In vivo characterisation of two Australian isolates of Marek's disease virus including pathology, viral load and neuropathotyping based on clinical signs.

OBJECTIVE: To evaluate the pathogenicity of Australian Marek's disease virus (MDV) isolate MPF23 (1985) against the reference strain MPF57 based on pathology, viral load and neuropathotyping on the basis of clinical signs. PROCEDURE: Two MDV challenge isolates (MPF57 or MPF23) were administered to unvaccinated specific-pathogen free (SPF) layer chicks on day 5 after hatch at three challenge doses (500, 2000 or 8000 plaque-forming units (pfu)/chick). Mortality, body weight, immune organ weights, MDV load in peripheral blood lymphocytes (PBL) and clinical signs were measured to 56 days post challenge (dpc). RESULTS: MPF23 was the more pathogenic of the two viruses, inducing higher mortality (81% vs 62%) and incidence of MD lesions (100% vs 76%). MPF23 induced earlier, more sustained and more severe neurological signs in the period 26-56 dpc. However, there were few differences during the 0-23 dpc used in the neuropathotyping classification under test. The observed pattern during this earlier period classified both viruses as neuropathotype B, consistent with a very virulent pathotype. MDV load in PBL at 7 and 44 dpc did not differ between virus isolates, but the load at 7 dpc was significantly and negatively associated with time to euthanasia or death. CONCLUSION: MPF23 appears to be as, or more, virulent than the MDV strains isolated over the subsequent two decades. The neuropathotyping system developed in the USA did not clearly differentiate between the two isolates under test; however, extension of the period of assessment of clinical signs beyond 26 dpc did reveal clear differences.

Flaxseed oil supplementation alters the expression of inflammatory-related genes in dogs.

Long chain n-3 fatty acids are beneficial to mammals because of their anti-inflammatory role. However, whether flaxseed oil, which is rich in short chain n-3 fatty acids, has such a role, it has not been extensively examined. This study investigated the supplementation of flaxseed oil on the regulation of genes involved in inflammatory responses such as heat shock proteins (HSP90 and HSP70) and interleukin (IL1ß) in the white blood cells of dogs. Five beagles and 5 greyhounds were supplemented with Melrose(®) flaxseed oil at the rate of 100 mL/kg food for 21 days. The blood was collected at day 0, 15, and 22 following supplementation. The expression of 3 genes was quantified using real-time polymerase chain reaction. Plasma concentrations of fatty acids such as alpha linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid, and arachidonic acid were measured, and their correlations with changes in gene expression were determined. Flaxseed oil supplementation downregulated the expression of HSP90 and IL1ßin greyhounds but showed no significant effect on these genes in beagles. HSP70 remained unchanged in both breeds following the supplementation. Correlations of HSP90 and IL1ßexpression levels with the plasma fatty acid concentrations on day 22 showed a significant negative correlation in greyhounds. Dietary flaxseed oil altered the expression of genes involved in inflammation in white blood cells. Because the expression of the genes may vary in different breeds, it will be useful to consider breed responses to dietary manipulation in canine nutrition management.

Global gene expression profiling reveals genes expressed differentially in cattle with high and low residual feed intake.

Feed efficiency is an economically important trait in beef production. It can be measured as residual feed intake. This is the difference between actual feed intake recorded over a test period and the expected feed intake of an animal based on its size and growth rate. DNA-based marker-assisted selection would help beef breeders to accelerate genetic improvement for feed efficiency by reducing the generation interval and would obviate the high cost of measuring residual feed intake. Although numbers of quantitative trait loci and candidate genes have been identified with the advance of molecular genetics, our understanding of the physiological mechanisms and the nature of genes underlying residual feed intake is limited. The aim of the study was to use global gene expression profiling by microarray to identify genes that are differentially expressed in cattle, using lines genetically selected for low and high residual feed intake, and to uncover candidate genes for residual feed intake. A long-oligo microarray with 24 000 probes was used to profile the liver transcriptome of 44 cattle selected for high or low residual feed intake. One hundred and sixty-one unique genes were identified as being differentially expressed between animals with high and low residual feed intake. These genes were involved in seven gene networks affecting cellular growth and proliferation, cellular assembly and organization, cell signalling, drug metabolism, protein synthesis, lipid metabolism, and carbohydrate metabolism. Analysis of functional data using a transcriptional approach allows a better understanding of the underlying biological processes involved in residual feed intake and also allows the identification of candidate genes for marker-assisted selection.

