Tumour budding, uPA and PAI-1 are associated with aggressive behaviour in colon cancer.

AIMS: The proteases PAI-1 and uPA play a major role in extracellular matrix degradation, which facilitates tumour progression. Tumour budding is a histomorphological expression of enhanced tumour cell migration. MATERIALS AND METHODS: To investigate their prognostic value for and correlation with colon cancer, a prospective study was performed. We analysed tissue levels of uPA and PAI-1 of 55 colon cancer tumours employing a commercially available enzyme-linked immunosorbent assay (ELISA). Tumour budding was analysed on cytokeratin-stained slides. RESULTS: There was a strong correlation between uPA and tumour budding (R = 0.440; P < 0.001). uPA levels were increased in high grade tumours, whereas PAI-1 was elevated in cases with venous invasion (P = 0.004 and P = 0.028). PAI-1 values and tumour budding are associated significantly with the occurrence of distant metastases (P < 0.001 and P = 0.034, respectively). Tumour budding was significantly associated with lymph node metastases (P = 0.034). Multivariate analysis revealed PAI-1 and lymph node metastases to be independently predictive of distant metastases (P = 0.007 and P = 0.004, respectively). CONCLUSIONS: The results of our study show that tumour budding and the plasmin/plasminogen system are related. PAI-1 was independently predictive for the occurrence of distant metastasis.

Cloning and characterization of cDNAs coding for the heavy and light chains of a monoclonal antibody specific for Pseudomonas aeruginosa outer membrane protein I.

A set of seven monoclonal antibodies (MAb) directed against outer membrane proteins of Pseudomonas aeruginosa has been examined by Western blot analysis, indirect immunofluorescence tests and subclass typing. The hybridoma cell line secreting MAb 6A4, which reacts with outer membrane protein I, belongs to the IgG2a subclass and crossreacts with the 17 P. aeruginosa serotypes as listed in the International Antigenic Typing System, was selected as source for the preparation of poly(A)+RNA which in turn was used as template for cDNA synthesis and cloning. Full length cDNA clones of the gamma heavy chain as well as the kappa light chain were obtained and characterized by nucleotide sequence analysis. The complete cDNA sequences coding for the heavy and light chains will be the prerequisite for the construction and heterologous expression of a chimeric human-mouse monoclonal antibody which might be used in therapy of P. aeruginosa infections.

Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients.

Twenty-one high-risk patients with primary stage II/III breast cancer were treated with high-dose chemotherapy comprising etoposide, ifosfamide, carboplatin and epirubicin (VIC-E). Tumor cells of epithelial origin were analyzed using the monoclonal antibodies CK2 (IgG1) and A45-B/B3 (IgG1) against cytokeratin (CK) components in bone marrow (BM) aspirates prior to chemotherapy, and in peripheral blood stem cell transplants (PBSCT). They were separated after the first (21/21 patients) and the second cycle (16/21 patients) of induction chemotherapy with VIP-E (etoposide, ifosfamide, cisplatin, epirubicin). Preliminary results showed CK positive tumor cells in 40% (14/35) of the analyzed transplants. In 7/12 (58.3%) patients, CK positive tumor cells were detectable in BM prior to treatment. Sixteen patients were separated after the 1st and 2nd cycle of VIP-E. PBSCT of 14/16 patients were assessable for presence of CK positive tumor cells. Our preliminary results demonstrate a lower tumor cell contamination of PBSCT separated after the 2nd cycle of induction therapy (14.3%) compared to contamination after the first induction therapy (64.3%). To date, 4/21 patients have experienced a relapse, and three of these patients had tumor cell positive transplants. Due to the small patient number only a trend towards a superior relapse-free survival in the patient group with CK negative transplants can be shown by Kaplan-Meier analysis.

The production of porphyrins from delta-aminolaevulinic acid by Haemophilus parainfluenzae.

Porphyrin production by the haemin-independent Haemophilus parainfluenzae in the diagnostic porphyrin test, which determines the X-factor requirement in Haemophilus spp., was analysed quantitatively by applying modern high-performance liquid chromatographic (HPLC) methods. Ion-pair reversed-phase HPLC enabled the simultaneous separation of all porphyrin intermediates and their isomers of haem biosynthesis produced by the bacteria. The pH-dependence of porphyrin production and the respective composition of the porphyrins within the bacterial cells and in the supernate were investigated. A pH optimum of 6.9-8.0 for the production of porphyrins was found and there were marked differences in the porphyrin profiles at different pH values.

The integration of the transfer of frozen embryos with artificial insemination in a commercial beef herd.

