Cerebellar apparent diffusion coefficient changes in patients with supratentorial ischemic stroke.

OBJECTIVES: To evaluate apparent diffusion coefficient (ADC) in cerebellar subregions in patients with stroke. MATERIALS AND METHODS: The total counts and ADCs were bilaterally measured on cerebellar white matter, gray matters of medial (G1), intermediate (G2), and lateral zones (G3) on SPECT and ADC maps from 20 patients with supratentorial ischemic stroke within the first 48 h and on day 8 after onset. ADCs were also obtained from 15 age-matched controls. RESULTS: Within 48 h, the ADCs were significantly increased bilaterally in the G3, and tended to be increased bilaterally in the white matter and G1, and contralateral G2 compared with controls. On day 8, the ADCs were significantly increased in all contralateral cerebellar subregions and in ipsilateral G1 and G2, and tended to be increased in ipsilateral G3. The ADC value was significantly higher in contralateral than in ipsilateral white matter on day 8. The interhemispheric asymmetry indices (AIs) of ADC and SPECT were significantly associated with each other in G2 and G3 within 48 h, but not on day 8. The AIs of ADC and SPECT were significantly related to each other in the G3 within 48 h and on day 8. CONCLUSIONS: Supratentorial ischemic stroke may cause mild cerebellar vasogenic edema.

CD44 expression and its relationship with MMP-9, clinicopathological factors and survival in oral squamous cell carcinoma.

We investigated the expression of CD44 and MMP-9 in primary oral squamous cell carcinoma (OSCC) and evaluated their association with each other and clinicopathological factors as well as their prognostic value during long term follow up. Histological samples from 138 OSCC patients were immunohistochemically stained for the expression of CD44 and MMP-9. The staining results were compared with conventional prognostic factors and their impacts to patients' prognosis were also studied with survival analyses. Irregular staining of CD44 in tumour cells was associated with poor tumour differentiation (p=0.003), higher clinical stage (III-IV) (p=0.049), and the presence of T3-4 tumour stage (p=0.03). Strong stromal MMP-9 staining intensity was correlated with poor tumour differentiation (p=0.03). In univariate survival analysis irregular staining of CD44 in tumour cells was related to poor disease free and overall survival (p=0.001 and p<0.001, respectively). In multivariate analysis CD44 staining was a significant independent predictor for overall (p=0.03) and disease free survival (p=0.003). MMP-9 expression showed no statistical significance in survival analyses. Strong stromal staining intensity of MMP-9 correlated with irregular staining of CD44 in tumour cells, but had no prognostic significance in the present cohort. However, irregular staining of CD44 predicted more advanced disease and shortened survival of the patients.

Measurement of oedema in irritant-exposed skin by a dielectric technique.

BACKGROUND: Easily applicable water-specific instruments measuring local oedema in skin are not available. The aim of this study is to demonstrate quantitative assessment of skin oedema with the dielectric technique by measuring increase of skin water content related to sodium lauryl sulphate (SLS)-induced irritant contact dermatitis. METHODS: Irritant skin reaction and resulting oedema were induced by an irritant patch test on volar forearms in 12 healthy volunteers with the application of 1% SLS for 6 h. After occlusion the volunteers were divided into two groups: the patch test site of group I (six volunteers) received no treatment other than a base cream for the skin reaction, while for group II (six volunteers) a strong corticosteroid (clobetasol propionate) was applied on the irritant skin. During a follow-up of 72 h, erythema was scored visually, and irritant-induced oedema was measured with a novel water-specific instrument MoistureMeter-D. RESULTS: In the untreated irritant skin, a maximum increase of 45% in skin water content was found at 10 h postocclusion and water content was still elevated at 72 h. With these persons, the degree of oedema agreed well with the ultrasound-measured skin thickness (P=0.053). In the corticosteroid-treated skin, an increase of 8% in water content was measured during 72 h but there was no correlation between oedema and skin thickness. There was no correlation between erythema and oedema in untreated or corticosteroid-treated skin. CONCLUSION: The new instrument can easily be applied for noninvasive quantitative evaluation of local oedema and fluid retention in irritant-exposed skin.

ApoE polymorphism and acute stroke: a study with diffusion- and perfusion-weighted MRI and MR angiography.

