Butyricimonas virosa bacteraemia and bowel disease: case report and review.

Only two cases of human infection with the anaerobic Gram-negative bacillus Butyricimonas virosa have been previously reported. We describe the case of a 69-year-old man with B. virosa and diverticulitis, further supporting an association of bacteraemia with this pathogen to bowel disease. We also summarize the characteristics of the previously described cases.

Effects of diet-matrix on volatile organic compounds in breath in diet-induced obese mice.

Breath gas analysis in humans proved successful in identifying disease states and assessing metabolic functions in a non-invasive way. While many studies report diagnostic capability using volatile organic compounds (VOC) in breath, the inter-individual variability even in healthy human cohorts is rather large and not completely understood in its biochemical origin. Laboratory mice are the predominant animal model system for human disorders and are analysed under highly standardized and controlled conditions. We established a novel setup to monitor VOCs as biomarkers for disease in the breath gas of non-anesthetized, non-restrained mice using a proton transfer reaction mass spectrometer with time of flight detection. In this study, we implemented breath gas analysis in a dietary intervention study in C57BL/6J mice with the aim to assess the variability in VOC signatures due to a change in the diet matrix. Mice were fed a standard laboratory chow and then exposed to four semi-purified low- or high-fat diets for four weeks. Random forest (RF++) was used to identify VOCs that specifically respond to the diet matrix change. Interestingly, we found that the change from a chow diet to semi-purified diets resulted in a considerable drop of several VOC levels. Our results suggest that the diet matrix impacts VOC signatures and the underlying metabolic functions and may be one source of variability in exhaled volatiles.

Influences of mixed expiratory sampling parameters on exhaled volatile organic compound concentrations.

Breath gas analysis is a promising technology for medical applications. By identifying disease-specific biomarkers in the breath of patients, a non-invasive and easy method for early diagnosis or therapy monitoring can be developed. In order to achieve this goal, one essential prerequisite is the reproducibility of the method applied, i.e. the quantification of exhaled volatile organic compounds (VOCs). The variability of breath gas VOC measurements can be affected by many factors. In this respect, sampling-specific parameters like flow rate and volume of exhalation, exhalation with or without breath holding, exhalation in single or multiple breathing and volume of air inhaled before breath gas exhalation can play a vital role. These factors affecting the measurements must be controlled by optimizing the sampling procedure. For such an optimization, it is important to know how exactly the different parameters affect the exhaled VOC concentrations. Therefore, a study has been undertaken in order to identify some effects of different breath sampling-specific parameters on the exhaled VOC profile using the mixed expired breath sampling technique. It was found that parameters such as filling the sampling bag with high or low flow rate of exhalation, with multiple or single exhalations, in different volumes of exhalation, with breath holding and under different surrounding air conditions significantly affect the concentrations of the exhaled VOCs. Therefore, the specific results of this work should be taken into account before planning new breath gas studies or developing new breath gas collection systems in order to minimize the number of artefacts affecting the concentration of exhaled VOCs.

Headspace measurements of irradiated in vitro cultured cells using PTR-MS.

A pilot study was performed to evaluate a new concept for a radiation biodosimetry method. Proton transfer reaction-mass spectrometry (PTR-MS) was used to find out whether radiation induces changes in the composition of volatile organic compounds (VOCs) in the headspace of in vitro cultured cells. Two different cell lines, retinal pigment epithelium cells hTERT-RPE1 and lung epithelium cells A-549, were irradiated with gamma radiation at doses of 4 Gy and 8 Gy. For measuring the cell-specific effects, the VOC concentrations in the headspace of flasks containing cells plus medium, as well as of flasks containing pure medium were analyzed for changes before and after irradiation. No significant radiation-induced alterations in VOC concentrations in the headspace could be observed after irradiation.

Discrimination of cancerous and non-cancerous cell lines by headspace-analysis with PTR-MS.