This trial was designed to examine the integration of the transfer of frozen embryos and artificial insemination (A.I.) in a commercial herd of beef cows. Embryos were collected nonsurgically from 15 superovulated cows and all embryos judged viable (n = 69) were frozen in 0.25 ml plastic A.I. straws. Glycerol was used as a cryoprotectant and the embryos were frozen with omission of the traditional "seeding" step. On three consecutive days immediately preceding the onset of a 60-day breeding season, 54 embryos were transferred to multiparous cows with calves at foot. The remaining 82 cows in the same herd were used as controls. Cows receiving embryos were maintained under observation, but not inseminated for 21 days, after which cows exhibiting signs of estrus were inseminated up to Day 60. Cows in the control group were inseminated at observed estrus throughout the 60-day period. The overall pregnancy rate for the experimental and control groups did not differ (77.8 % and 73.2 % respectively). Of the experimental cows, 24.1 % conceived to the embryo transfer, and 53.7 % to A.I. The mean calving to conception interval for the experimental cows was 96.3 days (median 101 days) and for the control cows it was 92.3 days (median 88 days). The calving pattern in the experimental cows was biphasic, calvings resulting from embryo transfer of A.I. accounting for the two peaks. Reconception rates in the following breeding season were not different between the embryo transfer or A.I. groups or between experimental and control herds. Weaning masses of calves born of embryo transfer were significantly higher than the other groups, but calves resulting from A.I. in the experimental herd were not different from calves in the control herd. It was concluded that the transfer of frozen embryos could be successfully integrated into an A.I. program with a limited breeding season without detrimentally affecting the overall reproductive performance of the recipient herd.

[The current status of vaccination and antibiotic prophylaxis in splenectomy. I: Adults]. / Standortbestimmung der Impf- und Antibioticaprophylaxe bei Splenektomie. Teil I: Erwachsene.

The prevention of severe late postsplenectomy infection by vaccination and antibiotic prophylaxis is controversial. This paper reviews the literature and presents a protocol for postsplenectomy adults. Prophylaxis should be adapted to the underlying disease. Every patient has to be informed about the consequences of splenectomy. Depending on the underlying disease, vaccination is recommended against pneumococci, Hemophilus influenzae and Neisseria meningitidis. Long-term antibiotic prophylaxis is not advised in adults.

Causes of variation of copper, iron, manganese, zinc and magnesium levels in bovine livers. 3. The effects of locality.

The effects of locality on the copper, iron, manganese, zinc and magnesium levels in 407 bovine caudate lobe liver samples preserved in formalin for differing storage periods were examined. The mineral determinations, expressed on the wet basis (WB), were made by atomic absorption spectrophotometry after wet ashing of the liver. Two hundred and ten of the liver samples were from cattle from one farm (Farm 1) the remaining 197 cattle being from another farm (Farm 2). The copper, iron and magnesium levels were taken as indicative of the hepatic concentrations at slaughter. Locality had a significant effect (P less than 0,05) on the copper, iron and magnesium levels. All copper levels on Farm 1 fell well below the accepted minimum (33,0 mg/kg). The deficiency appeared to be secondary with the possible implication of sewage effluent. In terms of biological variation the different iron levels appeared of minor importance and no inverse relationship was found between iron and copper. The manganese and zinc levels were interpreted with caution due to the significant differences reported in their hepatic concentrations after six months of storage in formalin. Extremely high zinc levels in individual animals could have been associated with sewage effluent.

The persistence of serological reactions following heifer vaccination with Brucella abortus strain 19 on brucellosis-free farms.

Of 392 heifers vaccinated between 4 and 9.5 months of age and tested at 22 months or older, 3 (0.77%) had titres in the complement fixation test of between 30 and 98 international units per millilitre. The results confirm that persistent serological reactions can be a problem even in brucellosis-free herds. To reduce the incidence of these problem reactors, it is suggested that: in clean herds the age of inoculation be reduced to 3 months of age; the age at first testing be delayed as long as possible, but be done before the heifers are 4 months pregnant, c) consideration be given to reducing the vaccine dose for all heifers from the present 4 to 12 x 10(10) to between 3 x 10(8) and 3 x 10(9) and syringes used for strain 19 vaccination be exclusively reserved for this purpose to prevent the possibility of antigenic residues acting as a booster when used for other vaccinations.

Erythrocyte glutathione peroxidase activity as an indicator of selenium status in an intensively-managed beef herd.

A survey of erythrocyte glutathione peroxidase (GPx) activity was undertaken in a herd of crossbred cattle after 3 cases of white muscle disease had been diagnosed. All animals examined, appeared to be deficient in selenium, relative to control animals sampled. Oral administration of sodium selenite or sodium biselenite was followed within 30 d by an increase in erythrocyte GPx activity. Despite an apparent improvement in the selenium status of cows after supplementation with selenium, no difference was seen in the conception rate after artificial insemination.