OBJECTIVES: We examined whether the apolipoprotein E (ApoE) allele epsilon4 influences imaging findings in stroke as assessed by diffusion- (DWI) and perfusion-weighted (PWI) magnetic resonance imaging, and MR angiography (MRA). METHODS: Eight ApoE epsilon4 carriers and 15 non-carriers with acute ischemic stroke in the anterior circulation underwent DWI, PWI, and MRA within 24 h of stroke. DWI and PWI were repeated a week later. The apparent diffusion coefficient, relative cerebral volume (rCBV), relative cerebral blood flow (rCBF) and relative mean transit time were measured in three subregions on day one. RESULTS: In the ischemic core and the area of infarct growth, rCBV values were significantly higher in the epsilon4 carriers compared with the non-carriers. Based on the MRA findings, collateral blood flow was better in the epsilon4 carriers than in the non-carriers. Under the comparable severity of hypoperfusion, the hypoperfused area proceeded to infarction later or did not proceed to infarction at all in the non-carriers. CONCLUSION: These preliminary data suggest that in the ApoE allele epsilon4 carriers the threshold for the brain tissue to survive hypoperfusion versus to proceed to infarction seems to be different from that of the non-carriers.

Patient satisfaction with switching to Stalevo: an open-label evaluation in PD patients experiencing wearing-off (Simcom Study).

OBJECTIVES AND METHODS: This study investigated the ease with which 52 Parkinson's disease patients already receiving adjunct entacapone to traditional levodopa were switched to Stalevo (levodopa/carbidopa/entacapone). RESULTS: The switch to Stalevo was straightforward for most patients taking standard-release levodopa with 86% of these patients being able to replace their entire regimen without having to change the amount of levodopa taken. The majority of patients (54%, P = 0.162) preferred Stalevo; 31% preferred their prior treatment regimen; 15% had no preference. Patients found Stalevo more simple to dose (94%), more convenient to use (84%), easier to handle (84%), easier to remember (67%) and easier to swallow (59%), compared with their previous medication. CONCLUSIONS: Stalevo was well tolerated, with a low incidence of adverse events. The study shows that Stalevo is an effective, preferred and well-tolerated means of delivering levodopa/carbidopa/entacapone in one easy-to-use tablet.

Assessing the outcome of stroke: a comparison between MRI and clinical stroke scales.

OBJECTIVES: To assess the correlation of diffusion-weighted (DWI) and perfusion-weighted imaging (PWI) findings with the severity of acute neurologic deficit and their ability to predict short and long-term clinical outcomes of stroke. The ability of DWI and PWI to predict the outcome was compared with the ability of clinical stroke scales to predict the outcome. METHODS: Forty-eight patients with acute stroke underwent diffusion DWI and PWI on the first and eighth day after the ictus. Clinical and functional scales were carried out before each scan and 3 months after the stroke. RESULTS: The volumes of both the DWI and the PWI lesions correlated well with the acute neurologic deficit and the final outcome. The first day PWI (r = 0.64) and the National Institutes of Health Stroke Scale (NIHSS) scores (r = 0.70) correlated well with the final outcome. However, in logistic regression analysis, only the NIHSS score at the acute stage was the only independent predictor of the long-term clinical outcome. CONCLUSION: While the PWI and DWI lesion volumes correlated well with the outcome of the stroke, the imaging measurements did not improve the prognostic power over plain clinical stroke scale scores.

Versican expression in pharyngeal squamous cell carcinoma: an immunohistochemical study.

AIMS: To study the expression of versican, a large proteoglycan involved in repressing adhesion between cells and the extracellular matrix in pharyngeal squamous cell carcinoma (PSCC), and its relation to the expression of p53 and catenins, histological differentiation, clinical data, and prognosis. METHODS: For the retrospective survey, primary tumours for analyses were obtained from 118 patients diagnosed with PSCC of the oropharynx or hypopharynx. The immunohistochemical expression of versican was studied and was related to the expression pattern of p53 and catenins, in addition to clinical data and survival. RESULTS: In the primary tumours, strong stromal versican expression was graded as low in 59 (50%) and high in 59 (50%) cases. In addition, intracellular versican staining was seen in nine (8%) tumours. In local lymph node metastases, strong stromal versican staining was significantly more frequent compared with the primary tumours (p = 0.018). Strong stromal versican staining was more frequently seen in less advanced tumours (p = 0.015). There was no association between versican expression and the other investigated variables (p53, catenins, TNM status, and histological grade). Neither stromal nor intracellular versican expression predicted overall survival in these patients. CONCLUSIONS: Versican was more strongly expressed in the stroma of local metastases and in the earlier stages of disease in PSCC. However, versican expression was not an independent prognostic factor in this entity.