Oxalate nephropathy in a laboratory colony of common marmoset monkeys (Callithrix jacchus) following the ingestion of Eucalyptus viminalis.

Seven common marmoset monkeys (Callithrix jacchus) from a laboratory colony of 17 died over a period of eight months. Death of six of these monkeys was attributed to kidney failure from an oxalate-induced nephropathy. The epidemiology of this outbreak suggested an exogenous source and there was strong evidence that the source was bark and leaves from an Eucalyptus viminalis tree. Branches of this tree were introduced one month before the first death. The branches were removed one month after deaths commenced, but deaths continued for another five months. Urinalysis of all surviving marmosets at 80 and 122 days after initial contact with the E viminalis branches suggested that these monkeys had renal impairment. In the cases described here, the eating behaviour of common marmosets apparently exposed the animals to toxic levels of oxalate in the bark and leaves of an E viminalis tree.

Effects of the absence of protozoa from birth or from weaning on the growth and methane production of lambs.

Merino ewes (n 108) joined to a single sire were allocated into three flocks, with ewes in one flock being chemically defaunated in the second month of gestation. Single lambs born to defaunated ewes (BF lambs) were heavier at birth and at weaning than lambs born to faunated ewes (F lambs). After weaning, all BF and F lambs were individually housed then half of the F lambs were chemically defaunated (DF lambs). In trial 1, BF, DF and F lambs were offered a concentrate-based diet containing either 14 or 19 % protein for a 10-week period. Wool growth rate of BF lambs was 10 % higher than that of DF or F lambs and was increased 9 % by the high-protein diet. While there was no main effect of protozoa treatment on enteric methane production, there was an interaction between protozoa treatment and diet for methane production. BF and DF lambs produced more methane than F lambs when fed the low-protein diet but when fed the high-protein diet, emissions were less than (BF lambs) or not different from (DF lambs) emissions from F lambs. In trial 2, lambs were offered 800 g roughage per d and, again, methane production was not affected by the presence of protozoa in the rumen. The data indicate that while lambs without rumen protozoa have greater protein availability than do faunated ruminants, there is no main effect of rumen protozoa on enteric methane production by lambs fed either a concentrate or roughage diet.

Salmonella and on-farm risk factors in healthy slaughter-age cattle and sheep in eastern Australia.

OBJECTIVE: To examine healthy slaughter-age cattle and sheep on-farm for the excretion of Salmonella serovars in faeces and to identify possible risk factors using a questionnaire. PROCEDURE: The study involved 215 herds and flocks in the four eastern states of Australia, 56 with prior history of salmonellosis. Production systems examined included pasture beef cattle, feedlot beef cattle, dairy cattle, prime lambs and mutton sheep and animals were all at slaughter age. From each herd or flock, 25 animals were sampled and the samples pooled for Salmonella culture. All Salmonella isolated were serotyped and any Salmonella Typhimurium isolates were phage typed. Questionnaires on each production system, prepared in Epi Info 6.04, were designed to identify risk factors associated with Salmonella spp excretion, with separate questionnaires designed for each production system. RESULTS: Salmonellae were identified in all production systems and were more commonly isolated from dairies and beef feedlots than other systems. Statistical analysis revealed that dairy cattle were significantly more likely to shed Salmonella in faeces than pasture beef cattle, mutton sheep and prime lambs (P<0.05). A wide diversity of Salmonella serovars, all of which have been isolated from humans in Australia, was identified in both cattle and sheep. Analysis of the questionnaires showed access to new arrivals was a significant risk factor for Salmonella excretion on dairy properties. For beef feedlots, the presence of large numbers of flies in the feedlot pens or around stored manure were significant risk factors for Salmonella excretion. CONCLUSION: Dairy cattle pose the highest risk of all the slaughter-age animals tested. Some of the identified risk factors can be overcome by improved management practices, especially in relation to hygiene.

Salmonella Typhimurium persistence in a Hunter Valley dairy herd.

