Prevention of healthcare associated infections: a descriptive study.

BACKGROUND: This study aims to verify whether there are, and to which degree, knowledge and adherence to guidelines on the prevention and control of healthcare associated infections by nursing staff. Study design. A descriptive study was conducted on a sample of nurses in the areas of medicine, surgery, and its own specialties of the Istituto di Ricovero e Cura a Carattere Scientifico Ca' Granda Ospedale Maggiore Policlinico in Milan from 1st December 2015 to 29th February 2016. METHODS: The knowledge of the nursing staff have been investigated through the use of questionnaires with anonymous self-reporting method; inspections in the wards using observational grids were carried out in order to verify adherence to best-practice principles. The data collected concern, both for the knowledge and for the practice, the following macro-areas: a) Cleaning, disinfection and sterilization, b) Hand hygiene, c) Standard and isolation precautions, d) Prevention of catheter-related urinary tract infections, e) Prevention of catheter-related bacteremia, f) Prevention of surgical site infections, g) Prevention of respiratory tract infections. Statistical analyzes were performed using Microsoft Office Excel and STATA software. RESULTS: 245 nurses from 16 wards were involved. In each wards 4 inspections were conducted. 128 completed questionnaires were returned, all considered for the analysis of data; the adhesion was 52.2%. The participants achieved an overall score of 15.0 ± 4.1 (mean ± SD) on a maximum achievable score of 23 and >75% of them have reached a sufficient level. Among the most positive results, it must be underlined that nurses have demonstrated a higher level of knowledge for hand hygiene, with >81% correct answers; that the lumens of central venous catheters, when not in use, were kept covered with a protective cap in more than 99% of cases; that, for patients bearers of urinary catheter, the urinary drainage bag was maintained below the level of the bladder, as recommended, in more than 91% of the cases. On the contrary, as a very negative result, we found the greatest knowledge gap as regards cleaning, disinfection and sterilization, with a number of incorrect answers approaching 50%; furthermore, 64% of nurses wore jewels on their wrists, and / or hands when in action; finally, the alcohol-based handrub device could be easily reached from at least one of the beds of the room in less than 13% of the cases. CONCLUSION: Some knowledge gaps and differences with respect to adherence to best-practice principles for the prevention and control of healthcare associated infections was highlight by the present study.

Reirradiation of salivary gland tumors with carbon ion radiotherapy at CNAO.

AIMS: To report oncologic and functional outcomes in terms of tumor control and toxicity of carbon ion radiotherapy (CIRT) in reirradiation setting for recurrent salivary gland tumors at CNAO. METHODS: From November 2013 to September 2016, 51 consecutive patients with inoperable recurrent salivary gland tumors were retreated with CIRT in the frame of the phase II protocol CNAO S14/2012C for recurrent head and neck tumors. RESULTS: Majority of pts (74.5%) had adenoid cystic carcinoma, mainly rcT4a (51%) and rcT4b (37%). Median dose of prior photon based radiotherapy was 60 Gy. Median dose of CIRT was 60 Gy [RBE] at a mean of 3 Gy [RBE] per fraction. During reirradiation, 19 patients (37.3%) experienced grade G1 toxicity, 19 pts (37.3%) had G2 and 2 pts (3.9%) had G3. Median follow up time was 19 months. Twenty one (41.2%) patients had stable disease and 30 (58.8%) tumor progression at the time of last follow up. Furthermore, 9 (18%) patients had G1 late toxicity, 19 (37%) had G2 and 9 (17. 5%) had G3. Using the Kaplan Meier method, progression free survival (actuarial) at one and two years were 71.7% and 52.2% respectively. Estimated overall survival (actuarial) at one and two years were 90.2% and 64%, respectively. CONCLUSIONS: CIRT is a good option for retreatment of inoperable recurrent salivary gland tumors with acceptable rates of acute and late toxicity. Longer follow up time is needed to assess the effectiveness of CIRT in reirradiation setting of salivary gland tumors.

