An attractive way of egg white protein by-product use for producing of novel anti-hypertensive peptides.

The aim of this study was to (i) examine how enzymatic hydrolysis with a non-commercially available proteinase of fig-leaf gourd fruit (Cucurbita ficifolia) increased the use value of egg white protein preparations, generated as byproducts in the industrial process of lysozyme and cystatin isolation from egg white, and (ii) evaluate the inhibition of angiotensin I-converting enzyme (ACE) by the obtained hydrolysates. Purification procedures including membrane filtration, gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC) led to the production of several peptide fractions. Two novel ovalbumin-derived tetrapeptides: SWVE (f 148-151) and DILN (f 86-89) with ACE inhibitory activity were obtained. Study of their inhibitory kinetics revealed a non-competitive binding mode, with an IC50 value against ACE of 33.88 and 73.44 µg for SWVE and DILN, respectively. Synthetic peptides which were designed on the basis of peptide SWVE were examined. A tripeptide sequence of SWV revealed the strongest ACE-inhibitory activity.

Effect of dietary fish oil on milk yield, fatty acids content and serum metabolic profile in dairy cows.

The aim of the study was to evaluate the effects of dietary unprotected fish oil on milk yield, fatty acids content and serum metabolic profile in dairy cows. Forty lactating Polish Holstein-Friesian cows were assigned to two groups. The cows were fed a control basal diet (C group), or a basal diet containing 2% addition of fish oil on mineral carrier (FOM group) during a 8-week period. Supplementing FOM diet of dairy cows had no significant effect on milk fat, milk protein, milk lactose concentration and somatic cells count, but increased the milk yield (36.5 kg/d), (p < 0.05) compared with the control cows (34.28 kg/d). We observed an increase (p < 0.05) in body condition scoring between C and FOM groups. Additionally, higher levels of non-esterified fatty acids and ß-hydroxybutyrate acid were noticed in FOM group after 8 weeks of the experiment; however, these serum parameters still were in the reference range. Concentration of insulin was higher (30.40 µU/ml; p < 0.01) in the FOM group compared with the control group (14.03 µU/ml). In the FOM group, significant increase (p < 0.01) in long-chain fatty acids, mainly cis-9, trans-11 CLA and n-3 fatty acids (eicosapentaenoic and docosahexaenoic acids) was observed. The obtained results demonstrated that addition of fish oil to cows' diet could profitably modify the fatty acids in cow's milk.

The perioperative specialist practitioner: developing and evaluating a new surgical role.

INTRODUCTION: This paper describes the design, implementation and evaluation of a new professional role in surgery. The role of the perioperative specialist practitioner (PSP), conceived as a response to the Working Time Directive, provides integrated preoperative and postoperative care to patients undergoing surgery in hospital. METHODS: A 1-year training programme was designed, dealing with a wide range of knowledge, skills and attitudes. Effective communication was a key component. Nine intensive 5-day modules at Imperial College London (London, UK) alternated with supervised experience of the surgical team at each participant's home trust. Detailed evaluation of the role and the training programme was provided by an independent research team, using an interview-based qualitative approach. Observational data were provided by the project team. Data were analysed using standard qualitative methods. RESULTS: 27 PSPs across 12 National Health Service trusts took part in two PSP training programmes. A total of 124 interviews (94 individual and 30 group) were carried out with PSPs and their colleagues. Overall, the role was seen as successful and positive, with great potential for dealing with reductions in junior medical cover. Each site encountered different opportunities and problems. Lack of mentorship was a key issue, and the role provoked considerable opposition in trusts. The training programme was viewed as highly successful. DISCUSSION: PSPs can provide high levels of expertise, but within clear limits. Our training programme has been effective and is perceived to be of high quality. However, introducing a new role requires time and sensitivity if opposition is to be minimised.

Innovative training for new surgical roles--the place of evaluation.

BACKGROUND: This paper describes the central role of 'external' evaluation, provided by an independent researcher, in developing an innovative curriculum for new professional roles with surgery. Workforce changes affecting the National Health Service provided an opportunity to develop 2 new roles and design training programmes to support them. The perioperative specialist practitioner (PSP) role was designed from scratch, while surgical care practitioner (SCP) training built on existing practice. Training programmes combined formal modules at Imperial College London (approximately 48 days over 10 months) with supervised clinical practice in each participant's base hospital. Programmes balanced factual knowledge, clinical and communication skills, professional issues and personal development and used a range of innovative techniques. EVALUATION METHODS: A qualitative approach based on a utilisation-focused model monitored the development and implementation of 4 pilot PSP and SCP training programmes. A total of 124 individual and 48 group interviews were conducted at intervals over 3 years, sampling course participants, the project team clinical supervisors and administrators. An independent researcher collected, analysed and presented data at key stages, feeding back findings to the project team as the programmes evolved. DISCUSSION: Effective training programmes for new roles can be developed, but the process is time-consuming and requires sensitivity. An independent evaluator offers great benefits, modulating the collaborative partnership between participants and project team. Positive responses (relating to content and teaching methods) from our study enabled us to refine a learner-centred programme. Negative responses often demanded immediate action to address important concerns, and evaluation provided early warning. External evaluation provides a vital perspective in the development of curricula supporting new roles.

The profiles of enterotoxin genes in Staphylococcus aureus from nasal carriers.

