Combining the production of L-lactic acid with the production of feed protein concentrates from alfalfa.

The production of L-lactic acid was investigated in combination with the production of protein concentrates in the frame of a green biorefinery for efficient utilization of grasses and legume crops. Alfalfa green juice was the sole substrate utilized for initial lactic acid fermentation with Lactobacillus salivarius, Lactobacillus paracasei or Bacillus coagulans in order to drop the pH and precipitate the plant proteins present in the juice. Afterwards, proteins were separated by microfiltration with 40-42% of protein recovery into protein concentrates, suited for feeding monogastric animals. The (residual) brown juice was investigated as source of nutrients for producing L-lactic acid from glucose or xylose with B. coagulans A107 or B. coagulans A166, respectively. Fermentation of glucose (30, 60, 100 g L-1) resulted in productivities of 2.8-4.0 g L-1 h-1 and yields of 0.85-0.91 g LA per g consumed glucose. Fermentation of xylose (30, 60 g L-1) resulted productivities of 1.1-2.3 g L-1 h-1 and yields of 0.83-0.88 g LA per g consumed xylose. Comparing different brown juices, initial green juice fermentation with B. coagulans is recommended if the brown juice is to be used for producing L-lactic acid. Based on our results, it is possible to combine protein recovery with lactic acid production, and the brown juice proved to be a good nutrient source for L-lactic acid production with high optical purities.

The impact of bariatric surgery on quality of life: a systematic review and meta-analysis.

This study aims to review the obesity literature in order to assess the impact of bariatric surgery on quality of life and the between-study variation by examining the standardized mean magnitude of effect in change in the levels of quality of life. The following databases EMBASE, PubMed, PsycINFO, CINAHL, the Cochrane Library and Web of Science were systematically searched for studies examining change in quality of life in adults receiving bariatric surgery for obesity. Seventy-two studies were included with a total of 9,433 participants treated for obesity with bariatric surgery. The average impact of bariatric surgery on quality of life corresponded to an effect size of 0.88 (95% CI: 0.80-0.96), indicating that bariatric surgery has a significant positive influence on quality of life in general. The impact varied considerably across studies with bariatric surgery showing a significantly greater positive influence on physical quality of life compared to mental quality of life. Bariatric surgery is effective in improving quality of life, especially when looking at physical well-being. Greater focus on the psychological well-being of the person undergoing surgery for obesity may lead to a better post-surgery prognosis for more people.

Fermentation in 24-well plates is an efficient screening platform for filamentous fungi.

UNLABELLED: Fermentation by filamentous fungi in Erlenmeyer flasks is a favoured method for comparing different fermentation conditions. However, significant inter-flask variation often occurs when using Erlenmeyer flasks, which makes the comparison of fermentation product levels less reliable. We have investigated the use of a 24-well plate method for citric acid, ethanol and glycerol batch fermentation using the filamentous fungi Aspergillus carbonarius and compared the relative standard deviation (RSD) from sextuplicates obtained using Erlenmeyer flasks and 24-well plates. The production levels using the Erlenmeyer flasks showed a combined average RSD of 29%, which is two and a half-fold higher than what was measured using the 24-well plates showing an average RSD of 12%. We conclude that fermentation in 24-well plates is a more reliable screening method for metabolite production by filamentous fungi and possibly for screening metabolites in general. SIGNIFICANCE AND IMPACT OF THE STUDY: Fermentation studies with filamentous fungi and especially screening experiments often struggle with high inter-vessel variations in metabolite production. This study compares two different types of frequently used screening methods namely batch fermentation in Erlenmeyer flasks with batch fermentation in 24-well plates. The results demonstrate that the variance potentially can be reduced two and a half-fold using 24-well plates leading to improved resolution when testing the impact of varying fermentation parameters on product formation.

Rhizobacterially induced protection of watermelon against Didymella bryoniae.