Structure of L-aspartate oxidase: implications for the succinate dehydrogenase/fumarate reductase oxidoreductase family.

BACKGROUND: Given the vital role of NAD+ in cell metabolism, the enzymes involved in bacterial de novo NAD+ biosynthesis are possible targets for drug design against pathogenic bacteria. The first reaction in the pathway is catalysed by L-aspartate oxidase (LASPO), a flavoenzyme that converts aspartate to iminoaspartate using either molecular oxygen or fumarate as electron acceptors. LASPO has considerable sequence homology with the flavoprotein subunits of succinate dehydrogenase (SDH) and fumarate reductase (FRD). RESULTS: The crystal structure of the apoform of LASPO from Escherichia coli has been determined to 2.2 A resolution. The enzyme shows a novel fold for an FAD-dependent protein, comprising a three-domain structure: an FAD-binding domain with the dinucleotide-binding fold, a C-terminal three-helical bundle domain, and an alpha + beta capping domain, which is topologically similar to the small subunit of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase. The interface between the FAD-binding and capping domains defines a cleft in which the active site is located. CONCLUSIONS: A number of strictly conserved residues present in all three domains indicate that LASPO, SDH and FRD share the same overall folding topology. Many of these conserved residues are in the FAD-binding site and active centre, suggesting a similar catalytic mechanism. Thus, LASPO, SDH and FRD form a class of functionally and structurally related oxidoreductases that are all able to reduce fumarate and to oxidise a dicarboxylate substrate.

Amino acid sequence around the pyridoxal 5'-phosphate binding sites of 6-phosphogluconate dehydrogenase.

6-Phosphogluconate dehydrogenase (6-phospho-D-gluconate: NADP+ 2-oxidoreductase(decarboxylating), EC 1.1.1.44) from Candida utilis is inhibited by reaction with pyridoxal 5'-phosphate. The aldehydic group of this compound forms a Schiff base with the epsilon-amino group of a lysine residue: reduction of this enamine with tritiated borohydride can label this amino acid. Two tryptic peptides, TS2 and TS3, have been isolated from the labelled protein and found to have the following amino acid sequences: TS2: Ile-Leu-Asx-Glx-Ala-Gly-Gly-Lys(P-Pxy)-Gly-Glx-Thr-Lys TS3: Thr-Val-Ser-Lys(P-Pxy)-Val-Asp-His-Phe-Ile-(Glx,Asx,Glx)-Ala-Lys where Lys(P-Pxy) indicates the modified lysine residue. The similarities between the amino acid sequences around the pyridoxal phosphate binding lysines of 38 peptides, obtained from enzymes which have pyridoxal phosphate as cofactor or inhibitor, are discussed and a prediction is made on the presence of reverse turns in these peptides.

Purification of beef kidney D-aspartate oxidase overexpressed in Escherichia coli and characterization of its redox potentials and oxidative activity towards agonists and antagonists of excitatory amino acid receptors.

The flavoenzyme d-aspartate oxidase from beef kidney (DASPO, EC 1.4. 3.1) has been overexpressed in Escherichia coli. A purification procedure, faster than the one used for the enzyme from the natural source (bDASPO), has been set up yielding about 2 mg of pure recombinant protein (rDASPO) per each gram of wet E. coli paste. rDASPO has been shown to possess the same general biochemical properties of bDASPO, except that the former contains only FAD, while the latter is a mixture of two forms, one active containing FAD and one inactive containing 6-OH-FAD (9-20% depending on the preparation). This results in a slightly higher specific activity (about 15%) for rDASPO compared to bDASPO and in facilitated procedures for apoprotein preparation and reconstitution. Redox potentials of -97 mV and -157 mV were determined for free and l-(+)-tartrate complexed DASPO, respectively, in 0.1 M KPi, pH 7.0, 25 degrees C. The large positive shift in the redox potential of the coenzyme compared to free FAD (-207 mV) is in agreement with similar results obtained with other flavooxidases. rDASPO has been used to assess a possible oxidative activity of the enzyme towards a number of compounds used as agonists or antagonists of neurotransmitters, including d-aspartatic acid, d-glutamic acid, N-methyl-d-aspartic acid, d,l-cysteic acid, d-homocysteic acid, d, l-2-amino-3-phosphonopropanoic acid, d-alpha-aminoadipic acid, d-aspartic acid-beta-hydroxamate, glycyl-d-aspartic acid and cis-2, 3-piperidine dicarboxylic acid. Kinetic parameters for each substrate in 50 mM KPi, pH 7.4, 25 degrees C are reported.