Proton transfer reaction mass spectrometry (PTR-MS) has been used to analyze the volatile organic compounds (VOCs) emitted by in-vitro cultured human cells. For this purpose, two pairs of cancerous and non-cancerous human cell lines were selected:1. lung epithelium cells A-549 and retinal pigment epithelium cells hTERT-RPE1, cultured in different growth media; and 2. squamous lung carcinoma cells EPLC and immortalized human bronchial epithelial cells BEAS2B, cultured in identical growth medium. The VOCs in the headspace of the cell cultures were sampled: 1. online by drawing off the gas directly from the culture flask; and 2. by accumulation of the VOCs in PTFE bags connected to the flask for at least 12 h. The pure media were analyzed in the same way as the corresponding cells in order to provide a reference. Direct comparison of headspace VOCs from flasks with cells plus medium and from flasks with pure medium enabled the characterization of cell-line-specific production or consumption of VOCs. Among all identified VOCs in this respect, the most outstanding compound was m/z = 45 (acetaldehyde) revealing significant consumption by the cancerous cell lines but not by the non-cancerous cells. By applying multivariate statistical analysis using 42 selected marker VOCs, it was possible to clearly separate the cancerous and non-cancerous cell lines from each other.

The effects of low environmental cadmium exposure on bone density.

Recent epidemiological data indicate that low environmental exposure to cadmium, as shown by cadmium body burden (Cd-U), is associated with renal dysfunction as well as an increased risk of cadmium-induced bone disorders. The present study was designed to assess the effects of low environmental cadmium exposure, at the level sufficient to induce kidney damage, on bone metabolism and mineral density (BMD). The project was conducted in the area contaminated with cadmium, nearby a zinc smelter located in the region of Poland where heavy industry prevails. The study population comprised 170 women (mean age=39.7; 18-70 years) and 100 men (mean age=31.9; 18-76 years). Urinary and blood cadmium and the markers of renal tubular dysfunction (beta(2)M-U RBP, NAG), glomerular dysfunction (Alb-U and beta(2)M-S) and bone metabolism markers (BAP-S, CTX-S) as well as forearm BMD, were measured. The results of this study based on simple dose-effect analysis showed the relationship between increasing cadmium concentrations and an increased excretion of renal dysfunction markers and decreasing bone density. However, the results of the multivariate analysis did not indicate the association between exposure to cadmium and decrease in bone density. They showed that the most important factors that have impact on bone density are body weight and age in the female subjects and body weight and calcium excretion in males. Our investigation revealed that the excretion of low molecular weight proteins occurred at a lower level of cadmium exposure than the possible loss of bone mass. It seems that renal tubular markers are the most sensitive and significant indicators of early health effects of cadmium intoxication in the general population. The correlation of urinary cadmium concentration with markers of kidney dysfunction was observed in the absence of significant correlations with bone effects. Our findings did not indicate any effects of environmental cadmium exposure on bone density.

Hydrophilic olive cake extracts: characterization by physicochemical properties and Cu(II) complexation.

Disposed olive cake generates hydrophilic components that can be mobilized in the aquatic environment. This paper deals with the characterization of such components, isolated by alkaline extraction. It is shown that these substances possess properties very much resembling humic acid, including a substantial inventory of proton exchanging groups. Extraction and purification of the hydrophilic components from the disposed olive cake was performed by the standard approach for isolation of humic acids from solid sources, i.e. alternating alkaline dissolution and acid flocculation, leaving the purified extract in the protonated form. The purified sample was characterized by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), Fourier Transform Infra Red Spectroscopy (FTIR), UV-vis, Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) and Asymmetrical Flow Field-Flow Fractionation (AFFFF). The complex formation properties were investigated by potentiometry using Cu(II) ion selective electrode under atmospheric conditions at I=0.1M NaClO(4) (aqueous solution) and pH 6. The formation constant for the CuHA complex is found to be logbeta=5.3+/-0.4 which is close to the corresponding value (logbeta=5.2+/-0.4) obtained from similar investigations with the commercially available Aldrich humic acid (this study) and corresponding published values for various humic acids. Both, structural properties and complex formation data show that the olive cake extract has considerable similarities with humic acids from different sources, pointing towards potential similarities in environmental behavior and impact.

Investigations on the variability of breath gas sampling using PTR-MS.