AIMS: To identify rhizobacteria from the Mekong Delta of Vietnam, which can systemically protect watermelon against Didymella bryoniae and elucidate the mechanisms involved in the protection conferred by isolate Pseudomonas aeruginosa 23(1-1). METHODS AND RESULTS: Bacteria were isolated from watermelon roots and their antagonistic ability tested in vitro. Of 190 strains, 68 were able to inhibit D. bryoniae by production of antibiotics. Four strains were able to reduce foliar infection by D. bryoniae when applied to watermelon seeds before sowing. Strain Ps. aeruginosa 23(1-1) was chosen for investigations of the mechanisms involved in protection and ability to control disease under field conditions. In the field, the bacterium was able to significantly reduce disease in two consecutive seasons and increase yield. Furthermore, it colonized watermelon plants endophytically, with higher numbers in plants infected by D. bryoniae than in noninoculated plants. To elucidate the mechanisms involved in protection, the infection biology of the pathogen was studied in bacterially treated and control plants. Pseudomonas aeruginosa 23(1-1) treatment inhibited pathogen penetration and this was associated with hydrogen peroxide accumulation, increased peroxidase activity and occurrence of new peroxidase isoforms, thus indicating that resistance was induced. CONCLUSIONS: The endophytic bacterium Ps. aeruginosa 23(1-1) can control D. bryoniae in watermelon by antibiosis and induced resistance under greenhouse and field conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that rhizobacteria from native soils in Vietnam can be used to control gummy stem blight of watermelon through various mechanisms including induction of resistance.

First Report of Anthracnose Fruit Rot Caused by Colletotrichum acutatum on Strawberry in Denmark.

Strawberry (Fragaria × ananassa) is the most important small fruit crop in Denmark. The quarantine pathogen Colletotrichum acutatum was detected for the first time in June 2000 in Denmark in a production field on the island of Falster. Strawberry plants of cv. Kimberly showed typical symptoms of anthracnose fruit rot. On mature fruits, brown-to-black lesions with spore masses that were orange to salmon in color were observed. Mummified berries were also observed. The fungus was isolated and identified on the basis of morphological characteristics, and identification was confirmed using enzyme-linked immunosorbent assay at the Central Science Laboratory, York, U.K. Species-specific polymerase chain reaction with the C. acutatum-specific primer pairs acut1/col2 (1) and CaInt2/ITS4 (3) also supported the identification. Additionally, the internal transcribed spacer regions, ITS1 and ITS2, of the ribosomal DNA were sequenced in both directions (GenBank Accession No. AY818361). Homology searches with this sequence using BLAST also confirmed the identity. Colonies grown on potato dextrose agar developed white-to-grey aerial mycelium with salmon-colored spore masses, and were beige to black on the reverse side. Conidia were 11.3 (7.3 to 16.6) µm × 3.9 (2.5 to 5.2) µm, hyaline, cylindrical with at least one pointed end, and aseptate. Mycelial growth rate was 8.4 mm per day at 25°C which is similar to earlier reports (2). Spray-inoculated (106 conidia per ml) strawberry fruits cv. Elsanta developed brown, sunken, irregular lesions with salmon-colored acervuli after 2 to 5 days at 25°C. Koch's postulates were fulfilled since the reisolated fungus from these lesions developed the same morphological characteristics as described above. To our knowledge, this is the first report of C. acutatum in Denmark. References: (1) P. V. Martinez-Culebras et al. J. Phytopathol. 151:135, 2003. (2) B. J. Smith et al. Plant Dis. 74:69, 1990. (3) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996.

Prediction of left ventricular functional recovery by dobutamine echocardiography, F-18 deoxyglucose or 99mTc sestamibi nuclear imaging in patients with chronic myocardial infarction.

