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The objective of this work was to investigate the structure and diversity of lactic acid bacteria (LAB) communities in sourdough used for the production of traditional breads (Carasau, Moddizzosu, Spianata, Zichi) in Sardinia. 16S rDNA sequencing and Randomly Amplified Polymorphic DNA (RAPD-PCR) was applied for the identification and typing of the LAB isolated from 25 samples of sourdoughs. Multivariate statistical techniques were applied to RAPD-PCR pattern to study the biological diversity of sourdough samples. Twelve different species of LAB were identified, and most isolates were classified as facultative heterofermentative lactobacilli. Lactobacillus pentosus dominated the lactic microflora of many samples while Lactobacillus sanfranciscensis was isolated only from a limited number of samples. Although heterofermentative species represented between between 30% and 60% of the isolates in Carasau, Spianata and Zichi sourdoughs, only 2% of the isolates from Moddizzosu sourdoughs were identified as heterofermentative LAB. RAPD-PCR with a single primer followed by cluster analysis did not allow the identification of the isolates at the species level. However, a multidimensional scaling/bootstrapping approach on the RAPD-PCR patterns uncovered the diversity of the LAB communities of LAB showing differences both within and between bread types.

In order to study the composition of the lactic acid bacteria (LAB) community of sourdoughs used for the manufacture of Altamura bread, a traditional durum wheat bread produced in Apulia (Southern Italy), 111 strains of LAB were isolated and characterized. The phenotypic characterization of the isolates, carried out using a set of 29 tests, allowed the identification of 15 clusters at the 80% similarity level by hierarchical cluster analysis. Of the isolates, 88% were identified as facultatively heterofermentative LAB (Lactobacillus plantarum, Lb. paracasei, Lb. casei) and 12% as heterofermentative LAB (Lb. brevis, Leuconostoc mesenteroides). SDS-PAGE profiles of whole cell proteins of 68 strains confirmed the identification. Both the diversity and structure of the lactic microflora for sourdoughs for Altamura bread varied among samples.

Chemometric analysis of chromatograms plays a fundamental role in characterization of foods or in detection of adulteration. Data for multivariate analysis of chromatographic profiles are usually obtained by visual matching (VM) of peaks, the identities of which, as for peptide profiles from cheese extracts, are often unknown. To avoid the main disadvantages of VM, which is subjective and time-consuming, a novel approach was developed. Fuzzy logic was employed to handle in a systematic way uncertainty in the position of peptide peaks, and chromatograms were processed by a rule-based membership function. Processed data consisted of classes of retention time wherein peak heights were accumulated by using the distance from the center of the class as a weight. The novel approach (fuzzy approach, FA) was compared with VM by using a real data set and by performing multivariate descriptive statistical techniques (principal component analysis, multidimensional scaling, and nonhierarchical cluster analysis). FA provided a fast, reliable, and objective alternative to VM and could be successfully applied for chemometric analysis of chromatographic profiles whenever knowledge of the identity of peaks is lacking or unnecessary.

The dynamic gassing-out method was used to assess the effect of stirrer speed, air flow rate and alginate concentration on the overall volumetric oxygen mass transfer coefficient (K(L)a) in a bench-top stirred fermenter equipped with two Rushton-type turbines. As the alginate concentration in the medium was increased from 0 to 2% (w/v), the liquid film time constant (tau(F)) increased from approx. 30 to 300% of the electrode time constant (tau(E)), clearly showing that the liquid-probe diffusion film affected the oxygen-electrode response. By accounting for delayed probe response, liquid film time constant and gas residence time, the corrected values of K(L)a were empirically correlated with the above independent variables as converted into gassed power input and aeration number per unit liquid volume and the effective liquid/water shear viscosity ratio. Despite an average error of 19%, such a correlation might be useful to adjust the oxygen transfer in bench-top stirred reactors during viscous aerobic fermentations.

Aims: Evaluation of composition and evolution of the coagulase-negative staphylococci (CNS) communities in two traditionally fermented sausages (salsiccia and soppressata lucana) produced in Basilicata, southern Italy., Methods and Results: A culture-dependent approach based on isolation on selective media and identification with phenotypic and molecular methods was used. Phenotypic data of 471 strains were analysed by multivariate statistical methods by using 28 strains from culture collections and 48 strains identified by molecular methods (such as 16S rDNA sequencing, species-specific PCR assays, intergenic spacer region-PCR and PCR-denaturing gradient gel electrophoresis) as a reference. The CNS microflora of the sausages was found to be dominated by different biotypes of Staphylococcus xylosus (51.2%), followed by S. pulvereri/vitulus, S. equorum and S. saprophyticus (13.4, 10.2 and 10%, respectively). Other species (S. succinus, S. pasteuri, S. epidermidis, S. warneri and Macrococcus caseolyticus) were also present at lower levels. Identification of 25% of the isolates was impossible., Conclusions: The composition of CNS communities varied significantly with sausage type, plant and ripening time and clear differences were found among communities of salsiccia and soppressata at the end of ripening., Significance and Impact of the Study: Phenotypic characterization, supported by molecular and statistical analyses, can be considered a useful approach for typing a large number of isolates and for monitoring the evolution of staphylococcal communities during sausage fermentation but does not always provide a satisfactory identification of the isolates.