Validation of a new dielectric device to assess changes of tissue water in skin and subcutaneous fat.

Easily applicable and inexpensive water-specific techniques to evaluate local oedema, swollen tissue problems and fluid retention in humans are not available. In the present investigation a recently constructed non-invasive device for a local measurement of changes in tissue water in human skin and subcutaneous fat (SSF) was validated. The instrument transmits an ultra high-frequency electromagnetic (EM) wave of 300 MHz into a coaxial line and further into an open-ended coaxial probe which is in contact with the skin. Due to the dimensions of the applied probe the penetration of the EM field extends to subcutaneous fat. A major part of the EM energy is absorbed by tissue water while the rest is reflected back into a coaxial line. From the information of the reflected wave an electrical parameter, directly proportional to tissue water content, called a dielectric constant of SSF, was calculated. For system validation, the decrease of water content in SSF measured with the dielectric technique in the volar forearm of seven patients during haemodialysis treatment was compared with the decrease of the circumference of the forearm and the amount of fluid removed. Statistically highly significant correlations were obtained between the decreasing dielectric constant (i.e. water content) of the SSF and the fluid removed during haemodialysis treatment (r = -0.99, p < 0.01) and between the decreasing dielectric constant and the circumference of the arm (r = 0.97, p < 0.05). The sensitivity of the dielectric method was four-fold compared with the circumferential measurement. The repeatability 3.0% was not dependent on the phase of haemodialysis. The new device allows an easy and non-invasive measurement technique to assess changes of tissue water in SSF.

Reduced expression of hyaluronan is a strong indicator of poor survival in oral squamous cell carcinoma.

Several malignant tumours accumulate hyaluronan (HA), a matrix component suggested to promote cancer cell growth and migration. The expression and prognostic value of HA was analysed in a cohort of 151 oral squamous cell carcinoma (SCC) patients with adequate archival tumour material and follow-up data. The tumour samples were stained using a biotinylated HA-specific probe. Normal squamous epithelium showed a strong and homogeneously distributed staining for HA. The most superficial layers were HA-negative. In moderate (n=11) and high grade (n=16) dysplasias an irregular HA staining was observed around invasive cancer. Malignant transformation in oral squamous cell epithelium changed the staining toward irregular with focal reduction of HA. The well (n=92) or moderately differentiated (n=47) carcinomas had a strong HA staining intensity. In poorly differentiated tumours (n=12) the HA staining was weaker and mainly intracellular. The stromal tissue showed usually moderate (n=69) or strong (n=67) HA staining intensity with no statistically significant correlation with the degree of tumour differentiation. At the end of the follow-up (median 52 months) 66 (43%) patients had died because of an oral SCC. A significant difference in overall survival (OS) and disease free survival (DFS) (P=0.0002 and 0.0020, respectively) was noticed between the patients with the different epithelial staining patterns for HA. The reduction of HA staining was associated with poor survival. In Cox's multivariate analysis HA staining was a significant independent predictor of OS (P=0.011) and DFS (P=0.013). These results suggest that HA is a prognostic marker in oral squamous cell carcinoma.

Changes in abdominal subcutaneous fat water content with rapid weight loss and long-term weight maintenance in abdominally obese men and women.

OBJECTIVE: Insulin resistance decreases blood flow and volume in fat tissue. We hypothesised that fat tissue nutritive blood flow and volume, and thereby water content, would increase during weight loss and weight maintenance in obese persons. DESIGN: Longitudinal clinical intervention with a 9-week very-low-calorie diet (VLCD) followed by one year of weight maintenance. SUBJECTS: Obese men (n=13) and women (n=14) with the metabolic syndrome. MEASUREMENTS: Water content of abdominal subcutaneous fat tissue as estimated by a sensor on the skin surface measuring the dielectric constant at 300 MHz. Anthropometric measures of fatness and fat distribution. Biochemical measures related to insulin resistance. RESULTS: Subjects lost 14.5+/-3.4% of body weight during the VLCD, and generally sustained this weight loss during weight maintenance. Insulin sensitivity as estimated by an index (qualitative insulin sensitivity check index) increased during the VLCD, and remained increased throughout weight maintenance. The dielectric constant increased from 23.3+/-2.3 to 25.0+/-2.1 (P<0.001) during the VLCD, and further to 27.8+/-1.9 (P<0.001) during weight maintenance, indicating an increase in the water content of subcutaneous fat. The increase in subcutaneous fat water content did not correlate with weight loss and other measures of adiposity during the VLCD, but there was an inverse correlation that strengthened in significance from baseline to 6, 9 and 12 mo (r=-0.32 to -0.64, P=0.079-0.002). Increases in subcutaneous fat water content also correlated with improvements in insulin sensitivity at 6, 9 and 12 months of weight maintenance (r=0.34-0.54, P=0.094-0.006). CONCLUSIONS: Water content of abdominal subcutaneous adipose tissue increases with weight loss in obese persons with the metabolic syndrome, and may reflect increased subcutaneous fat tissue nutritive blood flow. The increase in water content correlates with the increase in insulin sensitivity, suggesting that weight loss and consequent improved insulin sensitivity could mediate the increase in abdominal subcutaneous fat hydration.