BACKGROUND: Currently, several modalities are available to predict viability, however, studies comparing various modalities validated by functional recovery after revascularization are scarce. This study analyzed the relative merits of low-dose dobutamine echocardiography, F-18 deoxyglucose (FDG) positron emission tomography (PET) and (99m)Tc sestamibi single-photon emission computed tomography to predict functional recovery after revascularization in patients with chronic myocardial infarction. METHODS: Patients with chronic coronary occlusion (duration: 3.1 +/- 4.8 years) and impaired left ventricular function (ejection fraction: 42 +/- 13%) underwent low-dose dobutamine echocardiography (20 microg/kg/min), FDG-PET and (99m)Tc sestamibi imaging before revascularization. Revascularization was performed irrespective of any viability data. Follow-up angiography was obtained 4.8 +/- 2.5 months after revascularization. RESULTS: Viability analysis was performed in 34 patients with patent target vessel at follow-up, of whom 9 (27%) exhibited functional recovery on left ventricular angiography. For dobutamine echocardiography, improvement of >/=2 adjacent akinetic segments resulted in improved sensitivity of 89% and specificity of 80% to predict functional recovery. For glucose metabolism, FDG uptake >55% was an optimal threshold yielding a sensitivity of 89% and a specificity of 68%. With respect to perfusion, (99m)Tc sestamibi uptake >60% was the best cutoff resulting in a sensitivity and a specificity of 56 and 88%, respectively. A concordant match of FDG >55% and of (99m)Tc sestamibi >50% resulted in optimized sensitivity (78%) and specificity (80%) with dual imaging. CONCLUSIONS: Recovery of chronically dysfunctional myocardium can be predicted with high accuracy by stimulation of contractile reserve or by concordant match of preserved glucose metabolism and residual perfusion.

UP-PCR cross blot hybridization as a tool for identification of anastomosis groups in the Rhizoctonia solani complex.

A universally primed (UP)-PCR cross hybridization assay was developed for rapid identification of isolates of Rhizoctonia solani into the correct anastomosis group (AG). Twenty-one AG tester isolates belonging to 11 AGs of R. solani were amplified with a single UP primer which generated multiple PCR fragments for each isolate. The amplified products were spotted onto a filter, immobilized and used for cross hybridization against amplification products from the different isolates. Isolates within AG subgroups cross hybridize strongly, whereas between different AGs little or no cross hybridization occurs. Sixteen Rhizoctonia isolates from diseased sugar beets and potatoes were identified using the assay. The results were supported by restriction fragment length polymorphism analysis of the ITS1-5.8S-ITS2 region of the nuclear encoded ribosomal DNA. Through standardization and use of quick non-radioactive labeling techniques, the UP-PCR cross hybridization assay has potential for routine use by modern DNA chip technology.

Myocardial viability assessed by positron emission tomography in infants and children after the arterial switch operation and suspected infarction.

OBJECTIVES: The aim of the study was to assess regional glucose metabolism and contractile function by gated positron emission tomography using fluoro-18-deoxyglucose (FDG-PET) in pediatric patients after the arterial switch operation and suspected myocardial infarction. BACKGROUND: Morbidity and mortality after the arterial switch operation for transposition of the great arteries are often related to impaired coronary function. Justification of high-risk revascularization procedure in infancy requires thorough evaluation of myocardial viability. Although PET is state-of-the-art for evaluation of myocardial viability in adults there are no reports on its impact and feasibility in infants and children. METHODS: We applied electrocardiogram-triggered FDG-PET for assessment of metabolic and functional status of the myocardium in seven infants and seven children. Glucose metabolism, wall motion and wall thickening were evaluated visually and quantitatively on the basis of parametric 3-D images. Additionally, single-photon emission computed tomography perfusion scan was performed in six children. RESULTS: In two of seven infants, FDG-PET demonstrated viable myocardium in akinetic or hypokinetic regions corresponding to a coronary artery stenosis or occlusion. Therefore, indication for revascularization was derived from this finding. In six of the seven children, impaired glucose uptake reflecting myocardial scarring was present. Two patients had pathological findings on coronary angiography and signs of ischemia but were not suitable for revascularization. CONCLUSIONS: Myocardial viability and contractile function can be assessed simultaneously by gated FDG-PET even in infant hearts. This method contributes pertinent information to guide further therapy after the arterial switch operation and suspected myocardial infarction.

Identification of a universally primed-PCR-derived sequence-characterized amplified region marker for an antagonistic strain of Clonostachys rosea and development of a strain-specific PCR detection assay.

