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c.m. logue, j.j. sheridan and d. harrington. 2004.This study was carried out to determine the survival ofEscherichia coliO157:H7 and subsequent shelf life of beef subjected to subatmospheric steam at differing temperatures.A specifically built, laboratory scale decontamination apparatus was used in decontamination trials to examine the effect of condensing steam at differing subatmospheric pressures on the survival ofE. coliO157:H7 on meat. Beef slices were inoculated with a nontoxigenicE. coliO157:H7 strain and subjected to condensing steam at temperatures of 55, 65 and 75°C. Following treatment, the decontaminated meat was packaged and stored in air or under vacuum at temperatures of 10 or 0°C for up to 42 days. Microbiological analysis of the decontaminated and a control product (not subjected to any heat treatment) was carried out at regular intervals over the storage time of the product. Overall, significant reductions (ca1·5 log10 CFU cm−2) in pathogen numbers were observed at a steam treatment temperature of 75°C, however, postprocess storage conditions were important in ensuring no re-growth of the pathogen and this was best achieved by storage under vacuum at 0°C.Steam had a significant impact in reducingE. coliO157:H7 populations, but storage conditions post-treatment were important for ensuring inhibition of the pathogen.This study indicated that subatmospheric steam could have significant application in the decontamination of meat primals postfabrication, immediately prior to packaging thus ensuring a safer product for consumers. [ABSTRACT FROM AUTHOR]

Aims: The aim of this research was to examine the effects of preslaughter washing, pre-evisceration washing, final carcass washing and chilling on final carcass quality and to evaluate these operations as possible critical control points (CCPs) within a pork slaughter hazard analysis and critical control point (HACCP) system. Methods and Results: This study estimated bacterial numbers (total viable counts) and the incidence of Salmonella at three surface locations (ham, belly and neck) on 60 animals/carcasses processed through a small commercial pork abattoir (80 pigs d-1 ). Significant reductions (P < 0·05) in bacterial numbers were noted at some stages of the slaughter/dressing process, i.e. the process of hair removal (scalding–dehairing and singeing) resulted in an approx. 4·5 log10 cfu cm-2 decrease in bacterial numbers. A significant increase (P < 0·05) in bacterial numbers was observed after pre-evisceration washing. Final washing increased the bacterial counts to between 3·6 and 3·8 log10 cfu cm-2 while chilling effected a small but statistically significant (P < 0·05) increase to between 4·5 and 4·7 log10 cfu cm-2 . The incidence of Salmonella on pigs at the farm was 27%, decreasing to 10% after preslaughter washing. However, stunning and bleeding effected a considerable increase in Salmonella contamination and the incidence after these operations was 50%, which was reduced to 0% during the scalding–dehairing process. Conclusions: Washing the live animals and subsequent carcasses with cold water is not an effective control measure but chilling may be used as a CCP. Significance and Impact of the Study: Recent changes in European Union legislation legally mandate HACCP in pork slaughter plants. This research will provide a sound scientific basis on which to develop and implement effective HACCP in pork abattoirs. [ABSTRACT FROM AUTHOR]

Aims To establish the extent, if any, of peripheral changes in asymptomatic patients with severe left ventricular dysfunction.Methods and Results Nine asymptomatic and nine symptomatic patients with left ventricular ejection fraction, <25%, matched for age and left ventricular ejection fraction (asymptomatic vs symptomatic, age: 52±1·5 vs 55·9±2·5 years [Mean±SEM], left ventricular ejection fraction: 16±2 vs 19±2%P=0·23 and 0·48, respectively) were studied and compared with 26 age-matched normal controls. We assessed exercise capacity, leg blood flow (occlusion plethysmography), respiratory muscle strength, quadriceps maximal isometric strength, fatigue and CT cross-sectional muscle area at mid thigh. Fatigue was expressed as the percentage reduction in maximal strength following a 20min fatiguing protocol. There was a graded increase in peak oxygen consumption comparing symptomatic, asymptomatic and control groups (16·6±1·3 vs 27·1±1·6 vs 32·8±1·3ml.min−1.kg−1respectively, ANOVA P<0·0001). Between the three groups there was significant variation in muscle strength (P<0·0001), endurance (P=0·0002) and cross-sectional area (P=0·0003) and in peak blood flow (P=0·027) and respiratory muscle strength (P<0·05). When asymptomatic patients and controls were compared no significant differences existed.Conclusions Patients with severe left ventricular dysfunction may have near normal exercise capacity and no peripheral changes. Exercise capacity may depend less upon left ventricular function than on the presence or absence of peripheral factors. [ABSTRACT FROM PUBLISHER]

Examines the inactivation of Listeria monocytogenes and Yersinia enterocolitica in minced beef. Use of vacutainers for heating the beef; Effects of pre-heating process of beef on the D[sub 60]-values; Duration of the heating-up phase in laboratory experiments on the subsequent D-values.