Detecting the subregion proceeding to infarction in hypoperfused cerebral tissue: a study with diffusion and perfusion weighted MRI.

Diffusion and perfusion weighted MRI have been widely used in ischaemic stroke. We studied 17 patients in whom ischaemic areas showed an ischaemic core, an area of infarct growth and hypoperfused but ultimately surviving tissue. Apparent diffusion coefficients (ADC) were measured on days 1, 2, and 8 in the three subregions and in contralateral control areas. Cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) were measured in these regions on day 1 perfusion maps. On day 1, the ischaemic core had very low ADC and CBF and increased MTT. The ADC in the ischaemic core gradually increased during the week. The area of infarct growth on day 1 had slightly but significantly decreased ADC (96% of control, P=0.028), moderately decreased CBF and increased MTT. On day 1 the hypoperfused but surviving tissue had slightly but significantly increased ADC (103% of control, P=0.001), mildly decreased CBF and increased CBV and MTT. The ADC of the area of infarct growth decreased to the same level as in the ischaemic core on days 2 and 8. That of surviving tissue was still above normal on day 2 (103% of control), but had returned to the normal level by day 8. Measurement of ADC combined with perfusion MRI may help distinguish different subregions in acutely hypoperfused brain.

Colored moderately thermophilic bacteria in paper-machine biofilms.

Biofilms cause several problems in papermaking. This report describes a microbiological survey of colored biofilms in six paper and board machines, including two case studies of outbreaks of colored slimes in which the causative bacteria were found. A total of 95 pink-, red-, orange- or yellow-pigmented strains were isolated. Nearly all (99%) of the strains grew at 52 degrees C, 72% grew at 56 degrees C, but only 30% grew at 28 degrees C, indicating that most of the strains were moderately thermophilic. Biofilm formation potential and biocide susceptibility of the strains were analyzed with a microtiter plate assay. In the presence of 5 ppm of methylene bisthiocyanate or 2,2-dibromo-3-nitrilopropionamide in paper-machine water, 55 strains formed biofims. Moreover, 39 strains increased biofilm production by 5-753% in the presence of biocide, suggesting that biocide concentrations inhibitory to planktonic but not to surface-attached cells may actually promote biofouling. The cells may have inactivated a portion of the biocides, as the cell density in this assay was high, corresponding to the highest cell densities occurring in the circulating waters. Four groups of colored bacteria that were isolated from several mills were identified. Pink-pigmented Deinococcus geothermalis and red-pigmented Meiothermus silvanus occurred as common primary biofilm-formers in paper machines. This report is the first description of the involvement of Meiothermus species in red-slime formation in the paper industry. The third group of bacteria (putative new species related to Roseomonas) contained strains that were not biofilm formers, but which were commonly found in slimes of neutral or alkaline machines. The fourth group contained red-pigmented biofilm-forming strains representing a novel genus of alpha- Proteobacteria related to Rhodobacter. Many colored paper-machine bacteria are species previously known from microbial mats of hot springs. Some characteristics of the bacterial groups are described here in order to facilitate their recognition in future cases of colored-slime outbreaks in the paper industry.

Thermomonas haemolytica gen. nov., sp. nov., a gamma-proteobacterium from kaolin slurry.