We developed a PCR detection method that selectively recognizes a single biological control agent and demonstrated that universally primed PCR (UP-PCR) can identify strain-specific markers. Antagonistic strains of Clonostachys rosea (syn. Gliocladium roseum) were screened by UP-PCR, and a strain-specific marker was identified for strain GR5. No significant sequence homology was found between this marker and any other sequences in the databases. Southern blot analysis of the PCR product revealed that the marker represented a single-copy sequence specific for strain GR5. The marker was converted into a sequence-characterized amplified region (SCAR), and a specific PCR primer pair was designed. Eighty-two strains, isolated primarily from Danish soils, and 31 soil samples, originating from different localities, were tested, and this specificity was confirmed. Two strains responded to the SCAR primers under suboptimal PCR conditions, and the amplified sequences from these strains were similar, but not identical, to the GR5 marker. Soil assays in which total DNA was extracted from GR5-infested and noninoculated field soils showed that the SCAR primers could detect GR5 in a pool of mixed DNA and that no other soil microorganisms present contained sequences amplified by the primers. The assay developed will be useful for monitoring biological control agents released into natural field soil.

Identification of Trichoderma strains from building materials by ITS1 ribotyping, UP-PCR fingerprinting and UP-PCR cross hybridization.

To study the role of Trichoderma in sick building syndrome, it is essential to be able to accurately identify species. Forty-four strains of Trichoderma spp. isolated from Danish buildings damaged by water leaks were identified using ITS1 ribotyping and universally primed PCR, UP-PCR. Ribotyping allowed the assignment of the strains into three distinct groups. High similarity of UP-PCR banding profiles of the strains allowed species designation for almost all strains (43 out of 44) when compared with the UP-PCR banding profiles obtained from reference strains of T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum and T. viride. However, cross hybridization of UP-PCR products showed that the latter strain had high DNA homology to the ex-type strain of T. hamatum. The combined approach is a convenient way for reliable identification of Trichoderma strains.

How important is the route of reconstruction after esophagectomy: a prospective randomized study.

OBJECTIVE: A prospective randomized trial was performed to compare retrosternal and posterior mediastinal gastric tube reconstruction with regard to postoperative function and quality of life. METHODS: Twenty-six patients were randomly allocated to either retrosternal (n = 14) or posterior mediastinal (n = 12) reconstruction after gastric tube formation. Radionuclide transit studies were applied to obtain objective functional data and a standardized quality-of-life assessment was performed. RESULTS: Retrosternal reconstruction showed an increased morbidity (15 vs 13 major complications) and mortality (14.2 vs 8.3%). Radionuclide clearance in the supine position was delayed in the gastric tube in general, compared with normal controls (retention index > 40% vs < 10%). There was a significantly higher retention (p < 0.005) in the retrosternal group in the middle third of the tube and the whole tube after intake of the liquid tracer. The retention of the first solid tracer was also higher in the retrosternal group in the middle third of the tube (p = n.s.) and was significantly higher in the whole tube after 30 (p < 0.05) and 60 (p < 0.01) s. This had no significant impact on the patients' quality of life. CONCLUSIONS: The posterior mediastinal route of reconstruction is recommended but curative resection (R0) is mandatory to avoid possible complications due to local tumor relapse. After incomplete resection (R1 or R2) we recommend retrosternal reconstruction for better palliation.

Enhanced immunogenicity of hepatitis B surface antigen by insertion of a helper T cell epitope from tetanus toxoid.

The currently marketed hepatitis B vaccines in the U.S. are based on the recombinant major hepatitis B surface antigen (HBsAg) of hepatitis B virus. Although a large majority of individuals develop protective immunity to HBV-induced disease after three immunizations, routinely a small but a significant percentage of the human population does not respond well to these vaccines. In this report, we describe the generation of a novel HBsAg molecule containing a Th epitope derived from tetanus toxoid (TT). Using recombinant DNA technology. the TT Th epitope (TTe) was inserted into the HBsAg coding sequence. Using a recombinant adenovirus expression system, HBsAg TTe chimeric protein was produced in A549 cells and found to be secreted into culture medium as 22 nm particles. The chimeric HBsAg particles were readily purified by immunoaffinity chromatography and their immunogenicity was evaluated relative to native HBsAg produced in an adenovirus expression system. When evaluated in inbred and outbred strains of mice, HBsAg TTe was shown to enhance several-fold the anti-HBs response relative to native HBsAg. Further enhanced responses were observed in mice primed with TT. This highly immunogenic form of HBsAg has promise as an improved HBsAg subunit vaccine.

