Production and purification of higher molecular weight chondroitin by metabolically engineered Escherichia coli K4 strains.

The capsular polysaccharide obtained from Escherichia coli K4 is a glycosaminoglycan-like molecule, similar to chondroitin sulphate, that has established applications in the biomedical field. Recent efforts focused on the development of strategies to increase K4 polysaccharide fermentation titers up to technologically attractive levels, but an aspect that has not been investigated so far, is how changes in the molecular machinery that produces this biopolymer affect its molecular weight. In this work, we took advantage of recombinant E. coli K4 strains that overproduce capsular polysaccharide, to study whether the inferred pathway modifications also influenced the size of the produced polymer. Fed-batch fermentations were performed up to the 22 L scale, in potentially industrially applicable conditions, and a purification protocol that allows in particular the recovery of high molecular weight unsulphated chondroitin, was developed next. This approach allowed to determine the molecular weight of the purified polysaccharide, demonstrating that kfoF overexpression increased polymer size up to 133 kDa. Higher polysaccharide titers and size were also correlated to increased concentrations of UDP-GlcA and decreased concentrations of UDP-GalNAc during growth. These results are interesting also in view of novel potential applications of higher molecular weight chondroitin and chondroitin sulphate in the biomedical field.

Acute aortic dissection in the young: clinical series.

Acute aortic dissection (AAD) is an uncommon lethal cardiovascular emergency demanding prompt diagnosis and aggressive therapeutic intervention. Although it usually affects males over 60 years of age, it may also occur in young adults with specific risk factors such as Marfan syndrome, bicuspid aortic valve and larger aortic dimensions. Moreover, it should be underlined that it is frequently associated with unusual presentation and that the mortality risk is similar to older AAD patients. Thus ''a call to arms'' of the medical community is needed to better understand the spectrum of acute aortic syndromes and to define appropriate diagnostic and therapeutic pathways. We report a series of 6 young patients with type A - AAD referred over the past years at our institute.

Neuropsychological assessment in HTLV-1 infection: a comparative study among TSP/HAM, asymptomatic carriers, and healthy controls.

BACKGROUND: Human T cell lymphotropic virus type 1 (HTLV-I) can cause tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM) and adult T cell leukaemia/lymphoma. More recently other diseases such as isolated peripheral polyneuropathy, myopathy, artropathy, and uveitis have been associated with this retrovirus. Only a few uncontrolled studies, without necessary exclusion criteria, have described mild cognitive deficits among TSP/HAM patients. To further clarify this the authors evaluated, through neuropsychological testing patients with TSP/HAM and asymptomatic infected carriers, comparing both groups with healthy controls. OBJECTIVES: To verify the presence of cognitive deficits among TSP/HAM patients and asymptomatic HTLV-1 infected carriers. In addition, the authors aimed to investigate if these deficits correlated with the degree of motor impairment in TSP/HAM patients. METHODS: From a cohort of 501 HTLV-1 infected people the authors selected, according to predefined inclusion and exclusion criteria, 40 asymptomatic HTLV-1 carriers and 37 TSP/HAM patients. Neuropsychological testing was blindly performed in both groups and their scores were compared with those obtained from controls. RESULTS: Both the HTLV-1 carrier group and the group of patients with TSP/HAM exhibited a lower performance in neuropsychological tests when compared with controls. Asymptomatic infected carriers and TSP/HAM patients did not differ in their cognitive results. Also, there was no relation between the degree of motor disability and cognitive deficits in the TSP/HAM group. Psychomotor slowing and deficits in the some domains characterised the neuropsychological impairment in HTLV-1 infection: verbal and visual memory, attention and visuomotor abilities. CONCLUSIONS: TSP/HAM as well as asymptomatic infection can be associated with mild cognitive deficits. This finding, if confirmed by further studies, will permit the inclusion of cognitive impairment among the neurological manifestations of HTLV-1.

[Epidemiology of accidents related to sea-swimming in the Tuscany Region using a health-promotion strategy. Preliminary report]. / Epidemiologia degli incidenti correlati all'attività balneare nella Regione Toscana per una strategia di promozione della salute--nota preliminare.