OBJECTIVE: An epidemiological study was undertaken at a Hunter Valley dairy with persistent Salmonella Typhimurium infection. The aim of the study was to identify cattle currently or previously infected with Salmonella, possible sources of the organism, patterns of spread, and husbandry practices that could be improved. METHODOLOGY: Faecal samples, feed, water and environmental samples were cultured for Salmonella and blood samples were tested for antibodies against Salmonella (Dublin and Typhimurium). A questionnaire was designed to identify possible risk factors associated with Salmonella excretion. RESULTS: S Typhimurium was apparently introduced from an old to a new dairy through manure spread as fertiliser. Salmonella apparently persisted in the effluent pond, and the following year clinical cases occurred after pasture, irrigated with water from the pond, was grazed by dry cows, and adult cattle became clinically ill with salmonellosis. The disease spread to other cows and calves. Poor design of calf pens assisted spread of Salmonella from sick to healthy calves. In addition, there was suspected transmission to the dairy farmer's 9-month-old daughter. Salmonellosis on a farm is a potential zoonotic risk to farm workers and their families. There is also the risk that cull cows may carry Salmonella to the abattoir and subsequently into the human food chain. Methods of waste management, and the design of calf pens, were identified as major risk factors that could be improved to minimise the spread of salmonellosis on this property.

Characterization of mesophilic bacilli in faeces of feedlot cattle.

AIMS: To determine the identity and composition of mesophilic Bacillus spp. in faeces sampled from feedlot cattle. METHODS AND RESULTS: Faecal samples from 10 feedlot cattle were analysed. The total aerobic spore count increased from 4.6 x 10(4) CFU g(-1) (before feedlotting, day 0) to 1.6 x 10(6) CFU g(-1) (feedlot for day 76). A total of 150 randomly selected spore isolates (60 each from days 0 and 76 cattle, 30 from feed) were speciated using a Bacillus group-specific PCR-amplified ribosomal DNA restriction analysis technique (Wu et al. 2006). At day 0, Bacillus subtilis and Bacillus cereus predominated with a prevalence of 58.3% and 26.7%, respectively, whereas three species, B. subtilis (50.0%), Bacillus licheniformis (27.6%) and Bacillus clausii (20.0%) predominated in day 76 faecal samples. Of these, only the first two species were present in feed samples at a frequency of 70% and 30% respectively. All B. cereus isolates on day 0, possessed at least one of three enterotoxin genes (nheA, nheB and nheC) but these were completely eliminated after a period of feedlotting. All isolates of B. licheniformis were genotypically heterogeneous according to pulsed-field gel electrophoresis analysis. CONCLUSIONS: Cattle faeces contain large numbers of Bacillus spores representing different mesophilic species. Stable faecal populations of particular Bacillus spp. mimicking those found in feed, were subsequently established by feedlotting. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained and methods used in this study will help to investigate the indigenous Bacillus composition in the gastrointestinal tract of cattle and will further guide the administration of Bacillus probiotics.

A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing Escherichia coli.

AIMS: To investigate interactions between rumen protozoa and Shiga toxin-producing Escherichia coli (STEC) and to ascertain whether it is likely that rumen protozoa act as ruminant hosts for STEC. METHODS AND RESULTS: The presence of stx genes in different microbial fractions recovered from cattle and sheep rumen contents and faeces was examined using PCR. In animals shedding faecal STEC, stx genes were not detected in the rumen bacterial or rumen protozoal fractions. Direct interactions between ruminal protozoa and STEC were investigated by in vitro co-incubation. Rumen protozoa did not appear to ingest STEC, a STEC lysogen or non-STEC E. coli populations when co-incubated. CONCLUSIONS: The ruminal environment is unlikely to be a preferred habitat for STEC. Bacterial grazing by rumen protozoa appears to have little, if any, effect on STEC populations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that ruminal protozoa are unlikely to be a major factor in the survival of STEC in ruminants. They appear as neither a host that protects STEC from the ruminal environment nor a predator that might reduce STEC numbers.

Predictive value of the tumor oxygenation by means of pO2 histography in patients with advanced head and neck cancer.

PURPOSE: The purpose of this analysis was to evaluate the potential of the tumor oxygenation by means of Eppendorf pO2 histography as a predictive test to select patients for treatment alternatives. PATIENTS AND METHODS: Pretreatment tumor pO2 histographies of locoregional lymph node metastases were assessed in 194 patients with squamous cell carcinoma of the head and neck. Included in the analysis were 134 patients who received a primary radio- or radiochemotherapy with a radiation dose of > or = 60 Gy, and who had no distant metastasis at beginning of the therapy. RESULTS: The Cox regression analysis revealed the fraction of pO2 values < or = 2.5 mm Hg (p = 0.004), age (p = 0.04) and radiotherapy/radiochemotherapy (p = 0.03) as significant independent prognostic factors for the survival. The positive and negative predictive values were calculated using different cut-off values of the fraction of pO2 values < or = 2.5 mm Hg and the survival status at 1 or 2 years after beginning of the therapy as endpoint. The highest positive and negative predictive values of all cut-off values were 0.50 and 0.41 at 1 year, and 0.81 and 0.26 at 2 years. CONCLUSIONS: Our data confirm the influence of the tumor oxygenation on the prognosis of patients with squamous cell carcinoma of the head and neck after radiotherapy. However, the calculated positive and negative predictive values suggest that the pO2 histography alone is not sufficient to be used as a predictive test to successfully select patients for treatment alternatives.