Introduction: About 40% of patients with Crohns disease (CD) have a perianal involvement. Despite the recent introduction of anti-TNF antibody, this therapy has uncertain long-term results and surgery still remains a major treatment option., Aims & Methods: This study relates our experience in surgical management of perianal CD without anti-TNF treatment. From July 92 to February 02, 37 patients with perianal Crohns disease were treated, 43 underwent local operations or faecal diversion for fistulas and/or abscesses. Patients not requiring surgery or in therapy with anti-TNF. were excluded from the study. We analysed the outcome of surgical treatment for perianal CD., Results: Male to female ratio was 1:0.6, median age was 36,9 years (range 17-62). Perianal disease included 32 fistulas (16 trans-sphincteric, 2 superficial, 2 ano-vaginal, 10 multiple and complex, 2 horseshoe) and 7 abscesses (5 perianal, 2 ischio-rectal). Local surgery included 1 abscess drainage, 5 abscess drainage and fistula incision with seton insertion, 2 fistulotomy, 9 partial fistulectomy and seton insertion. At surgery, 40% of patients were ongoing a medical treatment with 5-ASA and/or antibiotics, 40% with steroids and/or immunosoppressors, 15% only with 5-ASA and 5% no ongoing treatment. The horseshoe fistulas were managed with a fistulotomy and seton insertion. One patient with ano-vaginal fistula required proctectomy and the other one total proctocolectomy. Patients treated by diverting colonostomy (3) had fistula recurrence after its closure in 100%. 20% of patients required total proctocolectomy and ileostomy for extensive intestinal disease. Of the 27 patients undergoing seton insertion or fistulotomy none had faecal incontinence due to the operation and 38% had a 1 year recurrence., Conclusions: Perianal CD is a heterogeneous entity, therefore its management is still controversial. Moreover, a high percentage of patients (18% in our series) requires a major surgery due to the extension and seriousness of rectal involvement. In our survey only 12 patients (39%), with trans-sphincteric fistula, could have been theoretically treated with anti-TNF. We wonder if the cost-and-benefit of this medical treatment justifies its application on patients that could undergo a surgical treatment with good long-term results.

Rapid and reliable identification of Staphylococcus xylosus was achieved by species-specific PCR assays. Two sets of primers, targeting on xylulokinase (xylB) and 60 kDa heat-shock protein (hsp60) genes of S. xylosus, respectively, were designed. Species-specificity of both sets of primers was evaluated by using 27 reference strains of the DSM collection, representing 23 different species of the Staphylococcus genus and 3 species of the Kocuria genus. Moreover, 90 wild strains isolated from different fermented dry sausages were included in the analysis. By using primers xylB-F and xylB-R the expected PCR fragment was obtained only when DNA from S. xylosus was used. By contrast, amplification performed by using primers xylHs-F and xylHs-R produced a single PCR fragment, of the expected length, when DNA from S. xylosus, S. haemolyticus, S. intermedius and S. kloosii were used as template. Nevertheless, AluI digestion of the xylHs-F/xylHs-R PCR fragment allowed a clear differentiation of these 4 species. The rapidity (about 4 h from DNA isolation to results) and reliability of the PCR procedures established suggests that the method may be profitably applied for specific detection and identification of S. xylosus strains.

Introduction: Crohns disease is a panintestinal chronic inflammatory condition. Its remitting-relapsing behaviour may require in the single patient repeated surgeries, with the aim of resolving the complications of the disease. The awereness that surgery cannot resolve the disease has led, in the last years, to the development of new "conservative surgical techniques", which preserve as much of the intestinal tissue as possible. These techniques are minimal resection and strictureplasty (SP). Aim of the study was to perform a prospective analysis of the long-term outcome of SP in a consecutive series of patients undergoing surgery for complicated Crohns Disease at the Division of general surgery, L. Sacco University Hospital, Milano, Italia., Methods and Results: During the period of October 1992 to June 2002, 286 patients underwent surgical procedures for jejunoileal Crohns disease. 116 of them underwent SP resulting in a total of 217 procedures, of which: 111 Heineke-Mikulicz SP (51.2%), 36 ileoileal side-to-side SP (16.6%), 40 ileoceacal SP (18.4%) and the remaining 30 ileocolic SP (13.8%), as previously described by A.M. Taschieri. Fiftyone of the patients (23.5%) had concomitantly a minimal bowel resection. Postoperative mortality was nil, while in 3 cases (2.59%) repeated surgery was necessary due to postsurgical complications. Time-to-event estimates were performed using the Kaplan-Meier function., Conclusions: mortality, morbidity, and long-term results in this population of patients who underwent SP are encouraging and in line with reports in the international literature. It is suggested that SP together with minimal bowel resections, may be considered as first line surgical therapy in patients with Crohns Disease.