Structural studies on the subunits of glutamate synthase from Azospirillum brasilense.

The amino acid composition and the N-terminal sequences of the two dissimilar subunits of glutamate synthase from Azospirillum brasilense have been determined along with the sequences of selected CNBr peptides. Comparison of our data with those available for Escherichia coli glutamate synthase revealed an overall good homology between the enzymes from the two sources. This is more evident for the heavy subunits where the highly conserved N-terminal sequence containing Cys-1, suggests that this region may be involved in catalysis. However, it appears that the light subunits are different with respect to both their amino acid composition and their N-terminal region, suggesting that the latter may not be part of the enzyme active site. Finally, an extinction coefficient at 444 nm of 62.66 +/- 4.61 mM-1.cm-1 was determined.

cDNA cloning and Escherichia coli expression of UK114 tumor antigen.

Experimental evidence indicates that the antineoplastic effects of UK101, a goat liver perchloric acid extract, is likely due to one of its constituent proteins: the 14 kDa protein named UK114. The cDNA encoding UK114, obtained by PCR methodologies, contains an open reading frame coding for a protein of 137 amino acids with a theoretical molecular mass of 14298 Da. It shows high sequence homology with a 14 kDa protein identified in human, rat and Mus musculus tissues which is likely involved in the inhibition of cell-free protein synthesis. Northern blot analysis indicated that the transcript is present in variable amounts in a wide range of human tissues. Genomic Southern blots revealed that the UK114 mRNA in goat as well as in human is encoded by a single gene, as is the case in rat. The expression system for UK114 was constructed under the control of the PL promoter from bacteriophage lambda and the cDNA coding region has been highly expressed in Escherichia coli as a thioredoxin fusion protein. The recombinant UK114, purified to homogeneity, is immunoreactive to rabbit antisera prepared against UK101 or native UK114, as well as to sera of UK101-treated cancer patients. It inhibits cell-free protein synthesis at 8 microM concentration.

Structural and functional characterization of subunits of the F0 sector of the mitochondrial F0F1-ATP synthase.

Proteolytic digestion of F1-depleted submitochondrial particles (USMP), reconstitution with isolated subunits and titration with inhibitors show that the nuclear-encoded PVP protein, previously identified as an intrinsic component of bovine heart F0 (F01) (Zanotti, F. et al. (1988) FEBS Lett. 237, 9-14), is critically involved in maintaining the proper H+ translocating configuration of this sector and its correct binding to the F1 catalytic moiety. Trypsin digestion of USMP, under conditions leading to cleavage of the carboxyl region of the PVP protein and partial inhibition of transmembrane H+ translocation, results in general loss of sensitivity of this process to F0 inhibitors. This is restored by addition of the isolated PVP protein. Trypsin digestion of USMP causes also loss of oligomycin sensitivity of the catalytic activity of membrane reconstituted soluble F1, which can be restored by the combined addition of PVP and OSCP, or PVP and F6. Amino acid sequence analysis shows that, in USMP, modification by [14C] N,N'-dicyclohexylcarbodiimide of subunit c of F0 induces the formation of a dimer of this protein, which retains the 14C-labelled group. Chemical modification of cysteine-64 of subunit c results in inhibition of H+ conduction by F0. The results indicate that proton conduction in mitochondrial F0 depends on interaction of subunit c with the PVP protein.

Isolation, characterization and partial sequence of cyanogen bromide fragments and thiol peptides from pig kidney D-amino-acid oxidase.