AimsTo investigate the frequency of pathogenic mitochondrial DNA mutations in idiopathic cardiomyopathy.Methods and ResultsWe investigated the occurrence of seven previously reported pathogenic mitochondrial DNA point mutations in 52 patients with idiopathic dilated cardiomyopathy (blood n=33, myocardium n=19), 10 patients with hypertrophic cardiomyopathy (blood n=7, myocardium n=3), 67 controls with ischaemic heart disease (blood n=53, myocardium n=14) and eight controls with no overt cardiac disease (blood n=4, myocardium n=4). Total DNA or cell lysates were studied by polymerase chain reaction amplification and restriction fragment length polymorphism analysis for the identification of the following mitochondrial DNA point mutations: A3243G, A3252G, A3260G, A4269G, A8344G, T8993G/C and T9997C. None of these point mutations were detected in the blood or myocardium of any of the individuals with dilated or hypertrophic cardiomyopathy or in the controls. In addition we investigated the occurrence of major deletions of mitochondrial DNA in eight patients with dilated cardiomyopathy (myocardium n=7, skeletal muscle n=1), three patients with ischaemic heart disease (myocardium n=3) and one control myocardium by Southern blot analysis. Deletions were not detected in any of the patients.ConclusionThe results suggest that although these mutations are known to be associated with specific cardio-myopathies, they are not a common feature of idiopathic cardiomyopathy.The European Society of Cardiology [ABSTRACT FROM PUBLISHER]

Aims The peripheral chemoreflex may be augmented in chronic heart failure and may play a role in its pathophysiology including the mediation of exercise hyperpnoea and sympathetic activation. The objective of this study was to characterize the patients with an augmented peripheral chemoreflex. Methods and results Peripheral chemoreflex sensitivity was assessed by measuring the ventilatory response to hypoxia using transient inhalations of pure nitrogen in 50 patients with chronic heart failure (age 58·±12·1 (SD) years; radionuclide left ventricular ejection fraction 26·5±13·0%). The peripheral chemoreflex of 12 healthy controls with similar demographic characteristics was 0·272±0·201 1.min−1. %Sao2−1 compared with 0·673±0·4101.min−1.%Sao2−1 (P<0·0001) in the chronic heart failure patients. Using 2 standard deviations above the mean level of the controls' peripheral chemoreflex sensitivity as the upper limit of normal, we defined an augmented chemoreflex as greater than 0·6751.min−1.%Sao2−1 Twenty of the chronic heart failure patients (40%) demonstrated such an augmented peripheral chemoreflex. Compared with patients with peripheral chemoreflex sensitivity within the normal range, they had a reduced peak oxygen consumption during cardiopulmonary exercise (15·1±4·4 vs 18·5±5·8 ml.kg−1.min−1, P=0·02), reduced radionuclide left ventricular ejection fraction (21·8±11·8 vs 29·4±13·1%, P=0·046) and were in a worse New York Heart Association functional class (2·8 vs 2·4 P=0·05). The ventilatory response to exercise, as characterized by the regression slope relating minute ventilation to carbon dioxide output during exercise, was also higher (40·48±9·32 vs 34·54±7·19, P=0·02), consistent with the role of the peripheral chemoreflex in mediating exercise hyperpnoea. There was also an increased proportion of patients with non-sustained ventricular tachycardia in the group with an augmented peripheral chemoreflex (61% vs 21%, chisquared 7·08, P<0·01). No difference was seen in the age, height, weight and lung function measurements of these patients compared with the normal chemoreflex group. Conclusion An augmented peripheral chemoreflex is a common finding in chronic heart failure patients, one associated with increasing severity and with the exercise hyperpnoea seen in the condition. That there was an excess of patients with non-sustained ventricular tachycardia in the group with an augmented peripheral chemoreflex may be related to the chemoreflex-driven sympathetic stimulation. The peripheral chemoreflex may be important in the pathophysiology of chronic heart failure, both in terms of symptoms and exercise limitation. [ABSTRACT FROM PUBLISHER]

