A novel synthetic medium and expression system for subzero growth and recombinant protein production in Pseudoalteromonas haloplanktis TAC125.

The Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a model organism of cold-adapted bacteria. The interest in the study of this psychrophilic bacterium stems from its capability either as a non-conventional system for production of recombinant protein and as a rich source of bioactive compounds. To further explore the biotechnological ability of P. haloplanktis TAC125, we have developed a synthetic medium, containing D-gluconate and L-glutamate (GG), which allows the bacterium to grow even at subzero temperatures. P. haloplanktis TAC125 growing in GG medium at low temperature displays growth kinetic parameters which confirm its spectacular adaptation to cold environment and subzero lifestyle, paving the way to the definition of the underlying molecular strategies. Moreover, in this paper, we report the setup of a finely regulated gene expression system inducible by D-galactose to produce recombinant protein in GG synthetic medium at temperatures as low as -2.5 °C. Thanks to the combination of the novel medium and the new expression system, we obtained for the first time the production of a recombinant protein at subzero temperature, thus providing an innovative strategy for the recombinant production of "difficult" proteins.

DG Connect Funded Projects on Information and Communication Technologies (ICT) for Old Age People: Beyond Silos, CareWell and SmartCare.

Information and communication technologies (ICT) are promising for the long-term care of older and frequently frail people. These innovations can improve health outcomes, quality of life and efficiency of care processes, while supporting independent living. However, they may be disruptive innovations. As all European member states are facing an increasing complexity of health and social care, good practices in ICT should be identified and evaluated. Three projects funded by DG CNECT are related to Active and Healthy Ageing (AHA) and frailty: (i) BeyondSilos, dealing with independent living and integrated services, (ii) CareWell, providing integrated care coordination, patient empowerment and home support and (iii) SmartCare, proposing a common set of standard functional specifications for an ICT platform enabling the delivery of integrated care to older patients. The three projects described in this paper provide a unique pan-European research field to further study implementation efforts and outcomes of new technologies. Below, based on a description of the projects, the authors display four domains that are in their views fundamental for in-depth exploration of heterogeneity in the European context: 1. Definition of easily transferable, high level pathways with solid evidence-base; 2. Change management in implementing ICT enabled integrated care; 3. Evaluation and data collection methodologies based on existing experience with MAST and MEDAL methodologies; and 4. Construction of new models for delivery of health and social care. Understanding complementarity, synergies and differences between the three unique projects can help to identify a more effective roll out of best practices within a varying European context.

Presence and indigenous nature of Lyme disease in southern Italy.

Lyme disease is very common in the countries of the northern hemisphere. In Italy it is endemic in some regions of the northern part of the country and it is more frequent during summer. In Calabria (south Italy) no cases have been reported. To document the presence and indigenous nature of Lyme disease in this territory we conducted a study from 1999 to 2002. We defined as indigenous cases those with erythema migrans with the following characteristics: dimensions equal to or greater than 5 cm; localization on an area of the skin where there was a tick bite; appearance between 4 and 30 days after the tick bite; appearance in patients who had not resided out of Calabria in the previous 3 months. We found 23 patients with the necessary characteristics to be defined indigenous cases. Since 15 of these cases (65.2%) were observed in the October - December trimester and no case was found in the July - September trimester, we suspect that in Calabria the disease follows a seasonal distribution which differs with respect to countries where it is historically endemic.

Antifungal agents. 9. 3-Aryl-4-[alpha-(1H-imidazol-1-yl)arylmethyl]pyrroles: a new class of potent anti-Candida agents.

A new class of potent antifungal agents, namely, 3-aryl-4-[alpha-(1H-imidazol-1-yl)arylmethyl]-pyrroles, is described. These compounds are related to bifonazole and pyrrolnitrin, two compounds belonging to the class of antimycotic drugs. The synthesis of the title pyrroles has been performed starting from 1,3-diaryl-2-propen-1-ones, which were reacted with tosylmethyl isocyanide to give 3-aroyl-4-arylpyrroles. Reduction of the resulting compounds by lithium aluminum hydride furnished the related alcohols, which were treated with 1,1'-carbonyldimidazole to afford the required imidazole derivatives. Forty-four new pyrroles which incorporate an (arylmethyl)imidazole moiety in the 3-arylpyrrole structure were prepared by the above procedure and tested in vitro against Candida albicans and Candida spp. Among test compounds, 10 were found to be highly active against C. albicans. The most active derivative (27) was twice as potent (MIC90) as bifonazole, and its activity was 4 times greater than those of miconazole and ketoconazole. The other nine compounds showed antifungal activity of the same order of that of bifonazole and were ca. 2 times as active as miconazole and ketoconazole. Derivatives 21 and 27 tested in vivo against C. albicans A170 were shown to be highly effective in rabbit skin candidosis. Pharmacological studies on compounds 27 and other related pyrroles (19, 35, 36, 38, 39, and 49) are in progress to select one of them as a potential candidate for clinical experiments.