A partial characterization of the primary structure of D-amino-acid oxidase (D-Amino-acid:oxygen oxidoreductase (deaminating), EC 1.4.3.3.) from hog kidney has been achieved by a CNBr cleavage of the 14C-carboxymethylated protein. Four fragments have been isolated and purified and their alignment made possible by overlapping with methionine-containing peptides derived from tryptic digestion of the 14C-carboxymethylated protein. A partial sequencing of the CNBr fragments has been carried out by the automated Edman procedure and by manual sequence analysis. Chymotryptic peptides containing the 5 alkylated thiols of the monomer enzyme (Curti, B., Ronchi, S., branzoli, U., Ferri, G. and Williams, Jr., C. H. (1973) Biochim. Biophys. Acta 327, 266-273) have been isolated and their sequence determined. The present results do not show any significant homologies with the known sequences of other flavoproteins.

The intron-containing L3 ribosomal protein gene (RPL3): sequence analysis and identification of U43 and of two novel intronic small nucleolar RNAs.

Isolation and sequencing of bovine and human intron-containing L3 ribosomal protein genes are here reported. They exhibit very similar organisation, both comprising 10 exons and nine introns. A polymorphic locus, involving a 19-bp deletion, was found in intron 6 of the human gene. The frequency of the two alleles has been estimated in 200 haploid genomes. In bovine and human genes intron sequences are rather different, except for limited regions, located in corresponding positions, which show a surprisingly high degree of identity. All these regions contain conserved features defining the box C/D class of small nucleolar RNAs. Demonstration is given that U43 small nucleolar RNA is encoded within the first intron of both bovine and human genes. Single nucleotide sequences, encoding two novel species of small nucleolar RNAs (U82, U83a and U83b), are located in introns 3, 5 and 7. Their expression has been investigated and a possible role of these molecules in 2'-O-ribose methylation of rRNAs is discussed.

cDNA sequence for bovine ribosomal protein L3 carrying a bipartite nuclear targeting motif, identified also in many other ribosomal proteins.

A full-length cDNA encoding bovine ribosomal protein L3 was isolated and sequenced. The deduced protein sequence comprises 403 amino acids and shows a high level of identity with the other known mammalian L3 proteins. Southern blot analysis of bovine genomic DNA suggests that the bovine genome contains at least 4 copies of the L3 gene. A single hybridisation band of about 1.3 kb is detectable by Northern blot analysis. Within the amino acid sequence, two potential nuclear targeting sequences were detected: one at the N-terminal end and the other, consisting in a bipartite motif (amino acids 341 to 358), present and not previously noticed also in the other known mammalian L3 proteins. A search on all the available mammalian ribosomal proteins revealed the presence of this bipartite motif in many of these proteins.

Aldose reductase is involved in long-term adaptation of EUE cells to hyperosmotic stress.

Aldose reductase has been shown to be expressed in large amount by human embryonic epithelial cells (EUE) in response to osmotic stress. This conclusion is the result of studies undertaken following the purification to homogeneity of two forms of a 35-kDa protein overexpressed in EUE cells grown in hypertonic saline culture medium as compared to EUE cells grown in isoosmotic medium. Amino-acid composition, molecular weight and partial internal amino-acid sequence showed that the above proteins are two different forms of aldose reductase. These findings were confirmed by the observation that aldose reductase activity increased about 150-fold in adapted cells and returned to basal levels in de-adapted cells.

Properties of the flavoenzyme D-aspartate oxidase from Octopus vulgaris.