In the summer season 1999 an integrated epidemiological surveillance system (involving mobile emergency medical services, first aid and tourist stations, hyperbaric medical centres, bathing attendants) of sea-bathing-related accidents was set up on the coasts of Tuscany, central Italy, aimed at health promotion and education. The pilot phase allowed to collect a first set of information on periods and time with highest incidence of events, type of assistance delivered, kind of accident (trauma or illness) and seriousness of the event as codified by emergency medical services. The pilot experience also pointed out the changes to detection tools needed in order to obtain more precise and comparable data. Such corrections, introduced during the summer season 2000, could contribute to the creation of a model with potential applications in other Italian and European coastal regions.

Static magnetic fields for treatment of fibromyalgia: a randomized controlled trial.

OBJECTIVE: To test effectiveness of static magnetic fields of two different configurations, produced by magnetic sleep pads, as adjunctive therapies in decreasing patient pain perception and improving functional status in individuals with fibromyalgia. DESIGN: Randomized, placebo-controlled, 6-month trial conducted from November 1997 through December 1998. SETTING AND SUBJECTS: Adults who met the 1990 American College of Rheumatology criteria for fibromyalgia were recruited through clinical referral and media announcements and evaluated at a university-based clinic. INTERVENTIONS: Subjects in Functional Pad A group used a pad for 6 months that provided whole-body exposure to a low, uniform static magnetic field of negative polarity. Subjects in the Functional Pad B group used a pad for 6 months that exposed them to a low static magnetic field that varied spatially and in polarity. Subjects in two Sham groups used pads that were identical in appearance and texture to the functional pads but contained inactive magnets; these groups were combined for analysis. Subjects in the Usual Care group continued with their established treatment regimens. OUTCOME MEASURES: Primary outcomes were the change scores at 6 months in the following measures: functional status (Fibromyalgia Impact Questionnaire), pain intensity ratings, tender point count, and a tender point pain intensity score. RESULTS: There was a significant difference among groups in pain intensity ratings (p = 0.03), with Functional Pad A group showing the greatest reduction from baseline at 6 months. All four groups showed a decline in number of tender points, but differences among the groups were not significant (p = 0.72). The functional pad groups showed the largest decline in total tender point pain intensity, but overall differences were not significant (p = 0.25). Improvement in functional status was greatest in the functional pad groups, but differences among groups were not significant (p = 0.23). CONCLUSIONS: Although the functional pad groups showed improvements in functional status, pain intensity level, tender point count, and tender point intensity after 6 months of treatment, with the exception of pain intensity level these improvements did not differ significantly from changes in the Sham group or in the Usual Care group.

Responsiveness of fibromyalgia clinical trial outcome measures.

OBJECTIVE: To assess the responsiveness of the Fibromyalgia Impact Questionnaire (FIQ), patient ratings of pain intensity, number of tender points, and total tender point pain intensity score to perceived changes in clinical status in patients with fibromyalgia (FM). METHODS: Using data from a randomized placebo controlled study evaluating efficacy of magnetic therapy in patients with FM, the ability of primary outcomes to detect clinically meaningful changes over a 6 month period was assessed by: (1) degree of association between outcome change scores and patient global ratings of symptom change (Spearman rank-order correlations); (2) ability of these scores to discriminate among groups of patients whose perceived health status had changed to varying degrees (ANOVA); (3) ability of these scores, individually and jointly, to discriminate between patients who had reported improvement and those who did not (logistic regression); (4) effect size, standardized response mean, and Guyatt's statistic were calculated to quantify responsiveness. RESULTS: Correlations showed the outcome measures were moderately responsive to perceived symptomatic change. For FIQ, pain intensity ratings and number of tender points, differences in change scores between globally improved and unchanged groups and between globally improved and worsened groups were significant; for total tender point pain intensity, the globally improved differed from worsened group. FIQ outperformed the other measures in discriminating between patients who reported improvement from those who did not. Summary statistics were consistent with discriminatory analyses, indicating the measures were sensitive to improvement, but relatively unresponsive to decline. CONCLUSION: The FIQ was the most responsive measure to perceived clinical improvement and we recommend its inclusion as a primary endpoint in FM clinical trials.

Prognostic value of triple phase bone scanning for reflex sympathetic dystrophy in hemiplegia.