Four aerobic, gram-negative bacterial strains isolated from kaolin slurry used in the production of paper were subjected to a polyphasic analysis and characterization to determine their taxonomic position. Analysis of the 16S rDNA sequences of the four strains revealed that they represent a new lineage within the gamma-Proteobacteria, related to the genera Xanthomonas, Pseudoxanthomonas, Stenotrophomonas, Luteimonas, Xylella and Rhodanobacter. Analysis of the quinone system, the polyamines, the fatty acids and the polar lipids revealed a combination of characteristics that is unique and not described for the phylogenetic relatives. The four strains contain a ubiquinone Q-8, spermidine as the major polyamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as the predominant polar lipids, and a fatty acid profile with predominantly iso-branched fatty acids. The G+C content of the genomic DNA was determined to be within the narrow range 67.1-68.7 mol%. Determination of DNA relatedness, as well as riboprint band patterns and amplified fragment length polymorphism profiles, clearly demonstrated that the four strains are members of a single species. Antibiotic-susceptibility patterns were identical for the four strains. Although showing a high degree of similarites in physiological and biochemical patterns, each of the four strains could be distinguished from the others on the basis of a few biochemical characteristics. On the basis of the estimates of phylogenetic relationships derived from the 16S rDNA sequence analyses, the observed chemotaxonomic characteristics and other phenotypic traits, a new genus, Thermomonas gen. nov., and species, Thermomonas haemolytica sp. nov., are proposed for the strains A50-7-3T (= DSM 13605T = LMG 19653T), B 50-7-1 (= DSM 13598 = LMG 19655), D50-7-1 (= DSM 13610 = LMG 19656) and B50-8-1 (= DSM 13599 = LMG 19654), with strain A50-7-3T as the type strain.

Mass spectrometric studies on small open-chain piperazine-containing ligands and their transition metal complexes.

Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry was used to characterize the complexes formed between open-chain piperazine-containing ligands and transition metal salts (Cobalt, Copper, Zinc, and Cadmium as chlorides, nitrates, and acetates). Only single-charged complexes were observed, formed of one ligand (L) and mainly one metal ion (M). Since the net charge of the complexes was one, a counterion (X) was attached to some of the complexes, with formation of [L + M + X]+ complexes, and a proton was lost from others, as in [L - H + M]+ complexes. In most cases the composition of the complexes was more dependent on the ligand than the metal salt. Collision-induced dissociation measurements showed that complexes with related composition often differed in structure, or that interactions between the ligand and the metal ion were not alike. The metal ion influenced considerably the fragmentation pathways of the ligands, so that the fragmentation products could be used to deduce the binding sites of the metal. The variations observed in fragmentation behavior of complexes possessing the same ligand but different metal ions can mostly be explained by the ionic radius and electronic configuration of the metal ion. The results indicated a preference of the piperazine ring of the coordinated ligand for the boat conformation.

Mass spectrometric studies on pyridine-piperazine-containing ligands and their complexes with transition metals formed in solution.

Electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) methods were used to study open-chain piperazine-containing ligands (L) and their complexes formed with transition-metal salts. ESI and MALDI measurements were performed with a Fourier transform ion cyclotron resonance (FT-ICR) and a time-of-flight (TOF) mass spectrometer, respectively. Only singly charged complexes, between one ligand and one or several metal ions, were formed in the ESI measurements. Because the net charge was always one, one or several counterions were attached to the complex. Under ESI conditions, the complexes formed between the ligands and metal (Co, Ni, Cu, and Cd) salts were [L + M + X](+), [L + H + M + X(2)](+) and [L + M(2) + X(3)](+) (M = metal ion, X = counterion). In collision induced dissociation reactions the [L + H + M + X(2)](+) complexes easily eliminated one proton and one counterion. Fragmentation pathways were more dependent on the metal ion than the ligand, and elimination of the second counterion occurred with one proton from copper and nickel complexes and with one proton and one hydrogen from cobalt complexes. Differences in the fragmentation of the complexes could be due to electronic configuration of the metal ion. In the MALDI measurements the ratio between the [L + H](+) and [L - H](+) ions varied with the matrix. Fragmentation of the ligands through elimination of 2-methylpyridine end groups occurred with the aromatic matrices containing carboxylic acid and hydroxyl substituents. Ionization of the complexes was not successful with MALDI as the matrix molecules were also attached to the complexes.

In vitro and in vivo responses to the recombinant bovine dander allergen Bos d 2 and its fragments.