Mild lameness is considered a performance-limiting problem that may escape detection until it worsens, and is considered the primary reason for reduced racing performance. The kinematics changes associated with a lame horse at the trot have been demonstrated previously, but the metabolic cost of these alterations in their gait have not been demonstrated. Six fit Thoroughbred horses with an established VO2max participated in 4 trials using a randomised cross-over design study, separated by 10-14 days. The horses were tested with one of 4 trial conditions: lead forelimb lameness (LL); off-lead forelimb lameness (OL); bilateral forelimb lameness (BL) or no lameness (NL). Lameness was induced by sole pressure from a modified shoe that resulted in a consistent slight head nod at a trot in a straight line while jogging in hand. Lameness was adjusted to provide a lameness that would be quantified as a 1-2/5 on the grading system recommended by the AAEP. Each trial consisted of 4 different levels of exercise intensity at speeds equivalent to 30, 60, 80 and 110% of an individual's speed required to elicit VO2max. Stride parameters, oxygen consumption (VO2), carbon dioxide production (VCO2), electrolytes, plasma lactate, glucose and PCV/TP were measured prior to exercise, at each exercise level and after exercise. A multiway ANOVA with repeated measures was utilised to examine possible effects of individual horse, lameness, and exercise intensity on measured parameters. Significance was set at alpha = 0.05. For horses exercising at the maximum intensity, VO2 was significantly lower for both of the single-leg lamenesses (LL or OL) when compared to NL or BL (mean +/- s.e. 165.6 +/- 2.5, 164.7 +/- 3.0, 175.8 +/- 2.4 and 170.9 +/- 2.1 ml O2/min/kg bwt, respectively). Blood lactate concentrations were not significantly different among the treatment groups. However, lactate accumulation rates computed as the change with time in lactate concentration at the highest exercise intensity were significantly higher for LL and OL than for NL and BL (7.8 +/- 03, 83 +/- 0.2, 4.1 +/- 0.2 and 4.7 +/- 0.3 mmol/min, respectively). Exercise intensity had significant effects on all of the measured parameters, but there were no other significant differences due to treatment. These results suggest that metabolic energy transduction is affected by even mild unilateral forelimb lamenesses.

Aims: To evaluate the effect of yeast extract (YE) concentration, temperature and pH on growth and exopolysaccharide (EPS) production in a whey-based medium by Streptococcus thermophilus SY and to characterize the partially purified EPS., Methods and Results: Factorial experiments and empirical model building were used to optimize fermentation conditions and the chemical composition, average molecular weight (MW) and rheological properties of aqueous dispersions of the EPS were determined. Exopolysaccharide production was growth associated and was higher (152 mg l(-1)) at pH 6.4 and 36 degrees C with 4 g l(-1) YE. High performance size exclusion chromatography of the partially purified EPS showed two peaks, with a weight average MW of 2 x 10(6) and 5 x 10(4), respectively. The EPS was a heteropolysaccharide, with a glucose : galactose : rhamnose ratio of 2 : 4.5 : 1. Its water dispersions had a pseudoplastic behaviour and showed a higher viscosity of xanthan solutions., Significance and Impact of the Study: The fermentation conditions and some properties of an EPS produced by Strep. thermophilus, a dairy starter organism, were described.