Breath gas analysis is a promising technology in the frame of medical diagnostics. By identifying disease-specific biomarkers in the breath of patients, a non-invasive and easy method for early diagnosis or therapy monitoring might be developed. However, to verify this potential and develop diagnostic tools based on breath gas analysis one essential prerequisite is a low variability in measurement of exhaled volatile organic compounds. Therefore, a study has been undertaken in order to identify possible artefacts within the application of a breath gas test in practice, for which the breath gas is analysed by proton transfer reaction-mass spectrometry (PTR-MS). After validating the low instrumental variability by repeatedly measuring standard gas, the variability of breath gas sampling has been evaluated. The latter has been carried out by measuring single breath gas samples (mixed expiratory breath) collected over different periods of time such as 1 min (10 volunteers, 4 breath gas samples each), 1 h (10 volunteers, 11 breath gas samples each) and several days (11 volunteers, 10 breath gas samples each). The breath gas samples were collected in Teflon bags and consecutively measured with PTR-MS. It was found that those samples collected within 1 min and 1 h show a low variability. This was, however, not the case for samples being collected over longer periods of time (15-70 days). Under these circumstances, many volatile organic compounds (VOCs) showed significant day-to-day variation in concentration, although the breath collection had been performed under the same conditions (similar sampling time, sampling technique, sample storage time, measurement conditions, etc). This large variation might be assigned to the influence of room air VOCs, which have been investigated in this work, or with other parameters which will be discussed. It was also found that the variability in the measurement of exhaled concentrations of methanol, acetone and isoprene within different individuals (inter individual variability) is much higher than differences in the same volunteer (intra individual variability) measured over a longer time interval.

Does fecal diversion offer any chance for spontaneous closure of the radiation-induced rectovaginal fistula?

Analysis of the clinical course of patients with postirradiation rectovaginal fistula after fecal diversion. The studied group included 17 women with postirradiation rectovaginal fistula who underwent fecal diversion as a sole mode of treatment, between January 1987 and December 2002, in our department. All patients were subjected to radiotherapy due to cancer of the uterine cervix, administered 5-107 months before the fistula appearance (mean, 22.9 months). In 3 of 17 patients (18%), spontaneous closure of fistula was observed after 5, 6, and 9 months, respectively, from fecal diversion. Closure was confirmed by endoscopy. Length of follow-up after fecal diversion ranged from 0.5 to 122 months. The actuarial probability of spontaneous closure of postradiotherapy rectovaginal fistula was 0.24 at 9 months of follow-up and then remained stable thereafter. In conclusion, colostomy alone gives hardly a chance for closure of the postradiotherapy rectovaginal fistula. Additional surgical measures are necessary.

Expression of tumour necrosis factor-related apoptosis-inducing ligand and caspase-3 in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

AIM: To assess whether the distribution of the recently described proapoptotic ligand, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), and the apoptosis effector, caspase-3 alters with the degree of inflammation and fibrosis present in liver biopsy specimens from patients with chronic hepatitis C virus infection. METHODS AND RESULTS: Expression of TRAIL and caspase-3 was assessed immunohistochemically in liver biopsy specimens obtained from 89 adults with chronic hepatitis C. Expression of TRAIL in hepatocytes correlated inversely with stage of fibrosis (P = 0.001), classified according to the Scheuer score; expression of caspase-3 in hepatocytes correlated with grade of inflammation (P = 0.012). Expression of TRAIL in hepatocytes was not correlated with grade of inflammation (P > 0.05); expression of caspase-3 was not correlated with stage of fibrosis (P > 0.05). Maximum expression of proapoptotic TRAIL protein was observed in cases with low grade inflammation (G0) and low stage fibrosis (S1). Maximum expression of caspase-3 in hepatocytes was observed in cases with high grade inflammation (G3-4) and high stage fibrosis (S3), but not with liver cirrhosis (S4). CONCLUSIONS: There is a significant decrease in TRAIL expression with increasing grade of inflammation, whereas caspase-3 expression is significantly increased with advanced fibrosis, short of cirrhosis.