Histological verification of positive fluorine-18 fluorodeoxyglucose findings in patients with differentiated thyroid cancer.

Fluorine-18 fluorodeoxyglucose and positron emission tomography (FDG-PET) is a new imaging modality used in the follow-up of patients with differentiated thyroid cancer if the results of (131)I scintigraphy are negative in spite of an elevated thyroglobulin level. The aim of this retrospective analysis was to estimate the value of FDG-PET regarding the operability of patients with positive findings. From January 1994 to October 1997, we investigated 60 patients with differentiated thyroid carcinoma by FDG-PET. Thirteen patients were operated on after positive findings. Most of these lesions were suspected of having lymph-node involvement or local recurrences in the thyroid bed. One patient showed a solitary distant metastasis in the scapula. Thirteen of 16 operations in these 13 patients confirmed the suspected involvement of thyroid cancer. The false-positive findings were caused by inflamed lymph nodes in two cases and benign thymus tissue in one case. We conclude that PET is a useful diagnostic tool to guide early surgical therapy in patients with (131)I negative differentiated thyroid carcinoma.

[Computerized tomography and F-18-FDG positron emission tomography in staging of malignant lymphomas: a comparison]. / Computertomographie und F-18-FDG-Positronen-Emissions-Tomographie im Staging maligner Lymphome: ein Vergleich.

PURPOSE: To determine the value of F-18-FDG-positron emission tomography (FDG-PET) compared with computed tomography (CT) in the staging of malignant lymphomas. MATERIAL AND METHOD: 50 patients with biopsy-proven lymphoma were studied with FDG-PET and CT. The results in initial, posttherapeutic and staging of recurrence were compared. RESULTS: 37 of 65 FDG-PET were identical with CT. 28 studies showed differences. 14 post-therapeutically and one of the initial studies led to downstaging by FDG-PET were as upstaging resulted in one case of initial staging. In two cases false positive pulmonary FDG accumulations caused an upstaging. CONCLUSION: FDG-PET was at least comparable to CT in recording the extension of a newly diagnosed lymphoma, or its recurrence. Upstaging according to FDG-PET occurred only once in initial staging. FDG-PET plays its most important role in the evaluation of residual mass in CT after therapy by accumulating FDG in viable tumour rather than in fibrotic tissue. 14 cases of downstaging according to FDG-PET resulted.

Heat shock transiently enhances the synthesis rate of Sis1p, a ribosome-associated DnaJ protein in the oleagenous yeast Apiotrichum curvatum.

DnaJ proteins have been localized in different intracellular compartments of eukaryotes. In Apiotrichum curvatum, a fat-storing yeast, we found a DnaJ homolog associated with ribosomes and large cytosolic complexes as well. Using a plant DnaJ probe and a cDNA library constructed from poly(A)(+)-RNA of A. curvatum grown on oleate we isolated a SIS1 cDNA coding for a 39.5 kDa protein. The putative protein contains neither a zinc finger motif nor a CAAX motif but is characterized by a J-domain at the N-terminal region and a large G-rich region in the middle part of the molecule. Heat shock applied for 1 h resulted in a pronounced but transient increase of the SIS1 mRNA. An antiserum was raised against the bacterially expressed protein. Cell fractions from A. curvatum were further separated by sedimentation centrifugation on sucrose gradients. Analysing the sub-fractions, we detected Sis1p mainly associated with ribosomes, and with particles sedimenting at approximately 200S. Hsp70 was found to be associated with the 200S fraction. The respective cytosolic A. curvatum Hsp70 cDNA was cloned and sequenced. High salt conditions caused the removal of Hsp70 and Sis1p from the 200S complexes. Mild RNase treatment of the 200S fraction afforded monosomes and 200S complexes unaffected by RNase. Heat shock led to a pronounced increase in the rate of de novo synthesis. However, due to the large pools of Sis1p on ribosomes and large cytosolic complexes, the increase in gene activation did not lead to a significant change of the total amount of Sis1p.