Objective: To investigate the effects of recurrent laryngeal neurectomy (RLN) in combination with laryngoplasty and ventriculocordectomy on the postoperative performance of Thoroughbred racehorses treated for grade III left laryngeal hemiparesis (LLH)., Study Design: Retrospective study., Sample Population: Fifty-five Thoroughbred racehorses., Methods: Medical records for Thoroughbred racehorses treated surgically by laryngoplasty and laser ventriculocordectomy with or without RLN between June 1993 and December 1996 were reviewed. Outcome was evaluated subjectively by assessment of performance obtained from telephone interviews with owners and trainers, and objectively by assessment of racing performance for 3 races before and after surgery using a performance index (PI)., Results: Fifty-five Thoroughbred racehorses with resting endoscopic grade III LLH were treated by laryngoplasty and ventriculocordectomy either with (39 horses) or without (16 horses) RLN. For RLN horses, respondents for 38 horses believed performance was improved in 19 horses, unchanged in 16 horses, and decreased in 3 horses. For horses without RLN, respondents for 9 horses reported that performance was improved in 5 horses, unchanged in 2 horses, and decreased in 2 horses. PI scores were improved in 18 RLN horses and in 6 horses without RLN; there was no statistical difference in outcome., Conclusions and Clinical Relevance: Use of RLN in combination with laryngoplasty and ventriculocordectomy for treatment of grade III LLH may not improve postoperative racing performance., (Copyright 2001 by The American College of Veterinary Surgeons)

Aims: To evaluate the occurrence and evolution of biogenic amines during ripening of fermented sausages and their relationship with physico-chemical and microbiological properties of the product., Methods and Results: Salsiccia and Soppressata were obtained from artisanal and industrial plants in Basilicata and pH, aW, microbial counts and biogenic amine content were measured. A high variability in amine content was observed. 2-Phenylethylamine and histamine were rarely found, while the tyramine, putrescine and cadaverine content increased during ripening. No correlation was found between individual biogenic amine content, microbial counts or physico-chemical parameters., Conclusion: Starter cultures did not necessarily prevent the production of biogenic amines whose total contents were usually higher in Soppressata, a product with a larger diameter and aW compared with Salsiccia., Significance and Impact of the Study: Literature findings on biogenic amine content and the evolution of microbial populations were confirmed. Normal ranges for amine content in Salsiccia and Soppressata are reported.

Aims: to evaluate the evolution of lactic acid bacteria (LAB) populations in traditional fermented sausages (salsiccia and soppressata) produced in artisanal and industrial plants in Basilicata (Southern Italy)., Methods and Results: Four hundred and fourteen lactic acid bacteria (LAB) cultures were isolated from samples of sausages at different stages of ripening. A phenotypic characterization of the isolates was carried out using a set of 28 tests, and 34 clusters were identified at the 80% similarity level using hierarchical cluster analysis. Of the isolates 50% were identified as Lactobacillus sakei (with several biotypes), 22% as Pediococcus spp. (mainly Ped. pentosaceus), 7% as Leuconostoc (Leuc. carnosum, Leuc. gelidum, Leuc. pseudomesenteroides), 6% as Lact. plantarum, 1% as Lact. curvatus. Other lactobacilli, including unidentified species, were present in lower numbers., Conclusion: The phenotypic diversity and composition of the LAB flora varied as a function of the production plant, product type and ripening time. SIGNIFICANCE AND IMPACT OT THE STUDY: A new procedure based on bootstrapping and Multidimensional Scaling was successfully used to obtain a graphical representation of the evolution of the LAB populations.

Thermophilic streptococci play an important role in the manufacture of many European cheeses, and a rapid and reliable method for their identification is needed. Randomly amplified polymorphic DNA (RAPD) PCR (RAPD-PCR) with two different primers coupled to hierarchical cluster analysis has proven to be a powerful tool for the classification and typing of Streptococcus thermophilus, Enterococcus faecium, and Enterococcus faecalis (G. Moschetti, G. Blaiotta, M. Aponte, P. Catzeddu, F. Villani, P. Deiana, and S. Coppola, J. Appl. Microbiol. 85:25-36, 1998). In order to develop a fast and inexpensive method for the identification of thermophilic streptococci, RAPD-PCR patterns were generated with a single primer (XD9), and the results were analyzed using artificial neural networks (Multilayer Perceptron, Radial Basis Function network, and Bayesian network) and multivariate statistical techniques (cluster analysis, linear discriminant analysis, and classification trees). Cluster analysis allowed the identification of S. thermophilus but not of enterococci. A Bayesian network proved to be more effective than a Multilayer Perceptron or a Radial Basis Function network for the identification of S. thermophilus, E. faecium, and E. faecalis using simplified RAPD-PCR patterns (obtained by summing the bands in selected areas of the patterns). The Bayesian network also significantly outperformed two multivariate statistical techniques (linear discriminant analysis and classification trees) and proved to be less sensitive to the size of the training set and more robust in the response to patterns belonging to unknown species.