AIMS: To evaluate the occurrence of enterotoxin genes in Staphylococcus aureus recovered from nasal carriers. METHODS AND RESULTS: Eighty S. aureus strains were tested for the presence of 17 new enterotoxin genes using multiplex-PCR. Sixty-one isolates were found to carry enterotoxin genes. The majority of the enterotoxigenic isolates carried enterotoxin gene cluster (egc) genes, namely seg, sei, sem, sen and seo. The egc type containing the seu gene was found in 19 of the 47 isolates with egc-like genes. Interestingly, no seu-containing egc coexisted with sec and sel, as was the case for a considerable portion of the isolates carrying a seu-negative egc. The tst gene was detected in two isolates carrying sec and sel only and in eight isolates carrying seu, but not in the isolates containing the seu-negative egc type. CONCLUSIONS: The genes forming an egc were found to be predominant in S. aureus from nasal carriers. The coexistence of a seu-positive egc with tst in contrast to an egc lacking the seu gene apparently is not associated with the presence of tst and can reflect a difference between these gene groupings. SIGNIFICANCE AND IMPACT OF THE STUDY: The egc types carried by the analysed isolates seem to have an influence on the distribution of other genes located on staphylococcal pathogenicity islands, which may modulate the repertoire of virulence factors carried by a single S. aureus strain.

Superantigen types in Staphylococcus aureus isolated from patients with cystic fibrosis.

The screening of 17 SAg genes of S. aureus isolated from the sputum of cystic fibrosis (CF) patients revealed that among 47 genetically different strains, 39 (83 %) carried SAg genes. Superantigens forming enterotoxin gene cluster were detected in 20 strains. The 2nd most common superantigen type was selk detected in 13 strains. In 9 strains, selk occurred together with the sea gene. Out of 74 strains recovered from nasal carriers, 56 (75 %) were found to carry SAg genes, 38 carried egc genes, while selk was detected in 5 strains. The predominant SAg types in both investigated S. aureus populations were egc and selk/sea, but selk gene frequency was significantly higher in the CF-derived strains.

Factors affecting killer activity of some yeast species occurring in Rokpol cheese.

Fourteen strains of Candida famata and 7 strains of C. sphaerica originating from blue-veined Rokpol cheese were studied for their ability to produce killer toxins against 3 strains of Yarrowia lipolytica selected as potential starter cultures for cheesemaking. All tested strains revealed killer activity in the presence of salt, at 14 degrees C and pH 4.6. Only one Y. lipolytica strain was resistant to C. famata and C. sphaerica killer toxins.

Identification and biochemical characteristics of yeast microflora of Rokpol cheese.

Yeast microflora on the surface and interior of Rokpol cheese was examined for cheeses produced at three dairies located in Lower Silesia, Poland. Yeast populations on the surface of the cheeses ranged from 10(5)-10(9) cfu/g, but were 10-100 times lower for interior samples. The occurrence and proportions of yeast species varied, depending on the dairy plant and cheese sample. The most frequently isolated species were: Candida famata and C. spherica, followed by C. intermedia and Geotrichum sp. Other species such as Saccharomyces kluyveri, C. kefyr and C. lipolytica were found occasionally. Extracellular and intracellular proteolytic and lipolytic activities were examined for 39 isolates of C. famata.

Aspartic proteinase from Penicillium camemberti: purification, properties, and substrate specificity.

An acid proteinase from the culture filtrate of Penicillium camemberti was isolated in a two-step purification procedure by cation exchange chromatography and gel filtration. The enzyme is an aspartic proteinase inhibited by pepstatin, DAN, and EPNP, with a molecular mass determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 33.5 kDa. The optimum activity for hydrolysis of denatured hemoglobin is around pH 3.4. The enzyme is highly specific for the aromatic and hydrophobic amino acid residue in insulin B-chain and, like pepsin, selectively splits only one Leu7-Met8 peptide bond in the squash trypsin inhibitor CMTI 1. The hydrolyzed bond can be resynthesized by P. camemberti proteinase at neutral pH.

Purification and characterization of a cell wall peptidase from Lactococcus lactis subsp. cremoris IMN-C12.

A peptidase from the cell wall fraction of Lactococcus lactis subsp. cremoris IMN-C12 has been purified to homogeneity by hydrophobic interaction chromatography, two steps of anion-exchange chromatography, and gel filtration. The molecular mass of the purified enzyme was estimated to be 72 kDa by gel filtration and 23 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme has a pI of 4.0, and it has the following N-terminal sequence from the 2nd to the 17th amino acid residues: -Arg-Leu-Arg-Arg-Leu-?-Val-Pro-Gly-Glu-Ileu-Val-Glu-Glu-Leu-Leu. The peptidase is most active at pH 5.8 and at 33 degrees C with trileucine as the substrate. Reducing agents such as dithiothreitol, beta-mercaptoethanol, and cysteine strongly stimulated enzyme activity, while p-chloromercuribenzoate had an inhibitory effect. Also, metal chelators lowered the peptidase activity, which could not be restored with Ca2+ and Mg2+. The divalent cations Cu2+, Zn2+, Fe2+, and Hg2+ completely inhibited peptidase activity. The peptidase is capable of hydrolyzing tripeptides and some dipeptides, with a preference for peptides containing leucine and with the highest activity towards the tripeptides Leu-Leu-Leu, Leu-Trp-Leu, and Ala-Leu-Leu, which were hydrolyzed with Kms of 0.37, 0.18, and 0.61 mM, respectively.