Induction of neutralizing antibodies to T-cell line-adapted and primary human immunodeficiency virus type 1 isolates with a prime-boost vaccine regimen in chimpanzees.

Five chimpanzees were immunized by administration of one or more intranasal priming doses of one to three recombinant adenoviruses containing a gp160 insert from human immunodeficiency virus type 1 (HIV-1) MN (HIV-1MN) followed by one or more boosts of recombinant HIV-1SF2 gp120 delivered intramuscularly with MF59 adjuvant. This regimen resulted in humoral immune responses in three of five animals. Humoral responses included immunochemically active anti-H1V-1 antibodies (Abs) directed to recombinant gp120 and neutralizing Abs reactive with T-cell-line-adapted HIV-1MN and HIV-1SF2. In addition, neutralizing activity was detected to the two homologous primary isolates and to two of three heterologous primary isolates which, like the immunizing strains, can use CXCR4 as a coreceptor for infection. The three animals with detectable neutralizing Abs and a fourth exhibiting the best cytotoxic T-lymphocyte response were protected from a low-dose intravenous challenge with a cell-free HIV-1SF2 primary isolate administered 4 weeks after the last boost. Animals were rested for 46 weeks and then rechallenged, without a boost, with an eightfold-higher challenge dose of HIV-1SF2. The three animals with persistent neutralizing Abs were again protected. These data show that a strong, long-lived protective Ab response can be induced with a prime-boost regimen in chimpanzees. The data suggest that in chimpanzees, the presence of neutralizing Abs correlates with protection for animals challenged intravenously with a high dose of a homologous strain of HIV-1, and they demonstrate for the first time the induction of neutralizing Abs to homologous and heterologous primary isolates.

An adenovirus-simian immunodeficiency virus env vaccine elicits humoral, cellular, and mucosal immune responses in rhesus macaques and decreases viral burden following vaginal challenge.

Six female rhesus macaques were immunized orally and intranasally at 0 weeks and intratracheally at 12 weeks with an adenovirus type 5 host range mutant (Ad5hr)-simian immunodeficiency virus SIVsm env recombinant and at 24 and 36 weeks with native SIVmac251 gp120 in Syntex adjuvant. Four macaques received the Ad5hr vector and adjuvant alone; two additional controls were naive. In vivo replication of the Ad5hr wild-type and recombinant vectors occurred with detection of Ad5 DNA in stool samples and/or nasal secretions in all macaques and increases in Ad5 neutralizing antibody in 9 of 10 macaques following Ad administrations. SIV-specific neutralizing antibodies appeared after the second recombinant immunization and rose to titers > 10,000 following the second subunit boost. Immunoglobulin G (IgG) and IgA antibodies able to bind gp120 developed in nasal and rectal secretions, and SIV-specific IgGs were also observed in vaginal secretions and saliva. T-cell proliferative responses to SIV gp140 and T-helper epitopes were sporadically detected in all immunized macaques. Following vaginal challenge with SIVmac251, transient or persistent infection resulted in both immunized and control monkeys. The mean viral burden in persistently infected immunized macaques was significantly decreased in the primary infection period compared to that of control macaques. These results establish in vivo use of the Ad5hr vector, which overcomes the host range restriction of human Ads for rhesus macaques, thereby providing a new model for evaluation of Ad-based vaccines. In addition, they show that a vaccine regimen using the Ad5hr-SIV env recombinant and gp120 subunit induces strong humoral, cellular, and mucosal immunity in rhesus macaques. The reduced viral burden achieved solely with an env-based vaccine supports further development of Ad-based vaccines comprising additional viral components for immune therapy and AIDS vaccine development.

Long-term protection of chimpanzees against high-dose HIV-1 challenge induced by immunization.