Twenty-two patients with cerebral vascular accident (CVA), clinically confirmed by head computed tomography, were observed for symptoms of the reflex sympathetic dystrophy syndrome (RSDS). All patients received triple phase bone scans; 16 scans were positive for RSDS. Patients with negative scans had no symptoms of RSDS. Five patients with positive scans had RSDS symptoms at the time of bone scanning. Seven of 11 patients with positive scans but no symptoms of RSDS at the time of bone scan developed symptoms of RSDS within six months. We found a significant relationship between positive bone scans and the subsequent development of RSDS (p < 0.01). Considering only those patients who were asymptomatic for RSDS at the time of bone scanning, we found bone scanning to be a good predictor for the future development of clinical RSDS. We found the correlation between positive bone scans and the subsequent development of clinical RSDS in previously asymptomatic individuals to be statistically significant (p < 0.05). We conclude that bone scans may be a good predictor of patients at risk for developing clinical RSDS after CVA.

Photolysis-laser-induced fluorescence diagnostic for GaCI.

A spatially resolved optical diagnostic for GaCl is described. The technique uses 248-nm excimer laser radiation to photolyze GaCl and also to excite fluorescence in the resulting gallium atoms. This yields gallium fluorescence at 245, 266, 287, 294, 403, and 417 nm. The method is specific to GaCl(3) and is not affected by the presence of a GaCl3 precursor. Gallium fluorescence is linear in both the GaCl partial pressure and the laser energy under specified conditions. The photophysics of this process are contrasted to related GACl studies of dissociative excitation at 193 nm.

The comparative concurrent validities of the Shipley Institute of Living Scale and the Henmon-Nelson Tests of Mental Ability.

This study compared the abilities of the Shipley Institute of Living Scale and the Henmon-Nelson Tests of Mental Ability to predict Wechsler Adult Intelligence Scale-Revised (WAIS-R) scores in psychiatric hospital patients. The Henmon-Nelson DIQs accounted for about 50% more WAIS-R Verbal and Full Scale IQ variance than did the Shipley IQs, apparently because of their higher correlations with the Information, Vocabulary, and, perhaps, Similarities subtests. Because Henmon-Nelson scores were more variable and generally higher than their WAIS-R counterparts, statistical adjustments were needed to optimize Wechsler IQ estimates. Therefore, regression formulae and a conversion table for the estimation of WAIS-R Full Scale IQs from Henmon-Nelson and Shipley intelligence scores also are presented.

Spectroscopic characteristics and site I/site II classification of cis and trans benzo[a]pyrene diolepoxide enantiomer-guanosine adducts in oligonucleotides and polynucleotides.

The highly tumorigenic isomer (+)-7,8-dihydroxy-anti-9, 10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+)-anti-BPDE] and its non-tumorigenic enantiomer (-)-anti-BPDE are known to react predominantly with the exocyclic amino group (N2) of deoxyguanine in DNA and to form adducts of different conformations. The spectroscopic characteristics (UV absorbance, fluorescence and circular dichroism) of stereochemically defined (+)-trans, (-)-trans, (+)-cis and (-)-cis d(5'-CACATGBPDETACAC) adducts in the single-stranded form, or complexed with the complementary strand d(5'-GTGTACATGTG) in aqueous solution, were investigated. The spectroscopic characteristics of the double-stranded d(5'-CACATGBPDETACAC).d(5'-GTGTACATGTG) adducts can be interpreted in terms of two types of conformations. In site I-type conformations, there is an approximately 10 nm red shift in the absorption maxima, which is attributed to significant pyrenyl residue-base interactions; in site II-type adducts, the red shift is only approximately 2-3 nm, and the pyrene ring system is located at external, solvent-exposed binding sites. The spectroscopic characteristics of the BPDE-modified duplexes are of the site II type for the (+)- and (-)-trans, and of the site I type for the (+)- and (-)-cis adducts. In adducts derived from the binding of (+)-anti-BPDE to poly(dG-dC).(dG-dC) and poly(dG).(dC), the trans/cis BPDE-N2-dG adduct ratio is 6 +/- 1; in the case of (-)-anti-BPDE this ratio is only 0.4 +/- 0.1 and 0.6 +/- 0.15 in poly(dG-dC).(dG-dC) and poly(dG).(dC) respectively. The spectroscopic properties of these BPDE-modified polynucleotide adducts are consistent with those of the BPDE-modified oligonucleotide complexes; the cis adducts are correlated with site I adduct conformations, while the trans adducts are of the site II type. The correlations between adduct characteristics and biological activities of the two BPDE enantiomers are discussed.

Dependence of conformations of benzo[a]pyrene diol epoxide-DNA adducts derived from stereoisomers of different tumorigenicities on base sequence.