Expression of Bax, Bcl-xL and Bcl-2 proteins in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

AIMS: To determine the expression of regulators of apoptosis in chronic hepatitis C. METHODS AND RESULTS: Expression of Bax, Bcl-xL and Bcl-2 proteins was assessed immunohistochemically in liver biopsy specimens obtained from 89 adults with chronic hepatitis C. Expression of Bax in hepatocytes correlated inversely with grade of inflammation (P < 0.001) and stage of fibrosis (P = 0.011), classified according to the Scheuer score; expression of Bcl-xL in hepatocytes did not correlate with grade of inflammation (P = 0.106) or stage of fibrosis (P = 0.078); maximum Bcl-xL expression was observed in grade 3 inflammation and stage 4 fibrosis. Expression of Bcl-2 protein in hepatocytes was present in only two cases (both with advanced disease); the expression of Bcl-2 protein in interlobular bile duct epithelial cells correlated with the grade of inflammation (P = 0.018), but not with stage of fibrosis (P = 0.154). The expression of Bcl-2 protein in lymphoid cells infiltrating portal zones and lobules did not correlate with grade of inflammation (P = 0.113) or stage of fibrosis (P = 0.815). CONCLUSION: Major differences in expression of studied proteins were observed in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

Patients with Paget disease of nipple and with palpable mass in breast have unfavorable prognosis.

Recent data indicate that in breast cancer patients the presence of Paget disease of the nipple may be related to poor prognosis. Therefore, we decided to assess long-term results of the treatment of such patients, and to assess the relationship between the physical and pathological findings and prognosis. The files of 60 patients with Paget disease of the nipple who were treated between 1977 and 2000 were analyzed retrospectively with respect to the results of physical and pathologic examinations, disease recurrence and survival. In 38/60 patients, the cancer was invasive. In 26/60 patients, palpable masses in the breast were diagnosed. The 5-year overall survival probability was 0.68; the probability was 0.82 for patients without palpable masses, 0.51 for those with palpable masses, 0.91 for patients without invasive cancer and 0.58 for patients with associated invasive cancer. In conclusions, patients with Paget disease of the nipple and with palpable mass in the breast had unfavorable diagnosis. The nature of all nipple changes should be explained as early as possible in order to diagnose the disease when no mass is palpable in the breast.

Glutathione S-transferase M1 and P1 metabolic polymorphism and lung cancer predisposition.

Individual susceptibility to different environmental agents is expected to be associated with alterations in metabolism of xenobiotics. Thus, genetic polymorphism of glutathione S-transferase (GST) can be recognized as a potential risk modifier in lung cancer development. The distribution of GSTM1 and GSTP1 genotypes was studied in a group of 138 diagnosed lung cancer patients and in 165 controls living in central Poland and RFLP-PCR technique was applied. The frequency of GSTM1 null genotype and GSTP1 Val single and duplicated alleles was similar among patients and controls. GSTM1 homozygous deletion was most prevalent in small-cell carcinoma groups (adjusted odds ratio (OR): 2.32, 95% confidence interval (CI): 0.98-5.52). In patients and controls, GSTM1A genotype was most frequent (34.1% vs. 37.0%). The estimated lung cancer risk for GSTM1 null, GSTP1 Ile/Val and GSTP1 Val/Val combined genotype was 1.44 (95% CI: 0.73-2.83), suggesting the absence of modifying effect of defective GSTM1 and GSTP1 alleles on lung cancer predisposition.

Immunohistochemical evaluation of P21ras and P53 proteins expression in human non-small-cell lung cancers.

Human non-small-cell lung cancers (NSCLCs) of 48 patients were analyzed immunohistochemically to detect P21 ras and P53 proteins expression. The relationship between P21 ras and P53 proteins expression and clinicopathologic findings was also assessed. DAKO EnVision TM detection system was employed in the study. The P21 ras and P53 proteins expression was shown in 75% (36/48) and 33.3% (16/48) studied NSCLCs, respectively. In both cases the difference was significant when compared with adequate negative control. Simultaneous expression of both studied proteins was observed in all cases in which P53 expression was noticed. No significant association of P21 ras and P53 expression was found with age, histologic type, histologic grade, tumor size or lymph node metastasis of the studied NSCLCs. Therefore, our study suggests that P21 ras and P53 protein play a role in the pathogenesis of NSCLCs but they have no value as a prognostic markers in the case of lung cancers.

Hearing loss among workers exposed to moderate concentrations of solvents.