Virulence properties and serotypes of Shiga toxin-producing Escherichia coli from healthy Australian slaughter-age sheep.

A group of 1,623 ovine fecal samples recovered from 65 geographically distinct mutton sheep and prime lamb properties across New South Wales, Australia, were screened for the presence of Shiga toxin-producing Escherichia coli (STEC) virulence factors (stx(1), stx(2), eaeA, and ehxA). A subset was cultured for STEC isolates containing associated virulence factors (eaeA and/or ehxA), which were isolated from 17 of 20 (85%) and 19 of 20 (95%) tested prime lamb and mutton sheep properties, respectively. STEC isolates containing stx(1), stx(2), and ehxA were most commonly isolated (19 of 40 flocks; 47.5%), and this profile was observed for 10 different serotypes. Among 90 STEC isolates studied, the most common serotypes were O91:H(-) (22 isolates [24.4%]), O5:H(-) (16 isolates [17.8%]), O128:H2 (11 isolates [12.2%]), O123:H(-) (8 isolates [8.9%]), and O85:H49 (5 isolates [5.6%]). Two isolates (2.2%) were typed as O157:H(-). A total of 78 of 90 STEC isolates (86.7%) expressed Shiga toxin in Vero cell culture and 75 of 84 ehxA-positive isolates (89.3%) expressed enterohemolysin on washed sheep blood agar. eaeA was observed in 11 of 90 (12.2%) ovine STEC isolates, including serotypes O5:H(-), O84:H(-), O85:H49, O123:H(-) O136:H40, and O157:H(-). Although only 2 of 90 isolates were typed as O157:H(-), the predominant serotypes recovered during this study have been recovered from human patients with clinical disease, albeit rarely.

[Prognostic value of hemoglobin level for primary radiochemotherapy of head-neck carcinomas]. / Prognostischer Stellenwert des Hämoglobinwertes vor primärer Radiochemotherapie von Kopf-Hals-Karzinomen.

The pretherapeutic hemoglobin level (Hb) has been postulated to constitute a prognostic marker for outcome after primary chemoradiation of patients with advanced cancer of the head and neck. However, this hypothesis has not been tested systematically in large study samples. In the years 1992-1997, 125 patients with advanced head and neck cancer (stages III/IV UICC) were treated with primary chemoradiation in two different prospective multicentric trials, 62 patients in trial A (phase II, 1992-1995), and 63 in trial B (phase III, 1995-1997). Beside initial Hb, other pretherapeutic parameters with potential prognostic relevance were assessed and correlated with clinical outcome after 43-months follow-up: total tumor volume (TTV; calculated in initial CT scans), tumor oxygenation (polarographic measurements with Eppendorf histography), TNM, tumor localization, age, and performance status. The evaluation of the clinical end points (progression-free and overall survival and local tumor control) revealed that Hb and TTV were independent parameters with strong predictive character of outcome after primary chemoradiation in both trials (n = 125). Bivariate analysis showed < median (13.5 g/dl) a hazard ratio of 2.1 (P = 0.002) for Hb; and > median (98 ml) a Hazard ratio of 2.0 (P = 0.006) for TTV. Severe anemia (Hb < 10 g/dl) was an adverse factor in three patients. Hypoxia was associated with poorer initial therapeutical response but was not predictive of clinical outcome. Furthermore, tumor oxygenation showed no correlation with Hb. The other parameters examined failed to show prognostic significance. Our results indicate a high prognostic value of initial Hb for outcome after primary chemoradiation in advanced head and neck cancer and imply a therapeutic benefit of Hb substitution or erythropoietin administration. We propose to test this in randomized clinical trials.

Repeatability and prognostic impact of the pretreatment pO(2) histography in patients with advanced head and neck cancer.