Chlamydiosis in a beef herd.

Chlamydial infection in a large beef herd is illustrated and discussed. The pre-natal and post-natal losses that occurred during 1972 are highlighted. Total pre-natal losses for the nine calving herds comprising 2,915 animals varied between 3,7% and 12,4%. Between 1,2% and 11,4% of the calves born alive in the nine herds died before weaning with 70% of the losses occurring within the first three weeks of life. Chlamydial organisms were demonstrated in pre-natal and post-natal losses from all nine herds. Clinical manifestations and lesions involving the intestinal, respiratory, nervous, skeletal, reticulo-endothelial and urinary systems were observed in chlamydia-infected calves. Invariably at autopsy in chlamydia positive cases there was some degree of a fibrinous inflammatory process present. Serological evidence showed that chlamydial organisms had been present on both farms prior to the 1972 investigations.

Brucellosis serology: reduced dose S19 vaccination of yearling heifers versus the use of the standard dose at 5-7 months of age in a clean herd.

In the 427 heifers vaccinated at 12 to 14 months of age with the reduced dose (3 X 10(8) to 1 X 10(9) viable organisms per dose) 3 had antibody titres in the complement fixation test of 196, 688 and 748 IU ml-1, respectively, at 5 months post-inoculation. At 23 to 25 months of age 8/128 (6.3%) of these heifers had rose bengal test (RBT) reactions, at least 6/128 (4.7%) a serum agglutination test (SAT) antibody titre in excess of 30 IU ml-1 and 1/128 had a suspicious reaction in the complement fixation test (CFT) of 30 to 49 IU ml-1. In the 116 heifers inoculated with the standard dose (4 to 12 X 10(10) viable organisms) at 5 to 7 months of age and tested at 22 to 24 months of age, a reactor rate of 37/116 (31.9%) in the RBT was seen. At least 34/116 (29.3%) had antibody titres in excess of 30 IU ml-1 in the SAT. Only 1 animal had a suspicious reaction in the CFT of 18 to 24 IU ml-1. No abortions could be attributed to the vaccine strain and no other isolates of the vaccine strain were made. The possible role of vaccine residues in syringes as a cause of persistent reactions is discussed. The use of disposable syringes is recommended. On serological grounds there appears to be little advantage in using the reduced dose vaccine in yearling heifers as opposed to the standard dose vaccination of 5 to 7 month old heifers.

Chronic splenitis as a possible complicating factor in calves inoculated with an attenuated redwater vaccine.

In two consecutive years a total of 3810 calves were inoculated with the Onderstepoort redwater live-blood vaccine, containing attenuated strains of Babesia bigemina and Babesia bovis. Twenty calves died 8-13 days later due to B. bigemina-infection. Necropsies were performed on 19 of these animals. In 11 calves the spleen was distinctly smaller than normal and its morphology was distorted, and showed changes of chronic splenitis. Histological lesions in the spleens of 6 of the 11 calves are described. The potentiating effect of the splenic lesions to reduce the ability to contain the vaccine reaction, is discussed.

[Impending Coumarin-necrosis in a patient with heterozygous protein C deficiency type I]. / Drohende Cumarinnekrose bei einer 24-jährigen Patientin mit stattgehabter Beinvenenthrombose.

Painful pink or magenta colored skin lesions characterized by a clear line of demarcation between the affected area and surrounding tissue appearing under therapy with coumarins may be a sign for evolving coumarin-necrosis. Immediate treatment with a protein C preparation in patients with protein C deficiency can prevent necrosis.

[Meningitis caused by Streptococcus suis]. / Bakterielle Meningitis durch Streptococcus suis.

HISTORY AND ADMISSION FINDINGS: A 42-year-old man was admitted to hospital because of headache and fever for two days and showing signs of meningism. As a butcher he had had frequent cuts on his hands and forearms. INVESTIGATIONS AND DIAGNOSIS: Analysis of the cerebrospinal fluid revealed an elevated leukocyte count (81% segmented neutrophils). Large numbers of Gram-positive bacteria were found in both cerebrospinal fluid and blood culture, which were subsequently identified as Streptococcus suis (S. suis) by biochemical methods. The hypothesis of bacterial meningitis was thus confirmed. TREATMENT AND CLINICAL COURSE: The patient was treated with ampicillin, initially combined with ceftriaxone. Hearing impairment and vestibular dysfunction - a frequent complication after infection with S. suis - were observed as sequelae, indicating involvement of the vestibulocochlear nerve. CONCLUSION: Infections caused by S. suis are rare in Europe but must be taken into account if an occupational exposition to pigs is established.