A full likelihood method is proposed to analyse continuous longitudinal data with nonignorable (informative) missing values and non-monotone patterns. The problem arose in a breast cancer clinical trial where repeated assessments of quality of life were collected: patients rated their coping ability during and after treatment. We allow the missingness probabilities to depend on unobserved responses, and we use a multivariate normal model for the outcomes. A first-order Markov dependence structure for the responses is a natural choice and facilitates the construction of the likelihood; estimates are obtained via the Nelder-Mead simplex algorithm. Computations are difficult and become intractable with more than three or four assessments. Applying the method to the quality-of-life data results in easily interpretable estimates, confirms the suspicion that the data are non-ignorably missing and highlights the likely bias of standard methods. Although treatment comparisons are not affected here, the methods are useful for obtaining unbiased means and estimating trends over time. [ABSTRACT FROM AUTHOR]

Administration of high-dose (250 mg/kg) cyclophosphamide (CY) to guinea-pigs and mice 3 days prior lo immunization with inactivated vaccines derived from Venezuelan encephalitis virus (VE). Coxiella burnetii and Francisella tularensis resulted in accentuated and prolonged delayed-type hypersensitivity (DTH) and in vitro cellular immunity (CMI) to specific antigen. Humoral antibodies were either absent or significantly lower in CY-pretreated animals compared to immunized non-pretreated controls. CY-pretreatments precluded protection in the VE virus model, suggesting that resistance is related to antibody. In the Q fever model, the protective immunogenicity of vaccine was preserved or increased by CY pretreatment suggesting that cell-mediated immunity is the important factor. In the tularaemia bacterial system, there was a complex effect of CY pretreatment on the low-grade protection afforded by killed vaccine against virulent infection. These findings suggest that the inability of killed vaccines to induce high-grade resistance against tularaemia and Q fever may be due in part to a suppressive B cell response which is eliminated by CY. These studies have given useful information on the relative significance of components of the specific immune response and may lead to an increased understanding of the mechanisms of action of vaccines and adjuvants. [ABSTRACT FROM AUTHOR]

Successful cytogenetic studies were performed on 69 biopsies from 64 patients with non-Hodgkin's lymphoma bearing a t(14;18)(q32;q21) translocation. This translocation appears to be a primary abnormality associated with the development of certain B-cell non-Hodgkin's lymphomas. We correlated the occurrence of secondary abnormalities, in addition to the t(14;18)(q32;q21), with histologic subtype to test the hypothesis that secondary abnormalities correlate with more aggressive histologic appearance. A large number of secondary abnormalities were identified, the most frequent being additional copies of chromosomes 7 (30%), 12 (22%), 18 (22%), 20 (16%), or 21 (14%), deletion of a portion of the long arm of chromosome 6 (17%), and either an additional chromosome 17 or an isochromosome for the long arm of chromosome 17 (13%). An extra chromosome 7 was highly associated with a diffuse histologic pattern; it was present in 52% of patients with a diffuse pattern and in only 15% of those with a follicular pattern ( = .002). A weaker association with a diffuse growth pattern was found for the addition of chromosome 17 or an i(17q); it was found in 24% of patients with a diffuse pattern and only 5% of those with a follicular pattern ( = .05). No other significant correlations between secondary chromosome abnormalities and histologic subtype were identified. Although the explanation for this association is not clear, it appears that patients with B-cell non-Hodgkin's lymphomas bearing the t(14;18) (q32;q21) translocation which also have an additional chromosome 7 are likely to exhibit a diffuse growth pattern. [ABSTRACT FROM PUBLISHER]

Erectile dysfunction (ED) affects ~50% of men aged 40-70 and has a high impact on men's health. We examine the mechanism of how apoptosis occurs in ED patients and in a CN injury rat model, to determine points of apoptosis intervention for therapy development. Immunohistochemical analysis and western analysis for caspase 3 cleaved, -8 and -9 (pro and active forms) were performed in corpora cavernosal tissue from Peyronie's, prostatectomy and diabetic ED patients (n=56), and in penis from adult Sprague Dawley rats that underwent CN crush and were sacrificed after 1-9 days (n=16). [Extracted from the article]

We examine parallel mechanisms of how apoptosis occurs in response to SHH inhibition and in a CN injury rat model in order to identify significant points for intervention and ED therapy development. Immunohistochemical analysis and western analysis for caspase 3 cleaved, -8 and -9 (pro and active forms) were performed in corpora cavernosal tissue from adult Sprague Dawley rats that underwent SHH inhibition in the corpora cavernosa and in the pelvic ganglia (PG), and after CN crush with SHH treatment (n=49). Understanding how apoptosis takes place after CN injury and in response to SHH treatment is critical for translation to ED patients. [Extracted from the article]