PURPOSE: The purpose of this study was to evaluate the repeatability and the predictive relevance of the pretreatment pO(2) histography on the survival of patients with advanced head and neck cancer. PATIENTS AND METHODS: From July 1995 to August 1998, polarographic pO(2) measurements of lymph node metastases before therapy were performed in altogether 60 patients with histologically proven squamous cell carcinoma of the head and neck using the Eppendorf histograph. Forty-one of 60 patients were treated with an accelerated-hyperfractionated radiotherapy regimen with or without simultaneous chemotherapy as part of a multicenter phase III study. In 23 of 60 patients, two repeated independent measurements of the same tumor were performed with a time interval of approximately 24 h between the two measurements. RESULTS: The multivariate analysis revealed the fraction of pO(2) values

Oxygenation of advanced head and neck cancer: prognostic marker for the response to primary radiochemotherapy.

Recent clinical studies suggest that the degree of tumor oxygenation may be predictive of the response of radiation therapy for cancer. In an exploratory investigation of cervical lymph node metastases in 27 patients with advanced squamous cell carcinoma of the oropharynx and hypopharynx, this relationship was investigated by means of oxygen measurements with an Eppendorf PO (2) histograph. The measurements were made before the start of radiation therapy and after the first week of therapy. Clinical response was evaluated 6 weeks after the completion of therapy. Before therapy, marked hypoxia was observed in the lymph node metastases, with a mean PO (2) value of 16.1 +/- 8.2 mm Hg and a hypoxic fraction (PO (2) < 10 mm Hg) of 56.4% +/- 20.0%. After the first week of radiation (9 Gy) there was a general reoxygenation (DeltaPO (2) = 5.0 +/- 10.1 mm Hg, P < 0.05; Deltahypoxic fraction = -11.3% +/- 31.3%, P = 0.11). A relationship between the degree of reoxygenation and tumor response was not observed. Patients without at least partial lymph node response (n = 8) showed poorer pretherapeutic oxygenation (PO (2) mean = 11.1 +/- 2.9 mm Hg) than those who responded to the therapy (n = 19, PO (2) mean = 18.2 +/- 8.8 mm Hg). In this investigation of a defined set of patients with advanced carcinoma of the oropharynx and hypopharynx, we found that pretherapeutic oxygenation data are predictive for the therapeutic response to radiation therapy or radiochemotherapy.

Calcium dynamics and buffering in oculomotor neurones from mouse that are particularly resistant during amyotrophic lateral sclerosis (ALS)-related motoneurone disease.

Motoneurones are particularly vulnerable both in human forms of amyotrophic lateral sclerosis (ALS) and corresponding animal models of the disease. While most motoneurone populations are selectively impaired, oculomotor neurones are essentially resistant to ALS-related damage. Motoneurone vulnerability has been closely linked to disruptions of calcium signalling. To investigate underlying events, we performed a quantitative analysis of calcium homeostasis in oculomotor neurones from mice by simultaneous patch-clamp recordings in sliced tissue and microfluorometric-calcium measurements. Somatic calcium dynamics were investigated by using a computer-controlled microfluorometric system. In oculomotor neurones, basal calcium concentrations were around 80 nM and depolarisation-induced calcium responses were observed for membrane voltages positive to -40 u1u1u approximately mV1 approximately . Endogenous calcium homeostasis was quantified by using the 'added buffer' approach. The recovery phase of depolarisation-induced calcium transients was well approximated by a mono-exponential function with a decay time constant that showed a linear dependence on dye concentration. The extrapolated time constant in the absence of indicator dye was 1.7 +/- 0.2 s (n = 11 cells, 21C). Endogenous calcium binding ratios (kappa(s)) were found to be 264 +/- 25 (n = 11 cells), indicating that 99.6 % of cytosolic calcium ions were taken up by endogenous buffers. Recovery of calcium transients was characterised by an 'effective' extrusion rate gamma = 156 +/- 20 s-1 (n = 11 cells, 21 C). Endogenous calcium binding ratios in oculomotor neurones were 5- to 6-fold larger compared with those of more vulnerable motoneurones in the nucleus hypoglossus and spinal cord. In a first order approximation, they reduced the volume of local calcium elevations around open calcium channels, lowered peak amplitudes of global calcium transients for a given influx and prolonged calcium recovery times for a given set of uptake and extrusion mechanisms. With respect to motoneurone degeneration, our measurements suggest that the exceptional stability of oculomotor neurones partially results from a specialised calcium homeostasis based on high buffering capacities. Furthermore, they indicate that cellular adaptations that account for rapid calcium signalling in hypoglossal and spinal motoneurones enhance their vulnerability during ALS-related motoneurone disease.