OBJECTIVES: It is known that some industrial organic solvents are ototoxic. This study was aimed at evaluating the hearing effects of a mixture of organic solvents alone or in combination with noise on employees in paint and lacquer enterprises. The concentration of solvents was below the occupational exposure limits (OEL) for most of the subjects. METHODS: Altogether 517 subjects were divided into the following three groups: persons with no risk due to noise or organic solvent exposure at the workplace, workers exposed to organic solvents only, and workers exposed to both organic solvents and noise. RESULTS: The relative risk (RR) of hearing loss in the solvent-only exposure group was significantly increased (RR 4.4 and RR 2.8 for noise exposure of < 80 dB-A and < 85 dB-A, respectively) in a wide range of frequencies (2-8 kHz). No additional risk in the solvent + noise exposure group was found (RR 2.8). Hearing thresholds were significantly poorer in a wide range of frequencies (1-8 kHz) for both groups exposed to solvents, when compared with the reference group. The mean hearing thresholds at frequencies of 2-4 kHz were poorer for workers exposed to solvents + noise than for the solvent-only group; this finding suggests an additional effect for noise. However, there was no correlation between hearing loss and the extent of solvent exposure. CONCLUSIONS: The results indicate that occupational organic solvent exposure at moderate concentrations increases the risk of hearing loss, and the ototoxic effects should be considered when the health effects of exposed workers are monitored.

Nested case-control study of lung cancer among pulp and paper workers in relation to exposure to dusts.

BACKGROUND: Numerous studies have indicated an increased risk of lung cancer in pulp and paper industry workers. In a 1990 survey, standardized mortality ratio (SMR) was found to be 122 (95% CI:96-153) for lung cancer in Polish male workers in the pulp and paper industry, and 166 (95% CI:95-270) among workers engaged in paper production. METHODS: A nested case-control design within a cohort of pulp and paper workers was applied. Seventy-nine lung cancer cases and 237 "healthy" controls were selected from the cohort of 10,460 workers employed during the years 1968-1990, and observed until the end of 1995. Based on personnel files, occupational exposure was reconstructed by experts. Using a questionnaire, data on smoking habits were collected. ORs unadjusted and adjusted for smoking were calculated applying the model of conditional logistic regression. RESULTS: Occupational exposure to inorganic dusts (kaolin, lime, cement, brick, grindstone) adjusted for smoking was a significant lung cancer risk factor, with a 4.0-fold risk (95% CI:1.3-12.6), and a dose-response by cumulative dose index. Among organic dusts only wood dust increased albeit insignificantly the risk for those exposed (adjusted for smoking OR = 2.1, 95% CI:0.9-4.9), but without dose-response relationship. CONCLUSIONS: Exposure to occupational dust with relatively low content of silica, but at high concentrations may be considered as a factor increasing lung cancer risk. However, the observation made in this study should be viewed with caution as it was based on a small number of cases, and further evidence is needed to confirm or refute the authors' hypothesis.

Cardiovascular disturbances in workers exposed to carbon disulfide.

According to literature data myocardial infarction was often the first noticeable effect of CS2 on the cardiovascular system. Therefore, a question arises whether earlier circulatory lesions are absent in CS2-exposed people, or they are present but are not detectable by the diagnostic methods used. The aim of our study has been to determine whether electrocardiological disturbances do occur in exposed people, and their association, if any, with the level of CS2 exposure. Modern diagnostic methods (resting and 24-h ECG, heart rate variability, late ventricular potential analysis, and long-term blood pressure monitoring) were used to answer this question. The examination comprised 177 male workers aged 24-66 years (44+/-12), employed under conditions of the exposure for 5-38 years (14+/-9), and 93 male workers non-exposed to CS2 aged 23-65 years (41+/-12). Results of our studies show that abnormalities in resting and/or 24-h ECG recordings occur significantly more frequently in people exposed to CS2 than in the controls. ECG abnormalities were found primarily among the workers with a long, over 20-year, period of exposure. The considerable coincidence between the questionnaire and ECG results was found (87% reported complaints were confirmed by ECG data). Therefore it seems that all CS2-exposed people reporting heart symptoms should be subjected to a detailed examination. The frequency of late ventricular potentials in CS2-exposed people was not higher than in the controls. It may indicate that the ECG disturbances observed in those people were associated with functional rather than organic changes. Results of long-term ABP monitoring did not show increased incidence of elevated blood pressure in the exposed group. As the levels of the risk factors of ischemic heart disease in the exposed and control groups were similar, it seems that exposure to CS2 was responsible for the significant changes in the circulatory system.