The properties of D-aspartate oxidase from Octopus vulgaris (EC 1.4.3.1) have been investigated. The protein is a monomer of M(r) 37,000 containing one mol flavin/mol protein. The enzyme as isolated exists at least in two forms, one containing FAD and the other, which is catalytically inactive, probably containing 6-OH-FAD, as inferred from the absorption spectrum of the enzyme. An additional form of the enzyme, as far as the nature of the coenzyme is concerned, has been detected in the purified enzyme and shown to derive from the form originally containing FAD. The modulation of the coenzyme reactivity exerted by Octopus D-aspartate oxidase, as studied by spectrophotometric techniques, conforms to the one expected for an enzyme belonging to the oxidase class of flavoproteins. Structural investigations show similarities in both the amino-acid composition and the N-terminal amino-acid sequence to bovine D-aspartate oxidase and porcine D-amino-acid oxidase. In summary, the general properties of the enzyme from Octopus vulgaris closely resemble those of the enzyme from beef kidney. Moreover, kinetic analyses suggest that two active-site residues with pKa of 7.1 and 9.1 are critical for catalysis, and that the ionization of such residues has different effects on the catalytic activity depending whether mono- or dicarboxylic D-amino acids are used as substrate.

[Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis]. / Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística.

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7%), P. aeruginosa (37.4%) and Haemophilus spp (15.3%). A high resistance to erithromycine (35.0%) and clindamicine (29.4%) was observed in S. aureus strains and 25.9% of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0%). The rate of ampicillin-resistant Haemophilus spp. was 23.0% and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0%).

Infusão de morfina e cetamina, associada ou não à lidocaína, em gatas submetidas à ovariossalpingo-histerectomia / Infusion of morphine and ketamine, with or without lidocaine, in cats undergoing ovariohysterectomy

O objetivo deste estudo foi avaliar os efeitos analgésicos transoperatórios da infusão contínua de morfina e cetamina, associada ou não à lidocaína, em gatas submetidas à OSH eletiva. Foram utilizadas 16 fêmeas adultas, hígidas, pré-medicadas com acepromazina (0,1mg/kg) e morfina (0,5mg/kg), ambas pela via intramuscular, induzidas com cetamina (1mg/kg) e propofol (4mg/kg), pela via intravenosa, e mantidas sob anestesia geral inalatória com isoflurano a 1,4 V%. Os animais foram alocados aleatoriamente em dois grupos: grupo morfina, lidocaína e cetamina (MLK, n=8), que recebeu bolus de lidocaína (1mg/kg), pela via IV, seguido de infusão de morfina, lidocaína e cetamina (0,26mg/kg/h, 3mg/kg/h e 0,6mg/kg/h, respectivamente); e grupo morfina e cetamina (MK, n=8), que recebeu bolus de solução salina, seguido de infusão de morfina e cetamina, nas mesmas doses do MLK. Os momentos avaliados foram: M0, basal, cinco minutos após a indução; M1, imediatamente após a aplicação do bolus de lidocaína ou solução salina; M2, M3, M4 e M5, a cada cinco minutos, até completar 20 minutos do início da infusão; M6, após a incisão da musculatura; M7, após pinçamento do primeiro pedículo ovariano; M8, após pinçamento do segundo pedículo ovariano; M9, após pinçamento da cérvix; M10, após sutura da musculatura; M11, ao final da cirurgia; e M12, M13 e M14, intervalos de cinco minutos, até completar uma hora de infusão. A FP no M0 foi maior no MLK quando comparado ao MK. Em ambos os grupos, a PAS foi maior no M7 e no M8 em relação ao M0, porém no MK, além da PAS, a FP foi maior do M7 ao M13, assim como a f. Os animais do MK necessitaram de um número maior de resgates transoperatorios, total de 23, do que o MLK, total de sete. Conclui-se que a adição de lidocaína incrementou a analgesia oferecida, reduzindo o número de resgates analgésicos transoperatórios, a dose total de fentanil, bem como a probabilidade de os animais necessitarem dese tipo de resgate.(AU)