Objective: To describe the clinical findings in 52 racehorses with axial deviation of the aryepiglottic folds (ADAF) and to report outcome in 33 of these horses after either rest or transendoscopic laser excision of aryepiglottic fold tissue., Study Design: Retrospective study., Animal or Sample Population: Racehorses admitted for high-speed treadmill (HST) evaluation of poor performance., Methods: Medical records and videotapes of resting and exercising videoendoscopic examinations were reviewed. Racing performance records and owner or trainer interviews, at least 1 year after HST examination, were used to compare results after either surgical management or rest in 33 horses with ADAF and no other upper-airway abnormalities., Results: ADAF occurred in 6% of horses evaluated for poor performance. No breed or gender predisposition existed, but horses with ADAF were younger than the overall population evaluated on the HST. Of 52 horses with ADAF, 19 horses had at least one other upper-airway abnormality. There was no apparent association between ADAF and other causes of dynamic upper-respiratory obstruction. Surgical correction was successfully performed in standing or anesthetized horses without complications. When ADAF was the only upper-airway obstruction, 75% of horses that had surgery and 50% of rested horses had objective improvement in performance. Owners and trainers also perceived greater improvement in performance in horses that had surgery., Conclusions: Whereas surgical management of ADAF is recommended, clinical experience indicated that it is not required to resolve ADAF in all horses. However, owners and trainers of horses that had surgery were more satisfied with outcome than those with horses managed conservatively., Clinical Relevance: Diagnosis of ADAF can only be made by videoendoscopic evaluation during high-speed exercise. Transendoscopic laser excision of the collapsing portion of the aryepiglottic folds can be performed safely in standing horses and results in resolution of airway obstruction and rapid return to training., (Copyright 2001 by The American College of Veterinary Surgeons.)

Objective: To determine results for horses undergoing a high-speed treadmill examination, including videoendoscopy of the pharynx and larynx before and during exercise, echocardiography before and after exercise, and electrocardiography before, during, and after exercise, because of poor performance., Design: Retrospective study., Animals: 348 horses., Results: A definitive diagnosis was obtained for 256 (73.5%) horses. One hundred forty-eight horses had dynamic obstruction of the airway during exercise, 33 had clinically important cardiac arrhythmias alone, 22 had a combination of dynamic airway obstruction and clinically important cardiac arrhythmias, 19 had poor cardiac fractional shortening immediately after exercise, 10 had exertional rhabdomyolyis, 15 had clinically apparent lameness, and 9 had other disorders. Thirty-nine of the horses with dynamic obstruction of the airway during exercise had multiple airway abnormalities. Fifty-three horses also had subclinical myopathy, Conclusions and Clinical Relevance: Results suggest that a complete evaluation, including a high-speed treadmill examination, should be conducted in horses with poor performance, regardless or whether horses do or do not have a history of abnormal respiratory noises and particularly if the horses have grade-II or -III left laryngeal hemiplegia.

Bacteriocins produced by lactic acid bacteria are a heterogeneous group of peptide inhibitors which include lantibiotics (class I, e.g. nisin), small heat-stable peptides (class II, e.g. pediocin AcH/PA1) and large heat-labile proteins (class III, e.g. helveticin J). Many bacteriocins belonging to the first two groups can be successfully used to inhibit undesirable microorganisms in foods, but only nisin is produced industrially and is licensed for use as a food preservative in a partially purified form. This review focuses on the production and purification of class I and class II bacteriocins from lactic acid bacteria. Bacteriocin production is growth associated but the yield of bacteriocin per unit biomass is affected by several factors, including the producing strain, media (carbohydrate and nitrogen sources, cations, etc.) and fermentation conditions (pH, temperature, agitation, aeration and dilution rate in continuous fermentations). Continuous fermentation processes with cell recycle or immobilized cells can result in a dramatic improvement in productivity over batch fermentations. Several simple recovery processes, based on adsorbing bacteriocin on resins or silica compounds, have been developed and can be used to build integrated production processes.

Objective: To evaluate results of cytologic examination of specimens obtained by means of tracheal washes (TW) in 42 horses with a history of poor performance., Design: Cross-sectional case series., Animals: 42 horses with a history of poor performance., Procedure: A TW was performed via endoscopy before and after horses exercised on a high-speed treadmill, and specimens were evaluated microscopically and graded., Results: Ten (24%) horses were considered to be clinically normal before and after exercise. Pulmonary hemorrhage was diagnosed in 8 (19%) horses. One horse had evidence of exercise-induced pulmonary hemorrhage (EIPH) before exercise and pulmonary hemorrhage and allergic or inflammatory airway disease (IAD) after exercise. Five (12%) horses had IAD, and 1 had IAD and pulmonary hemorrhage after exercise. Seven (17%) horses had evidence of EIPH and IAD in both specimens. Four (10%) horses with EIPH had an increase in the proportion of hemosiderophages in the specimen obtained after exercise. Specimens obtained before exercise in 6 (14%) horses were not representative of the respiratory tract and could not be compared with specimens obtained after exercise., Clinical Implications: Interpretation of TW specimens obtained before and after exercise differed for only 5 of 36 (14%) horses. Specimens obtained after exercise were more likely to reveal airway disease. All specimens obtained after exercise adequately represented the respiratory tract, whereas 6 specimens obtained before exercise did not. Specimens obtained after exercise contained more airway secretions and had less cytologic evidence of pharyngeal contamination. Therefore, we recommend that TW samples be obtained after exercise in horses.