Prognostic assessment of sonography and tumor volumetry in advanced cancer of the head and neck by use of doppler ultrasonography.

OBJECTIVE: The objective of this trial was to examine the degree of tumor vascularity in lymph node metastases as depicted by computer-assisted color Doppler sonography and the tumor volumes associated with prognosis in carcinomas of the oropharynx and hypopharynx after primary radiochemotherapy. PATIENTS AND METHODS: In a prospective trial, 25 patients with advanced squamous cell carcinomas of the oropharynx and hypopharynx (stage IV UICC 1997) were treated with radiochemotherapy. The color Doppler findings were quantified with a computer-assisted protocol that quantitatively describes color Doppler images by the relative color pixel density (CPD). As important prognostic cofactor, total tumor volume (TTV) was calculated from CT sections and related to the degree of vascularity. RESULTS: Low CPD in neck metastases showed a correlation with better overall survival. A high-CPD group and a high-TTV group (median survival 10.1 months) were determined and were compared with all other CPD/TTV combinations (median survival 28.4 months); the difference in survival was significant (P = 0.002). CONCLUSION: The results indicate that high tumor vascularity in combination with high TTV indicates a particularly bad prognosis in patients treated with primary radiochemotherapy for head and neck cancer.

Rise of oxygenation in cervical lymph node metastasis during the initial course of radiochemotherapy.

It has been hypothesized that during radiation treatment a reoxygenation of hypoxic tumor tissue takes place. To test this hypothesis, we have investigated whether reoxygenation in lymph node metastases could be determined by invasive PO (2) measurements. Through a hypodermic needle inserted transcutaneously into tumor-positive lymph nodes, polarographic oxygen determinations were made in 18 patients with advanced squamous cell carcinomas of the oropharynx and hypopharynx. These measurements were performed before therapy and a week after the onset of radiotherapy or radiochemotherapy, respectively. Low PO (2) values before treatment (mean value of the patient's median was 12.6 mm Hg PO (2)) and a mean hypoxic fraction (PO (2) < 5 mm Hg) of 39.6% indicated manifest tumor hypoxia. After 1 week of treatment, a significant increase in the median PO (2) (mean value of shift: 7.3 mm Hg) and a reduction in the hypoxic fraction (mean value of shift: 13.4% PO (2) < 5 mm Hg, P < 0.03) were observed after both radiotherapy and radiochemotherapy. Thus invasive PO (2) histography fulfills the requirements for a method to confirm tumor hypoxia in head and neck tumors. The results obtained indicate that reoxygenation occurs during the initial phases of radiotherapy and radiochemotherapy, and they will form the basis for future comparative investigations on the possible influence of hypoxic parameters on tumor responsiveness toward radiation and radiochemotherapy.

[Predictive value of serum levels of basic fibroblast growth factor, vascular endothelial growth factor and matrix metalloproteinase-2 in advanced carcinomas of the head and neck]. / Prognostische Bedeutung von bFGF, VEGF und MMP-2 im Serum von Patienten mit fortgeschrittenen Kopf-Hals-Tumoren.

The objective of this trial was to analyse the predictive character of the angiogenic factors vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and matrix metalloproteinase-2 (MMP-2) in the sera of patients with advanced carcinomas of the head and neck treated by primary radiochemotherapy. From 1992 to 1995, 25 patients with UICC stage cancers and one patient with stage III disease were treated in the departments of otolaryngology and radio-oncology of the University of Heidelberg according to a protocol of accelerated concomitant boost radiochemotherapy with carboplatin. The serum levels of VEGF, bFGF and MMP-2 were measured by enzyme-linked radiosorbent assay and data were correlated with followup findings (median time of follow up: 60 months). Patients with serum levels above normal were detected for VEGF in 4 patients, MMP-2 in 7 patients and for bFGF in 13 patients. An increase in bFGF serum levels above the upper limit of normal controls was significantly associated with a shorter time of locoregional control (P=0.036). In a covariate analysis bFGF proved to be independent of other prognostic factors, such as age, site, total tumor volume and response to therapy. No prognostic relevance of VEGF and MMP2 serum levels could be determined. The present suggest that bFGF is an independent prognostic factor for tumor control in advanced head and neck cancer after primary radiochemotherapy.