The aim of this study was to evaluate the trans-operative analgesics, continuous infusion of morphine and ketamine, with or without lidocaine in cats undergoing elective OSH. Sixteen adult cats were used, otherwise healthy, pre-medicated with acepromazine (0.1mg/kg) and morphine (0.5mg/kg), both intramuscularly, induced with ketamine (1mg/kg) and propofol (4mg/kg), intravenous, maintained under general inhalation anesthesia with isoflurane 1.4 V%. The animals were randomly allocated into two groups: morphine, lidocaine and ketamine (MLK, n= 8), which received intravenous bolus of lidocaine (1mg/kg) followed by infusion of morphine, lidocaine and ketamine (0.26mg / kg/h, 3mg / kg/h and 0.6mg / kg/h, respectively); Morphine and ketamine (MK, n= 8), who received bolus of saline followed by infusion of morphine and ketamine at the same doses of MLK. The evaluated moments were: M0, basal, 5 minutes after induction; M1 immediately after the application of lidocaine bolus injection or saline; M2, M3, M4 and M5, every 5 minutes to complete 20 minutes after the start of infusion; M6, after the incision of the musculature; M7, after clamping of the first ovarian pedicle; M8, after clamping of the second ovarian pedicle; M9, after clamping of the cervix; M10, after suturing of the musculature; M11, at the end of surgery; And M12, M13 and M14, 5 minute intervals until completing one hour of infusion. The time to extubating and full recovery of animals, and the need for rescue analgesic fentanyl intraoperatively were also evaluated. HR in M0 was higher in MLK when compared to MK. In both groups the SBP was higher in M7 and M8 compared to M0, but the MK, addition of SAP, HR was greater M7 to M13, as well as f. MK animals required a greater number of trans-operative rescues than the MLK. It was concluded that the addition of lidocaine to the protocol using morphine and ketamine increased its analgesia.(AU)

Efeito de frações inspiradas de oxigênio e modalidades ventilatórias diferentes sobre a idade de cães / [Effects of oxygeninspired fractions and different ventilatory modalities on the age of dogs]

Objetivou-se avaliar diferentes modalidades ventilatórias em cães de diferentes idades submetidos à fração inspirada de oxigênio (FiO2) de 40% e 100%. Foram utilizados 36 cães de três grupos etários (GJ: 0-5; GA: 5-10 e GG: 10-15 anos), sem padronização de peso, sexo, raça e procedimento cirúrgico. Foram pré-medicados com acepromazina e morfina (0,02 e 0,5mg/kg), induzidos à anestesia geral com propofol dose-efeito, manutenção do plano anestésico com isoflurano em 1,3 V% e fornecimento de oxigênio conforme a FiO2 estabelecida para o grupo. Os animais foram submetidos a quatro diferentes modalidades ventilatórias: ventilação espontânea (VE), ventilação ciclada a volume (VCV), ventilação ciclada a pressão (VCP) e ventilação ciclada a pressão com PEEP (VCPP), e permaneceram 30 minutos em cada modalidade. Os parâmetros cardiovasculares mantiveram-se estáveis para todas as FiO2, modalidades ventilatórias e idades. Com relação aos parâmetros ventilatórios, na FiO2 100%, foram observados PaCO2 de 45mmHge e 29% de shunt, enquanto a FiO2 40% apresentou PaCO2 de 43 mmHg e 13% de shunt. Em relação às diferentes idades, os animais adultos e geriátricos apresentaram maiores valores de shunt (26% e 22%) e PaCO2 (44mm/Hg e 46mm/Hg). Conclui-se que a fração inspirada de 40% e a modalidade ventilatória ciclada a volume mostraram-se mais eficientes.(AU)

The objective was to evaluate different ventilatory modalities in dogs of different ages submitted to the inspired fraction of oxygen (FiO2) of 40% and 100%. Thirty-six dogs from three age groups (GJ 0-5, GA 5-10 and GG 10-15 years) were used, without standardization of weight, gender, race and surgical procedure. They were premedicated with acepromazine and morphine (0.02 and 0.5mg/kg), induced to general anesthesia with propofol dose/effect, maintenance of the anesthetic plane with isoflurane in 1.3V% and oxygen supply according to FiO2 established for the group. The animals were submitted to 4 different ventilation modalities, spontaneous ventilation (VS), volume-cycled ventilation (VCV), pressure-cycled ventilation (VCP) and pressure-cycled ventilation with PEEP (VCPP) and remained 30 minutes in each modality. The cardiovascular parameters remained stable for all FiO2, ventilatory modalities and ages. Regarding ventilatory parameters, in FiO2 and PaCO2 of 45mmHg and 29% of shunt, in FiO2 100%, PaCO2 of 43mmHg and 13% of shunt were observed. Regarding the different ages, adult and geriatric animals presented higher values of Shunt (26 and 22%) and PaCO2 (44 and 46mmHg). It was concluded that the inspired fraction of 40% and the volume-cycled ventilatory modality were more efficient.(AU)