Objective: To evaluate laryngeal function by means of videoendoscopy during high-speed treadmill exercise in racehorses with grade-III left laryngeal hemiparesis at rest and to determine outcome of treatment., Design: Retrospective study., Animals: 26 racehorses., Procedure: Videoendoscopy of the larynx was performed while horses were at rest and exercising on a treadmill. Horses were classified as having grade-IIIA, -IIIB, or -IIIC laryngeal hemiparesis on the basis of the degree of arytenoid cartilage abduction maintained during exercise. Postoperative racing performance was determined by evaluating race records and conducting telephone surveys., Results: 20 (77%) horses had grade-IIIC laryngeal hemiparesis (i.e., severe dynamic laryngeal collapse during exercise). Eighteen underwent surgery, and racing performance was improved in 9. Five (19%) horses had grade-IIIB laryngeal hemiparesis (i.e., left arytenoid cartilage and vocal fold were maintained in an incompletely abducted position during exercise). Four underwent surgery, and racing performance was improved in 1. One (4%) horse had grade-IIIA laryngeal hemiparesis (i.e., full abduction of arytenoid cartilage during exercise); surgery was not performed., Clinical Implications: Videoendoscopy is useful in determining dynamic laryngeal function in racehorses with grade-III laryngeal hemiparesis at rest.

The many testing methods available and the increasing sophistication of diagnostic equipment have enhanced greatly our capabilities to diagnose causes of exercise intolerance in the equine athlete during the last several years. High-speed treadmill examination has become the focus of this form of evaluation. Not all clinicians perform or have access to high-speed treadmill examinations. Testing methods that require the use of the high-speed treadmill, as well as methods that do not, are discussed.

Six forelimb specimens from three adult horses had the fetlock joint fused by application of a dorsal plate and by a screw placed in lag fashion through the metacarpus to each proximal sesamoid bone. Five specimens were instrumented on the central dorsal surface of the plate with a single rosette strain gage, and the plate of the sixth specimen was instrumented with four longitudinally oriented single-axis strain gages. The specimens were loaded axially in compression to 4,000 N in a cast (test 1), in a cast with a heel block (test 2), and uncast (test 3). The principal angle of strain in all specimens, in all tests, closely approximated the vertical axis at loads < 1,000 N. The principal angle in uncast specimens was significantly different at loads > 1,000 N than the cast specimens (P < .05). At loads > 3,000 N, the principal angle in test 3 closely approximated the horizontal axis, indicating a change from tension to compression on the dorsal surface of the plate, whereas the principal angle of the cast specimens was unchanged. Specimens in a cast (tests 1 and 2) suffered less surface deformation than did uncast specimens (test 3). Therefore, the cast changed the direction and extent of bending at the point of fixation, and thereby decreased the deformation of the plate. This effect would lead to greater fatigue life of the implant in the cast specimens compared with the uncast specimens.

Keratan sulfate (KS) is a glycosaminoglycan, distribution of which is confined mostly to hyaline cartilage. As such, it is a putative marker of hyaline cartilage catabolism. In experiment 1, a focal osteochondral defect was made arthroscopically in 1 radial carpal bone of 2 ponies, and in 2 other ponies, chymopapain was injected into the radiocarpal joint to induce cartilage catabolism. Sequential and concurrent plasma and synovial fluid concentrations of KS were measured, up to 13 months after induction of cartilage injury, to determine whether changes in KS concentrations reflected cartilage catabolism. In experiment 2, a large, bilateral osteochondral defect was made in the radial carpal bones of 18 ponies, which were subsequently given postoperative exercise and/or injected intra-articularly with 250 mg of polysulfated glycosaminoglycan (PSGAG). Medication was given at surgery, then weekly for 4 weeks. Blood samples were collected and synovial fluid was aspirated before surgery, when medication was given, and at postmortem examination (postoperative week 17). The KS concentration was measured in these fluids to determine whether changes in KS concentration indicated an effect of joint treatment. In experiment 1, the concentration of KS in synovial fluid was highest 1 day after joint injury, and the concentration in plasma peaked 2 days after joint injury. For ponies receiving chymopapain intra-articularly (generalized cartilage catabolism), a fivefold increase over baseline was observed in the concentration of KS in plasma (peak mean, 1.2 micrograms/ml), and a tenfold increase over baseline in synovial fluid (peak mean, 2.0 mg/ml) was observed. On average, these maxima were threefold higher than values in fluids of ponies with osteochondral defects (focal cartilage disease). In experiment 2, nonexercised ponies had lower KS concentration (as a percentage of the preoperative concentration) in synovial fluid than did exercised ponies at all postoperative times, and at postoperative week 17, this effect was significant (P < 0.05). This may be related to decreased turnover of KS in articular cartilage attributable to stall confinement and late increase in turnover related to exercise. Seventeen weeks after surgery, synovial fluid from exercised, medicated ponies had significantly (P < 0.05) higher KS content than did fluid from exercised, nomedicated ponies. This indicated that exercise, when combined with medication, may increase KS release from articular cartilage. Synovial fluid from medicated joints of nonexercised ponies had significantly (P < 0.05) lower KS concentration than did synovial fluid from nonmedicated joints of nonexercised ponies.(ABSTRACT TRUNCATED AT 250 WORDS)