We developed a self-assembling peptide amphiphile (PA) nanofiber hydrogel for extended release of SHH protein to the penis after CN injury, to suppress SM apoptosis. SM was 48% higher with SHH treatment, and doubling the concentration of SHH resulted in higher SM preservation (76%). Proliferation of SM and endothelium also occur in the corpora cavernosa after CN injury, and this response is increased with SHH PA treatment. [Extracted from the article]

Introduction: A key aspect of erectile dysfunction (ED) development is loss of innervation to the penis. The cavernous nerve (CN) is frequently damaged in prostatectomy and diabetic patients with ED, initiating changes in penile morphology including an acute and intense phase of apoptosis in penile smooth muscle followed by increasing collagen and fibrosis. This remodeling process alters penile architecture so that there is less smooth muscle in the corpora cavernosa, and what remains is less compliant/able to relax in response to normal neurotransmitter signaling, and ED results. Thus, novel therapies are needed which target the underlying remodeling process. We have shown previously that Sonic hedgehog (SHH) is a critical regulator of penile smooth muscle, and that SHH protein treatment suppresses penile remodeling after CN injury through an unknown mechanism. Objective: We examine if part of the mechanism of how SHH preserves smooth muscle after CN injury involves bone morphogenetic protein 4 (BMP4) and GREMLIN (GREM1). Methods: Primary cultures of smooth muscle cells were established from patients who had prostatectomy, diabetes, hypertension and Peyronie's (control, n=18). Cultures were characterized by immunohistochemical analysis (IHC) for ACTA2, CD31, P4HB and nNOS. Smooth muscle cell growth was quantified in response to BMP4 and GREM1. Adult Sprague Dawley rats that were uninjured or underwent CN crush injury, were treated with BMP4, GREM1 or MSA (control) proteins via Affi-Gel beads (n=16) or peptide amphiphile (PA, n=26) for 3 and 14 days, and trichrome stain was performed. Sprague Dawley rats underwent CN injury (n=9) and SHH PA treatment for 1, 2 and 4 days (n=9) and western was performed assaying for BMP4 and GREM1 proteins in comparison to sham (n=3) controls. Adult Sprague Dawley rats were treated with 5E1 SHH inhibitor (n=6) or IgG (control, n=6) for 2 and 4 days, and BMP4 and GREM1 localization were examined by IHC. Statistics were performed by ANOVA with Scheffe's posthoc test. Results: Patient primary smooth muscle cultures were established. BMP-4 increased patient smooth muscle cell growth and GREM1 decreased growth. In rats BMP4 treatment via Aff-Gel beads and PA increased smooth muscle growth at 3 and 14 days of treatment. GREM1 treatment caused collagen and smooth muscle growth at 3 days, which switched to primarily collagen at 14 days. CN injury increased BMP4 and GREM1. SHH PA altered western band size suggesting alternative cleavage and range of signaling. SHH inhibition in rats changed BMP4 and GREM1 localization. Conclusions: BMP4 treatment increases smooth muscle in patient cultures and in our rat model. BMP4 and GREM1 mediate a switch from smooth muscle to collagen induction. SHH treatment alters BMP4 and GREM1 localization and range of signaling, which can affect penile morphology. Whether GREM1 directly effects collagen induction or acts strictly through inhibition of BMP4 is unknown and requires further study. Part of the mechanism of how SHH regulates corpora cavernosal smooth muscle, involves BMP4 and GREM1. Understanding how SHH PA impacts penile morphology after CN injury will aid development of ED therapies. Disclosure: No. [ABSTRACT FROM AUTHOR]