BACKGROUND: About one in every four cases of occupational rhinitis recorded in Finland is animal-induced. Bovine allergens are the most important in this respect and the largest patient group consists of dairy farmers. Allergen immunotherapy, if proven effective, safe and feasible, would be ideal for their treatment. The development of recombinant allergens has offered new potential therapeutic prospects. Fragments of recombinant Bos domesticus (Bos d 2) allergen could be suitable for this purpose because they are recognized by T cells but their IgE-binding capacity is attenuated. OBJECTIVE: The aim of this study was to verify whether the potential of the two fragments of recombinant Bos d 2 (corresponding to amino acids 1-131 and 81-172) to induce immediate allergic reaction in a shock organ (nose) was decreased compared to the complete recombinant allergen, which would be an advantageous property for a preparation intended for allergen immunotherapy. METHODS: The study group consisted of 10 dairy farmers with cow-induced allergic rhinitis. We used the IgE titres against native Bos d 2 measured by indirect IgE ELISA to characterize the level of sensitization and compared the IgE titres in the rhinitis patients with 12 cow-sensitized asthmatic farmers and 12 healthy students. In vitro reactivity against recombinant Bos d 2 and its two fragments was studied by indirect IgE ELISA and in vivo reactivity by nasal provocation tests. RESULTS: The IgE titres against native Bos d 2 of patients with rhinitis tended to be lower than the titres of asthmatics. The healthy students did not exhibit any detectable IgE reactivity to native Bos d 2. In the patients with rhinitis, there was no statistically significant difference between IgE responses against native and recombinant Bos d 2, whereas with both in vitro and in vivo, the reactivity to both fragments of recombinant Bos d 2 was lower than the reactivity to the complete recombinant allergen. CONCLUSIONS: Due to the decreased in vivo capacity to induce immediate allergic reactions, the fragments may be better tolerated in allergen immunotherapy than the complete allergen.

Diffusion and perfusion MR imaging in acute ischemic stroke: a comparison to SPECT.

Diffusion (DWI) and perfusion (PWI) magnetic resonance imaging are relatively new methods of clinical imaging that probably can detect infarcted (DWI) and hypoperfused but still salvageable tissue (PWI) in acute human stroke. Forty-six acute stroke patients were imaged within 24 h of ictus, on the second day and after a week. SPECT was also performed on 23 patients in the acute phase (first or second day). On the first day, mean volume of hypoperfused tissue was significantly greater (P<0.001) than the infarcted tissue. The initial hypoperfusion volume correlated significantly with the final infarct size (P<0.001). The initial perfusion-diffusion mismatch correlated significantly with the infarct growth (P< or =0.001). The hypoperfusion volumes measured from PWI and SPECT correlated significantly (P<0.001). In conclusion, combined DWI and PWI is a powerful tool in evaluating the hemodynamics of acute ischemic stroke and can predict the infarct growth during 1 week.

Paranasal sinus mucormycosis: a report of two cases.

Mucormycosis of the nose and paranasal sinuses is a rare invasive fungal infection, which often has a very fulminant course and characteristic clinical findings. The patients are usually immunocompromised, with diabetic ketoacidosis being the commonest underlying disorder. In some immunocompetent patients, the disease is associated with local predisposing factors, such as chronic sinusitis. Although the prognosis has improved in recent decades, the disease can still be fatal. The underlying disease is an important determinant of prognosis and correction of the metabolic disorder, if present, is essential. Herein we report two cases: one of our patients was immunocompetent but had earlier suffered from polypous rhinosinusitis whereas the other had mild adult-type diabetes. Both patients were successfully treated with surgical debridement and amphotericin B.

Mechanisms of biofilm formation in paper machine by Bacillus species: the role of Deinococcus geothermalis.

Mechanisms for the undesired persistence of Bacillus species in paper machine slimes were investigated. Biofilm formation was measured for industrial Bacillus isolates under paper machine wet-end-simulating conditions (white water, pH 7, agitated at 45 degrees C for 1-2 days). None of the 40 tested strains of seven Bacillus species formed biofilm on polished stainless steel or on polystyrene surfaces as a monoculture. Under the same conditions, Deinococcus geothermalis E50051 covered all test surfaces as a patchy thick biofilm. The paper machine bacilli, however, formed mixed biofilms with D. geothermalis E50051 as revealed by confocal microscopy. Biofilm interactions between the bacilli and the deinococci varied from synergism to antagonism. Synergism in biofilm formation of D. geothermalis E50051 was strongest with Bacillus coagulans D50192, and with the type strains of B. coagulans, B. amyloliquefaciens or B. pumilus. Two B. licheniformis, one B. amyloliquefaciens, one B. pumilus and four B. cereus strains antagonized biofilm production by D. geothermalis. B. licheniformis D50141 and the type strain of B. licheniformis were the strongest antagonists. These bacteria inhibited deinococcal growth by emitting heat-stable, methanol-soluble metabolite(s). We conclude that the persistence of Bacillus species in paper machine slimes relates to their ability to conquer biofilms formed by primary colonizers, such as D. geothermalis.