Salvage image-guided intensity modulated or stereotactic body reirradiation of local recurrence of prostate cancer.

OBJECTIVE: To retrospectively evaluate external beam reirradiation (re-EBRT) delivered to the prostate/prostatic bed for local recurrence, after radical or adjuvant/salvage radiotherapy (RT). METHODS: 32 patients received re-EBRT between February 2008 and October 2013. All patients had clinical/radiological local relapse in the prostate or prostatic bed and no distant metastasis. re-EBRT was delivered with selective RT technologies [stereotactic RT including CyberKnife(TM) (Accuray, Sunnyvale, CA); image-guidance and intensity-modulated RT etc.]. Toxicity was evaluated using the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer criteria. Biochemical control was assessed according to the Phoenix definition (NADIR + 2 ng ml(-1)). RESULTS: Acute urinary toxicity: G0, 24 patients; G1, 6 patients; G2, 2 patients. Acute rectal toxicity: G0, 28 patients; G1, 2 patients; and G2, 1 patient. Late urinary toxicity (evaluated in 30 cases): G0, 23 patients; G1, 6 patients; G2, 1 patient. Late renal toxicity: G0, 25 patients; G1, 5 patients. A mean follow-up of 21.3 months after re-EBRT showed that 13 patients were free of cancer, 3 were alive with biochemical relapse and 12 patients were alive with clinically evident disease. Four patients had died: two of disease progression and two of other causes. CONCLUSION: re-EBRT using modern technology is a feasible approach for local prostate cancer recurrence offering 2-year tumour control in about half of the patients. Toxicity of re-EBRT is low. Future studies are needed to identify the patients who would benefit most from this treatment. ADVANCES IN KNOWLEDGE: Our series, based on experience in one hospital alone, shows that re-EBRT for local relapse of prostate cancer is feasible and offers a 2-year cure in about half of the patients.

Isolation of the epithiospecifier protein from oil-rape (Brassica napus ssp. oleifera) seed and its characterization.

The epithiospecifier protein (ESP) is a myrosinase (MYR) cofactor, which is necessary to drive the MYR-catalyzed hydrolysis of some specific glucosinolates towards the production of cyanoepithioalkanes instead of isothiocyanates and nitriles. ESP was isolated from Brassica napus seeds by anionic exchange and gel filtration chromatography. ESP showed a molecular weight of about 39 kDa and pI 5.3. The amino acid sequence of several tryptic peptides of ESP (accounting for about 50% of the total sequence) made it possible to establish the high similarity (81% identity) with a hypothetical 37 kDa protein (TrEMBL data base accession number Q39104) and several jasmonate-inducible proteins from Arabidopsis thaliana. This observation suggests that ESP is likely to be involved in jasmonate-mediated defence and disease resistance mechanisms.

Molecular cloning of the cDNA coding for Xenopus laevis prion protein.

Isolation and characterization of the cDNA coding for the 216-residue Xenopus laevis prion protein is reported. Existence of this protein in amphibians was suggested by an EST fragment (accession number BG813008), while a conclusive demonstration is presented here. This protein exhibits a higher identity level to avian and turtle prion (more than 44%) than to mammalian prion (about 28%). Although most of the structural motifs common to known prion proteins are conserved in X. laevis, the lack of repeats represents a substantial difference. Other features worth noting are the presence of not perfectly conserved hydrophobic stretch, which is considered the prion signature, as well as the complete absence of histidine residues.