Medical records of 57 horses admitted between 1980 and 1991 because of basal sesamoidean fractures were evaluated. Radiographic measurements of fragment size and fracture characteristics were recorded to determine their relationship to outcome. A successful outcome was assessed on the basis of the ability to return to racing, ability to race more than one time, and ability to finish first, second, or third. Any change in racing class also was assessed. There was a significant (P < 0.001) overrepresentation of Thoroughbreds, compared with other breeds in the hospital population. Fractures of the forelimbs accounted for 50 of the 57 fractures, and the right front medial sesamoid was affected significantly (P < 0.0001) more frequently than other proximal sesamoids. Fifty-nine percent of the horses returned to race at least 1 time regardless of treatment, and 41% finished first, second, or third. Horses with smaller fragments (shorter dorsopalmar length) tended to do better than horses with larger fragments. Horses without comminuted fractures tended to do better than horses with comminuted fractures, and horses with fragments only mildly (< 3 mm) displaced had significantly (P < 0.05) better outcomes than did horses with severe displacement of fragments. Only 19% of the horses with moderate (> 3 mm) displacement of fragments raced more than once, whereas 63% of horses with mild displacement of fragments returned to race more than once. Seventy-three percent of the horses that had the fragment removed surgically returned to race, and 57% dropped in class. Only 48% of the horses that did not have the fragment removed returned to race, and 87% dropped in class.(ABSTRACT TRUNCATED AT 250 WORDS)

Enterococcus faecium DPC 1146 produces a bacteriocin, enterocin 1146, which is inhibitory to Listeria monocytogenes . Enterocin 1146 was produced in GM17 and in milk. The bacteriocin was partially purified by ammonium sulfate precipitation. Its molecular weight, estimated by SDS-PAGE, was 3.0 kDa. It could be stored at -20°C without loss of activity, but pH had a marked effect on enterocin 1146, which was more stable at both high (up to 120°C) and low temperatures (4°C) at pH 5 than at pH 7 and 9. The sensitivity of 57 strains belonging to 35 different species was studied using a critical dilution assay. L. monocytogenes and L. innocua were most sensitive; enterocin 1146 had a bactericidal effect on Listeria . Starter and nonstarter lactic acid bacteria (except Lactobacillus sake )were insensitive or relatively resistant to the bacteriocin. Genetic determinants for bacteriocin production and immunity do not appear to be plasmid borne.

Enterocin 1146, a bacteriocin produced by Enterococcus faecium . DPC1146, has a rapid bactericidal effect on Listeria in buffer systems, broth, and milk. In trypticase soy broth, increasing the bacteriocin dose and/or decreasing the pH extended the lag phase of Listeria innocua . A logarithmic relationship was found between response (as proportion of survivors or growth compared to a control) and dose. Increasing the inoculum level of the indicator reduced the effectiveness of enterocin 1146. Log-phase cells of L. innocua were more resistant than stationary-phase cells in both broth and buffer systems. In milk treated with 250 arbitrary units of enterocin 1146/ml and inoculated with 10 3 or 10 5 CFU/ml of Listeria monocytogenes Scott A, populations reached only 5-14% of the control after 24 h at 30°C, with numbers exceeding 10 7 , while at 6°C a slow decrease in population was found.

