatic reduction of the kinetics of the quantal conductance change indicates an action at the postsynaptic level. The effects of methoctramine have been compared with those of the muscarinic agonist oxotremorine. Concentration/response curves for the two drugs were obtained and the apparent EC50 values calculated. The effects of oxotremorine were not antagonized by 1 microM methoctramine. These findings suggest an interaction of some muscarinic agents on the postsynaptic receptor-ion-channel complex at the mouse neuromuscular junction.

Preparation of 2,5-disubstituted oxazoles from N-propargylamides.

[reaction: see text] 2,5-Disubstituted oxazoles have been prepared through the reaction of N-propargylamides with aryl iodides in the presence of Pd(2)(dba)(3), tri(2-furyl)phosphine, and NaO(t)()Bu. The reaction appears to proceed through a palladium-catalyzed coupling step followed by the in situ cyclization of the resultant coupling product.

Human IL-1 receptor antagonist from Escherichia coli: large-scale microbial growth and protein purification.

Interleukin-1 receptor antagonist (IL-1ra) is a recently discovered cytokine which specifically inhibits IL-1 pro-inflammatory activities in various experimental conditions. In this work, the growth conditions of a recombinant E. coli strain which in laboratory studies expressed human IL-1ra mostly in insoluble form, have been optimized at the level of 6-1 bioreactors and then scaled up to a 50-1 process. As a result, a high amount (0.43 g l-1 of microbial culture) of soluble, active IL-1ra has been directly obtained in the large-scale cell lysate with no need for protein solubilization. Also, an efficient purification procedure has been developed for the soluble protein, based on cation exchange expanded bed adsorption directly followed by anion exchange chromatography. This process, which does not include any intermediate dialysis step or gradient elutions, can be easily scaled up to larger production volumes and is therefore well-suited for manufacturing. As a result of the overall optimization study, more than 12 g of pure IL-1ra have been obtained from a single 50-1 fermentation run, without any denaturation/renaturation process. The final product, whose identity and purity have been checked also by MALDI-TOF and ESI-MS, shows full biological activity both in cellular assays and in in vivo experiments with Cynomolgus monkeys.

Direct and carbonylative vinylation of steroidal triflates in the presence of homogeneous palladium catalysts.

The palladium-catalyzed vinylation of several steriod 2-enyl- and 3,5-dienyl-3 triflates and estrone-3-triflate was systematically examined using vinyltributylstannane as a vinylating agent. In carbon monoxide atmosphere the insertion of a CO molecule took place and unsaturated ketones were obtained. In this way new steroid derivatives containing unsaturated side chain were produced, which can serve as starting material for further functionalization of the steroid skeleton.

Morphometric study of chromatin pattern in freeze-fractured rat liver nuclei during malignancy evolution.

Computerized image analysis was performed on freeze-fractured nuclei isolated from normal rat liver and rat liver with diethylnitrosamine induced hepatocarcinomas. Chromatin particles belonging to heterochromatin and interchromatin nuclear regions were measured, distributed into histograms and statistically processed. The ratio between nucleosome filaments (5-10 nm range diameter) and solenoid fibres (25-35 nm range) and Tukey's fitting line were calculated. The N/S ratio shows an inverse trend in treated nuclear samples with respect to the controls indicating that the malignancy progression is accompanied by characteristic derangement of the chromatin arrangement which can be evaluated by morphometric analysis.

Sampling in diagnostic morphometry: the influence of variation sources.