Mortality of workers at two asbestos-cement plants in Poland.

To assess mortality rate among workers occupationally exposed to asbestos, cohort studies were carried out in two asbestos cement plants operating since the 1960s. Asbestos cement sheets for roofing and siding have been manufactured there, using mostly chrisotile, and since 1985 also crocidolite for pressure pipes. In all, the cohort comprised 3,220 workers, including 2,616 male workers. Subject to consideration were the workers employed for at least three months in the period between the onset of the production and 1980. The vital status of the subjects was traced up to 31 December 1991. The availability of the cohort was 96.8%. Workers' mortality was analysed using standardized mortality ratio (SMR). The reference group was the general population of Poland. In the male cohort, 385 cases of death were recorded. Statistically significant excess of mortality from large intestine cancer (7 cases, SMR = 264) and pleural mesothelioma (5 cases, SMR = 2846) was found. In male workers who died from pleural mesothelioma the work history ranged from 12 to 26 years. An excess mortality from pleural mesothelioma was also noted among the female workers (2 cases, SMR = 11,275). No malignant neoplasms of other locations produced significant excess mortality either in the male or female workers.

Hyposensitivity to amphetamine following exposure to chlorphenvinphos--protection by amphetamine preexposure.

We investigated the effect of an acute exposure to chlorphenvinphos (CVP), an organophosphate anticholinesterase, on amphetamine-induced open-field locomotion in rats. CVP was administered in a single i.p. dose of 1.0 mg/kg (1/10 of the LD50). All animals were challenged with 1.0 mg/kg amphetamine (AMPH) three weeks after the CVP exposure, i.e. after a time sufficient for acetylcholinesterase recovery. Some rats were also given AMPH three weeks before the CVP exposure. In rats challenged with AMPH only once after the CVP exposure, AMPH-induced open-field locomotion was significantly reduced. Such an effect was not observed in rats given AMPH three weeks before the CVP exposure. The results suggest that a single CVP exposure may result in persistent dopaminergic hyposensitivity, and that an amphetamine pretreatment may protect the rat against this effect.

Time-of-flight secondary ion mass spectrometry of matrix-diluted oligo- and polypeptides bombarded with slow and fast projectiles: positive and negative matrix and analyte ion yields, background signals, and sample aging.

Human angiotensin II, chain B of bovine insulin, and porcine insulin were determined by time-of-flight secondary ion mass spectrometry under impact of approximately 25 keV Xe+ and SF5+ ion beams and approximately 100 MeV 252Cf fission fragments. Matrix-embedded samples, dissolved in a large surplus of alpha-cyano-4-hydroxycinnamic acid, were prepared by nebulizer spray deposition, neat samples by the droplet technique. It is shown that the status of the sample can be assessed by evaluating the matrix-specific features of the mass spectra. The beneficial effect of matrix isolation was small for angiotensin but large for the insulin samples, which did not show parent peaks from neat material. Negative ion yields under SF5+ impact were up to a factor of 50 higher than with Xe+. For positive secondary ions, the enhancement was much smaller. The mass spectra produced by slow ion beams or fast fission fragments were qualitatively similar. Quantitative differences include the following: with fast projectiles the yields were about 10-30 times higher than with slow ions, but similar for negative ion emission under SF5+ bombardment; the analyte-to-matrix yield ratios were higher with slow ions and up to 250 times higher than the molar analyte concentration; for analyte ions the peak-to-background ratios were higher using slow projectiles; the fraction of carbon-rich collisionally formed molecular ions was much higher with fast projectiles. Sample aging in vacuum for up to five weeks strongly reduced the yield of protonated analyte molecules ejected by slow ion impact, but not of deprotonated species. Hence protonation seems to correlate with sample "wetness" or the presence of volatile proton-donating additives.