A combination AIDS vaccine approach consisting of priming with adenovirus-HIV-1MN gp160 recombinants followed by boosting with HIV-1SF2 gp120 was evaluated in chimpanzees. Long-lasting protection, requiring only three immunizations, was achieved against a low-dose challenge with the SF2 strain of HIV-1 and a subsequent high-dose SF2 challenge administered 1 year later without an intervening boost. Notably, neutralizing antibody responses against both clinical and laboratory isolates developed in three chimpanzees and persisted until the time of high-dose challenge. The possibility that cytotoxic T-lymphocytes contribute to low-dose protection of a chimpanzee lacking neutralizing antibodies is suggested. Our results validate the live vector priming/subunit booster approach and should stimulate interest in assessing this combination vaccine approach in humans.

[Transmyocardial laser revascularization--a treatment option for coronary heart disease?]. / Transmyokardiale Laserrevaskularisation--Behandlungsoption der koronaren Herzerkrankung?

Transmyocardial laser revascularization (TMR) is a new therapeutic principle for patients with coronary artery disease and no possibility of conventional revascularization with CABG or PTCA. The clinical value of the method is not known. Therefore we investigated all 46 patients treated with sole TMR in our center using clinical investigation, LV and coronary angiography, right heart catheterization, MIBI perfusion imaging and myocardial FDG-PET pre- and 6 months post TMR. 117 patients judged not suitable for conventional revascularization procedures were submitted for TMR. The indication for the procedure was reevaluated in every case. 52 patients (mean EF 41 +/- 16%) could be further treated by intensified anti-anginal medication, seven patients received bypass grafts, four patients had PTCA, three patients were listed for heart transplantation, and five patients had a combined CABG plus TMR. Only 46 (38% of the submitted patients, mean EF 55 +/- 15%) were accepted for sole TMR. CCS class of these patients was 3.3 +/- 0.4, mean age was 63.6 +/- 7.3 years, 70% were males. The postoperative mortality within 30 days was 5/46 (10.8%); 9/46 patients (19.5%) suffered from perioperative myocardial infarction. Other complications were ventricular fibrillation in two cases on the second postoperative day and a rupture of the spleen on the 14th postoperative day. 8/46 patients (17%) had wound infections. Survivors showed an improvement in their CCS class (1.9, 2.1, 1.9 after 3, 6 and 12 months, respectively, mean observation time 0.61 +/- 0.4 years). These patients were able to perform bicycle stress tests significantly longer (98 s +/- 9 pre versus 120 +/- 13 s post TMR, p = 0.01). Angiographic EF fell from 57.8% +/- 15% to 52.6% +/- 19% (p = 0.02) and the number of hypokinetic chords rose from 23.6 +/- 20.9% to 30.6 +/- 24.1% per patient (p = 0.008), predominantly in the inferior wall. Nuclear studies showed reduced myocardial perfusion and vitality after TMR. Four patients in the TMR group had reintervention (PTCA) because of progression of coronary sclerosis of native vessels. One patient had mitral valve replacement due to severe regurgitation. Kaplan-Meier analysis showed no significant difference in survival between the TMR and the medical group when stratified according to initial ejection fraction. Sudden death and congestive heart failure are the most important causes of mortality. Our data show that TMR improves symptoms and exercise performance of otherwise not treatable patients with diffuse coronary artery disease. Due to a lack of an improvement of cardiac perfusion, function or prognosis TMR should be used only in highly selected cases when conventional methods fail to improve patients symptoms.

Replication and immunogenicity of Ad7-, Ad4-, and Ad5-hepatitis B virus surface antigen recombinants, with or without a portion of E3 region, in chimpanzees.

Human adenovirus vectors containing intact or largely deleted E3 region were used to construct adenovirus-hepatitis B recombinant viruses (Ad-HepB) and shown to produce substantial amount of recombinant protein, hepatitis B surface antigen (HBsAg), in tissue culture. Previously we showed that these viruses were able to elicit good anti-HBs antibodies in a dog model. In the present study, the Ad-HepB viruses were evaluated for replication and immunogenicity in chimpanzees which sustain permissive infection by human adenoviruses. Recombinants containing entire E3 region showed better replication pattern than their E3 deleted counterparts as evidenced by longer duration and high titers of virus shedding. The effect of E3 region was also seen in the antibody titers against HBsAg in that the E3 containing viruses showed better response than the E3 deleted viruses. The importance of E3 region for the development of adenovirus vectored vaccines is further discussed.