Using isocratic retention parameters, the gradient elution retention time for several proteins has been calculated. The gradient retention time calculation is based on fitting the isocratic retention data to an equation of the form: log k' = m log (1/[Ca2+]) + log K and on applying well-established principles of gradient elution. A good correlation between the observed and calculated retention times for several test proteins was obtained at various total gradient times and column flow-rates. Conversely, isocratic retention parameters characterizing protein retention can be calculated from gradient elution retention data. However, even with retention data of high quality, small errors are amplified by the log-log nature of the ion-exchange isocratic retention model employed. Based on the close correlation between predicted and observed gradient retention times, no evidence for protein denaturation resulting from immobilization of the protein at high initial k' values at or near the column inlet was observed.

In the golden hamster, mating with an intact male results in both a reduction in receptivity shown toward a second male (short-term effect) as well as an absolute abbreviation of the receptive period (long-term effect). The present study examined the components of the mating situation responsible for the long-term effect. In experiment 1 we showed that the paradigm used to determine receptivity (i.e., brief hourly exposures to an intact male) did not affect the duration of receptivity whereas an ad lib mating early during the receptive period resulted in a 40% reduction in the length of behavioral estrus. In experiment 2 females received various types of mating stimuli, e.g., mounts from an apronned male, intromissions from a vasectomized male, a simulated copulatory plug, manually-induced lordosis, or several of these stimuli in combination. Only intromissions from an intact or vasectomized male significantly reduced the duration of behavioral estrus. Although as few as 10 intromissions from a vasectomized male were effective in significantly shortening the receptive period, higher levels of vaginocervical stimulation from a single male (experiment 3) or from additional males (experiment 4) effected significantly greater reductions in the length of behavioral estrus. Our results further demonstrated that only those females that subsequently became pseudopregnant showed significant abbreviations of behavioral estrus. The association of these two phenomena is of obvious adaptive value since it provides a mechanism to terminate a behavior once its function has been achieved.

The effects of urea on the high-performance cation-exchange chromatography of hen egg lysozyme are reported. The capacity factor, k', has been determined as a function of cation concentration with a polyaspartate column using the acetates of Na+, K+, Ca2+ and Mg2+. Urea decreases lysozyme retention. Plots of log k' vs. log ionic strength show linear relationships. The slope of the plot describing the Ca2+ elution of lysozyme was the same in the presence of 5 M urea as in its absence. In strong urea solutions and at elevated temperatures, lysozyme denaturation is evidenced by a marked decrease in k'. The temperature range for denaturation corresponded closely to that observed in intrinsic fluorescence and circular dichroism measurements. The potential utility and limitations of high-performance ion-exchange chromatography for studying protein denaturation are discussed.

Retention parameters of alpha-chymotrypsinogen-A were determined by isocratic elution for a series of concentrations of calcium acetate and sodium acetate both in the presence and absence of urea. Under non-denaturing conditions of temperature and urea concentration, urea facilitated elution. Under reversible denaturing conditions a sharp drop in chromatographic retention was observed over a narrow temperature range which could be correlated with equilibrium measurements of the protein fluorescence. Retention of both native and denatured protein could be fit to a non-mechanistic retention model by plotting the log k' against log salt concentration. Conventional interpretation of these findings indicates that, while the number of ions displaced during binding is greater for the denatured protein, the affinity per ion decreases since the retention of denatured protein is much less than native. Elution profiles obtained under partially denaturing conditions showed a strong flow-rate dependence. We attribute these observations to a rate of equilibration between native and denatured protein that is on the timescale of the chromatographic rate processes.

After 12-15 ejaculatory series, each consisting of several short intromissions (2- to 3-s vaginal penetration) and an ejaculation, male golden hamsters adopt an altered copulatory pattern consisting of long intromissions (5- to 25-s penetration with intravaginal thrusting). Receptivity declines and the tendency for the female to attack the male increases at about the time of this shift in copulatory pattern. Because the mean interintromission interval (III) between short intromissions is about 8 s compared to 100 s between long intromissions, it is possible that females detect this difference and adjust their mating accordingly. When the III between short intromissions from a rested male was experimentally increased to 100 s by use of a halter and lead device, the duration of lordosis was significantly less than that displayed by females paired with control males (8-s III) and virtually the same as that displayed by females paired with males that produced only long intromissions. This suggests that the female uses the temporal patterning of intravaginal stimulation as one criterion for terminating mating with a particular male.

A rapid, precise, and reproducible automated high pressure liquid chromatographic method was developed to determine xylitol in the presence of mannitol and sorbitol in chewing gums and confectionery products. A mobile phase of water and methanol elutes the polyols simultaneously from a cation-exchange column without pretreatment or derivatization. Injections into the liquid chromatograph were made by an autosampler, and date reduction was performed with a programmable electronic integrator. Average recoveries for one level each of mannitol, sorbitol, and xylitol in 6 replicate samples prepared by the standard addition technique were 102.5, 100.5, and 100.7%, respectively.