The conformations of covalent adducts derived from the binding of the highly tumorigenic stereoisomer (+)-trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyren e [(+)-anti-BPDE] and its nontumorigenic (-)-anti-BPDE isomer with poly[(dG).(dC)], poly[(dG-dC).(dG-dC)], poly[(dT-dC).(dG-dA)], and poly[(dA-dC).(dG-dT)] were investigated by employing UV absorbance and linear dichroism methods. The degrees of orientation of the BPDE residues (bound covalently to N2 of deoxyguanosine), relative to the DNA bases, are most pronounced in the alternating and nonalternating (dG).(dC) polymers and decrease in polymers with neighboring dA.dT base pairs. The tumorigenic (+)-anti-BPDE isomer gives rise predominantly to external (solvent-exposed) site II adducts, while the (-)-enantiomer gives rise predominantly to site I adducts with significant carcinogen-nucleoside interactions. In the mixed (dA-dC).(dG-dT) and (dT-dC).(dG-dA) copolymers, the (+)-anti-BPDE isomer also binds predominantly to N2 of deoxyguanosine, but the adducts are weakly oriented with respect to the DNA bases. The incidence of site II adducts is considerably reduced as compared to the (dG).(dC) and (dG-dC).(dG-dC) polymers, and there is a greater proportion of site I adducts; the presence of a significant proportion of unordered adduct forms is also suggested from the diffuseness and broadness of the absorption spectra in the dA.dT base pair containing polymers. The preference of formation of site II adducts in dG-rich sequences in the case of the biologically highly active (+)-anti-BPDE isomer is discussed in terms of the known binding and mutation spectra.

[An epidemiological study on syphilis conducted at the laboratory of the Mauriziano Hospital (USL 37) of Lanzo Torinese in 1988]. / Indagine epidemiologica sulla sifilide condotta presso il laboratorio dell'Ospedale Mauriziano di Lanzo Torinese (USL 37) nell'anno 1988.

The study conducted in 1988 was designed to assess the value of blanket serum tests for syphilis on hospitalised and outpatients. The incidence of syphilis encountered was particularly low and primarily confined to patients over 50. Apart from specific requests, mass scanning for the disease does not therefore seem necessary though it does seem advisable to test hospitalised and pregnant women.

[Prevalence of HBsAG carriers in a population of Northern Italy in 1987]. / Prevalenza dei portatori di HBsAG in una popolazione del Nord Italia relativa all'anno 1987.

In view of the high incidence of hepatitis B in Italy, the prevalence of carriers of HBsAg in one area of Piedmont and comparison with data obtained in other areas of Northern and Southern Italy has been assessed. HBsAg was sought in the serum of 2242 patients of the Lanzo Torinese Hospital. Average prevalence observed (0.66%) was related to recent national statistics and shows a slightly lower percentage than mean values for Northern Italy and much below what is found in the South of Italy.

Program evaluation research in ongoing alcoholism treatment: a summary of the Tuscaloosa VA project.

Early in 1980 a Veterans Administration Medical Center initiated a comprehensive program evaluation project of its Alcohol Treatment Unit. The project has resulted in numerous publications detailing the major findings; however, many outcomes of interest have not been reported, and no report is extant which summarizes the outcome of the program evaluation. In this paper, many results are reported for the first time, and previously published results are briefly described. Additionally, the problems encountered in program evaluation are discussed, as are the implications of the results of the project.

[Programming VTIM physiologic pacemakers (evoked QT)]. / Programación de los marcapasos fisiológicos VTIM (QT evocado).

Seventeen VTIM pacers were implanted in seventeen patients. Average follow up were 16.7 months. This paper concerns TX track and TX on modes only. Sensing window period and slope, which determine the rate response, must be programmed carefully. Potential competitive rhythms, severe ventricular arrhythmias or inadequate rate response to physical stress may potentially develop under inadequate programming. Consequently, it is advisable to program the sensing window 20-25 ms longer than the QTE max (at LRL) and the slope at 100% of the whole value. No complications were detected. Figures 1, 2 and 3 show examples of competitive rhythms without heart stimulation and unsensed beats by abnormally long programmed SW values. When properly shortened, this interval allows proper measurement of the Stim.-T and no competitive rhythm, during almost the same spontaneous rhythm. This system fulfills the major conditions for pacing the heart: rate adaptation to neurohormonal release, to physical stress and operative mode as a close-loop system during the recovery period.