Introduction: Cavernous nerve (CN) injury caused by prostatectomy and diabetes, initiates a remodeling process (smooth muscle apoptosis and increased collagen) in the corpora cavernosa of the penis of patients and animal models that is an underlying cause of erectile dysfunction (ED). The Sonic hedgehog (SHH) pathway plays an essential role in the response of the penis to denervation, as collagen increases with SHH inhibition and decreases with SHH treatment. Part of the mechanism of how SHH impacts collagen may involve bone morphogenetic protein four (BMP4) and GREMLIN (GREM1). Microarray analysis identified increased GREM1 (BMP4 antagonist) in corpora cavernosa of ED patients and SHH is a regulator of GREM1 in the limb bud. Objective: We will examine the relationship between SHH, BMP4, GREM1 and collagen in penis of ED patients and rat models of CN injury, SHH inhibition, and SHH, BMP4 and GREM1 treatment. Methods: Corpora cavernosa of Peyronie's (control), prostatectomy and diabetic ED patients was obtained (n=26). Adult Sprague Dawley rats (n=90) underwent either: 1. CN crush (1-7 days) or sham control, 2. CN injury and BMP4, GREM1 or MSA (control) protein treatment via Affi-Gel beads or peptide amphiphile (PA) for 14 days, 3. 5E1 SHH inhibitor, IgG and PBS (controls), or SHH protein treatment for 2-4 days. Immunohistochemical and western analysis for BMP-4 and GREM1, and collagen analysis by hydroxyproline and trichrome stain were performed. Results: BMP4 and GREM1 proteins were identified in corpora cavernosal smooth muscle of prostatectomy, diabetic and Peyronie's patients, and in rat penis, sympathetic nerve fibers, perineurium, blood vessels, and urethra. Collagen decreased 25.4% in rats with CN injury and BMP4 PA treatment (p=0.02) and increased 61.3% with CN injury and GREM1 treatment (p=0.005). Trichrome showed increased collagen in rats treated with GREM1. BMP4 increased smooth muscle. Western identified increased BMP4 and GREM1 in corpora cavernosa of prostatectomy and diabetic patients, and after CN injury (1-2 days) in our rat model. Localization of BMP4 and GREM1 changed with SHH inhibition. SHH treatment increased BMP4 and GREM1. Aging did not impact BMP4 and GREM1 localization. Conclusions: BMP4 and GREM1 are downstream targets of SHH that impact collagen and may be useful in collaboration with SHH to prevent penile remodeling and ED. Better understanding of penile remodeling and how fibrosis occurs is essential for development of novel therapies for ED. Disclosure: No. [ABSTRACT FROM AUTHOR]

Erectile dysfunction (ED) treatments are minimally effective in prostatectomy and diabetic patients due to injury to the cavernous nerve. With denervation the critical smooth muscle undergoes apoptosis and the penis becomes fibrotic, thus altering corpora cavernosal architecture. In order to devise novel ED therapies, prevention of corpora cavernosal remodeling is critical. Sonic hedgehog (SHH) treatment by peptide amphiphile (PA) nanofiber hydrogel, suppresses smooth muscle apoptosis and improves erectile function in a rat CN injury ED model. We examine if human corpora cavernosal smooth muscle responds in a similar manner to SHH treatment. In this study we: 1.) Examine if human corpora cavernosal smooth muscle cells from primary culture of ED patient tissues, respond to Sonic hedgehog (SHH) treatment by increasing growth rate, as in our in vivo animal model. 2.) Determine if human corpora cavernosal cells from patients with differing underlying mechanisms of ED development (prostatectomy, diabetes, hypertension, cardiovascular disease), respond similarly to SHH treatment. 3.) Identify if organ culture conditions (glucose) affect growth of corpora cavernosal smooth muscle and the response to SHH signaling. 4.) Examine the vascular component of ED (hypertension and cardiovascular disease). Human corpora cavernosal tissue was obtained from prostatectomy, diabetic, hypertension, cardiovascular disease and Peyronie's (control) patients (n=40) that were under going prosthesis implant to treat ED. Primary cultures (n=17) were established, and corpora cavernosal cells from passage 3-4 were treated with SHH protein, MSA (control), 5E1 SHH inhibitor, and PBS (control). Growth was quantified by counting the number of cells using a hemocytometer at 3-4 days. The concentration of SHH protein for maximal growth was examined, in addition to a lipid modified more active SHH peptide. SHH treatment increased smooth muscle growth in human corpora cavernosal cells from prostatectomy, diabetic, and Peyronie's patients in a similar manner (43-53%). SHH inhibition decreased smooth muscle growth (24-32%). There was no difference in growth using 25ug and 10ug SHH peptide. A more active (150X) SHH peptide further enhanced growth (20%). SHH protein increased growth more in diabetic smooth muscle under high glucose conditions. SHH inhibition was less effective under high glucose conditions. SHH treatment increased growth less in cells from hypertension and cardiovascular disease patients. Corpora cavernosa smooth muscle from prostatectomy, diabetic, and Peyronie's patients increased in response to SHH treatment in a similar manner, suggesting that SHH treatment would be beneficial to enhance smooth muscle regeneration in patients with ED of multiple origins. There is a threshold concentration of SHH protein above which smooth muscle growth is enhanced. Lipid modified SHH engendered a more robust growth response. High glucose conditions that may be present in under controlled diabetic patients would not detract from SHH usefulness as a protectant for corpora cavernosal morphology. Understanding how human corpora cavernosal tissue responds to SHH treatment is critical for clinical translation to ED patients. No [ABSTRACT FROM AUTHOR]