Antifungal agents. 10. Synthesis and antifungal activities of aryl-(1H-imidazol-1-yl)-(isoquinolin-1-yl) methane derivatives.

Various aryl-(1H-imidazol-1-yl)-(isoquinolin-1-yl)methane derivatives have been synthesized and tested as antifungal agents. The new imidazoles have been obtained by the action of 1,1'-sulfinyldiimidazole on aryl-(isoquinolin-1-yl)carbinols, which have been prepared by standard procedures starting from isoquinoline. Among 44 test derivatives only a few have exhibited some antifungal activity, the most active compound (4e) being twofold less potent than miconazole, ketoconazole and bifonazole, used as standard drugs.

Antifungal agents. VIII. Synthesis and antifungal activities of bipyrryl analogues of bifonazole.

Various bipyrryl analogues of bifonazole were synthesized starting from aryl-3-pyrryl-1-imidazolylmethanes. The introduction of a second pyrryl portion was performed by linking an acrylate moiety at 1-position of the pyrrole ring and then by treatment with TosMIC. The bipyrryl esters were hydrolyzed and decarboxylated to afford the required imidazoles. All new imidazole derivatives were tested against Candida albicans and Candida spp using as standard controls miconazole, bifonazole and ketoconazole.

Antifungal agents. VI. In vitro antifungal activities of halobenzoyl esters of cis- and trans-[2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethyl)-1,3- dioxolan-4-yl]carbinols.

The synthesis and the in vitro antifungal activities against Candida albicans and Candida spp of a number of halobenzoyl esters of cis- and trans- [2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethyl)-1,3-dioxolan -4- yl]carbinols is reported. Some new imidazoles were found more active than ketoconazole and sometimes as potent as bifonazole against Candida albicans. All derivatives were found scarcely active against Candida spp.

Antifungal agents. III. Naphthyl and thienyl derivatives of 1H-imidazol-1-yl-4-phenyl-1H-pyrrol-3-ylmethane.

The in vitro antifungal activities of some naphthyl and thienyl derivatives of 1H-imidazol-1-yl-4-phenyl-1H-pyrrol-3-ylmethane against Candida albicans and Candida spp are reported. The title derivatives were prepared starting from proper arylstirylketones, which were reacted with tosylmethylisocyanide (Tos-MIC) to afford the related 4-phenyl-1H-pyrrol-3-yl aryl ketones. Reduction of ketones to the corresponding carbinols followed by treatment of the last compounds with 1,1'-carbonyldiimidazole (CDI) gave the title imidazoles. The related N-methylpyrrole derivatives are also described.

Antibacterial and antifungal agents. XV. Synthesis and antifungal activity of structural analogues of bifonazole and ketoconazole.

The synthesis and antifungal activities of the cis- and trans-1-acetyl-4-[4-[[2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethy l)- 1,3-dioxolan-4-yl]-methoxy]phenyl)piperazines 3 and 4 are reported. Stereochemical assignments to diastereomeric pairs of cis/trans isomers were made on the basis of 1H- and 13C-NMR data. Among test derivatives the best activity was shown by the benzoyl esters of the cis- and trans-[2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethyl)-1,3-di oxolan-4- yl]methanols 9 and 10.

[Conservative surgery of the sphincters for low rectal cancer]. / Cirugía conservadora de los esfínteres para el cáncer de recto bajo.

A retrospective review was performed of 43 clinical records of patients who underwent surgical treatment for low rectal cancer less than 12 cm from the anus in the General Surgical Service of the Hospital Militar "Dr. Carlos Arvelo" of Caracas, between 1969 and 1988. 21 patients (49%) were males and 22 (51%) females with an average age of 61 years. In 22 (51%) a sphincter saving resection (SSR) was done: 18 low anterior resection, 3 coloanal anastomosis and one pull-through, in 21 (46%) abdominoperineal resection (APER). All were adenocarcinoma except one, reported as a cloacogenic. 2 (9%) operative death were in the SSR and 7 (33%) in the APER. 4 (20%) patients with SSR died without recurrence and 2 (10%) with recurrence. In the group of APER 1 (7%) died without recurrence and 4 (29%) with local recurrence. A critical review was done of our experience and the international literature about the use of the SSR in low rectal cancer.