The variation sources relevant to a diagnostic morphometric study were analysed. The influence of each source was estimated in two experiments, performed in systems applying computer assisted interactive morphometry. In the first experiment one observer measured the areas of a large number of nuclei in a section from a grade II transitional cell carcinoma of the bladder. In the second experiment two groups of researchers, from Ancona and Kuopio, measured one field from five different samples of transitional cell tumours (including the case of grade II carcinoma). It turned out that pure interobserver variation was responsible for about a half of the total variation present in the diagnostic system. When the variation characteristics of the diagnostic system had been determined, the number of nuclei that had to be measured to reach a defined level of accuracy could be estimated. Such an estimate was also dependent on the predefined expectancy probability of reaching a correct estimate. The study showed that group morphometry (statistical, investigative morphometry) and diagnostic morphometry must be understood as two different approaches in histopathology. By applying group morphometry, good research results can be gathered with cruder measurements than in diagnostic morphometry. Because investigations in group morphometry are more standardized than in diagnostic morphometry, a larger number of structures has to be measured in diagnostic histopathology for the same level of accuracy.

Antibiotic GE37468A: a new inhibitor of bacterial protein synthesis. III. Strain and fermentation study.

GE37468A is a new thiazolyl peptide antibiotic acting on Elongation Factor Tu, produced by fermentation of Streptomyces sp. ATCC 55365. The low (< 10 mg/liter) and poorly reproducible antibiotic productivity of the original soil isolate hampered the development of a scalable fermentation process. A tenfold increase of GE37468A productivity was achieved by selection of spontaneous morphochromatic variants, which appeared at high frequency in the wild type population. The higher producing ATCC 55365/O/5 colony was tyrosinase defective and blocked in the aerial mycelium differentiation (Amy-Mel- phenotype). The fermentation process of this stable and purified strain was rapidly optimized to achieve a further fourfold improvement in a fed-batch process.

Antibiotic GE37468 A: a new inhibitor of bacterial protein synthesis. I. Isolation and characterization.

GE37468 A is a new thiazolyl peptide antibiotic obtained by fermentation of Streptomyces sp. strain ATCC 55365. It inhibits bacterial protein synthesis by acting on elongation factor Tu and is structurally and functionally related to the GE2270 class of EF-Tu inhibitors. It is active in vitro against Gram-positive bacteria and Bacteroides fragilis, and protects mice against Staphylococcus aureus infection.

Biomechanical, scanning electron microscopy, and microhardness analyses of the bone-pin interface in hydroxyapatite coated versus uncoated pins.

OBJECTIVE: To evaluate the bone-pin interface in hydroxyapatite coated versus uncoated pins. DESIGN: Eighty-four bicylindrical stainless steel external fixation pins were implanted in a test group of 14 sheep. One-half of the pins were coated with hydroxyapatite and the rest remained uncoated. INTERVENTION: Six coated pins were implanted in the left tibia of seven sheep, and six uncoated pins were implanted in the left tibia of the other seven sheep. In all sheep, the right tibia was left intact. During pin implantation, the final insertion torque was measured, and a linear external fixator was mounted on the pins. Then the medial tibial mid-diaphysis was exposed and a 5-mm resection osteotomy was performed. The sheep were killed six weeks after surgery. MAIN OUTCOME MEASURES: The extraction torque was measured on four pins removed from each sheep. Radiographic pin tract rarefaction was measured on all the pins. Two pins from each sheep were used for histologic, scanning electron microscopy (SEM), and microhardness analysis. Histomorphometric analysis was carried out on the SEM specimens at x 36 magnification. RESULTS: Radiographic pin tract rarefaction was significantly lower in the hydroxyapatite coated pins than in the uncoated pins (P < 0.001). Group average insertion torque was 960 +/- 959 N/mm in the hydroxyapatite coated pins, and 709 +/- 585 N/mm in the uncoated pins (p = not significant). Group average extraction torque was 1485 +/- 1308 N/mm and 298 +/- 373 N/mm, respectively (p = 0.0001). Histomorphometric analysis showed that the group average bone-pin contact was 50.7 +/- 16.9% in the hydroxyapatite coated pins and 27.6 +/- 7.1% in the uncoated pins (p < 0.01). Microhardness analysis showed that bone tissue close to the pins was softer than bone tissue far from them. CONCLUSION: Hydroxyapatite coating is an effective method of refining the bone-pin interface and may improve the clinical results of the external fixation technique.