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J Harder

Publications and source records attributed to J Harder.

At least 37 records · Page 2Linked to original sources

Anaerobic mineralization of quaternary carbon atoms: isolation of denitrifying bacteria on dimethylmalonate.

The microbial capacity to degrade simple organic compounds with quaternary carbon atoms was demonstrated by enrichment and isolation of five denitrifying strains on dimethylmalonate as the sole electron donor and carbon source. Quantitative growth experiments showed a complete mineralization of dimethylmalonate. According to phylogenetic analysis of the complete 16S rRNA genes, two strains isolated from activated sewage sludge were related to the genus Paracoccus within the alpha-Proteobacteria (98.0 and 98.2% 16S rRNA gene similarity to Paracoccus denitrificans(T)), and three strains isolated from freshwater ditches were affiliated with the beta-Proteobacteria (97.4 and 98.3% 16S rRNA gene similarity to Herbaspirillum seropedicae(T) and Acidovorax facilis(T), respectively). Most-probable-number determinations for denitrifying populations in sewage sludge yielded 4.6 x 10(4) dimethylmalonate-utilizing cells ml(-1), representing up to 0.4% of the total culturable nitrate-reducing population.

Anaerobiosis↗

Community size and metabolic rates of psychrophilic sulfate-reducing bacteria in Arctic marine sediments.

The numbers of sulfate reducers in two Arctic sediments with in situ temperatures of 2.6 and -1.7 degrees C were determined. Most-probable-number counts were higher at 10 degrees C than at 20 degrees C, indicating the predominance of a psychrophilic community. Mean specific sulfate reduction rates of 19 isolated psychrophiles were compared to corresponding rates of 9 marine, mesophilic sulfate-reducing bacteria. The results indicate that, as a physiological adaptation to the permanently cold Arctic environment, psychrophilic sulfate reducers have considerably higher specific metabolic rates than their mesophilic counterparts at similarly low temperatures.

Arctic Regions↗

Differential mucin MUC7 gene expression in invasive bladder carcinoma in contrast to uniform MUC1 and MUC2 gene expression in both normal urothelium and bladder carcinoma.

Mucins (MUCs) are high molecular weight membrane glycoproteins. The gene expression of MUCs (MUC1-MUC8) may change characteristically during malignant transformation of epithelial tissues. Total RNA was isolated from the four bladder cancer cell lines RT4, 647V, HT1376, and 486P (pathological gradings between G1 and G4) and 17 samples of transitional cell carcinomas, as well as 16 samples of normal human urothelium of the bladder from surgically removed specimens. The RNA samples were studied with MUC1-, MUC2- and MUC7-specific nested reverse transcription-PCRs. Gene expression of MUC1 and MUC2 was found positive in all normal, as well as in malignant, tissue samples and in the tumor cell lines. In contrast, gene expression of MUC7 was only detected in bladder cancer cell lines and samples of invasive transitional cell carcinomas, but neither in superficial, noninvasive bladder tumors nor normal bladder urothelium. Only one of the samples of normal urothelium obtained from 16 different tumor-bearing bladders was positive for MUC7 gene expression. These results suggest a differential MUC7 gene expression with the onset of malignant transformation of the bladder urothelium.

Carcinoma, Transitional Cell↗

Antileukoprotease in human skin: an antibiotic peptide constitutively produced by keratinocytes.

Antileukoprotease (ALP), also known as mucous protease inhibitor or secretory leukoprotease inhibitor, resembles one of the major antiproteases present in human body fluids. It is capable of preventing proteolytic degradation of extracellular matrix proteins by neutrophil-derived serine proteases. ALP was isolated from human callus and detected in supernatants of cultured human primary keratinocytes. ALP mRNA was constitutively expressed in keratinocytes and the expression was not significantly affected by TNF alpha or Interferon gamma stimulation. In microbicidal assays recombinant ALP exhibited antimicrobial activity against several human skin associated microorganisms like P. aeruginosa, S. aureus, S. epidermidis, and C. albicans, indicating that ALP may actively participate in mechanisms allowing homeostasis of bacterial and yeast colonization on human skin. Thus, ALP represents a major soluble serine protease inhibitor and antimicrobial agent expressed in human skin and seems to contribute to the high resistance of the epidermis against proteolysis and infections.

Anti-Bacterial Agents↗

Factors affecting the prognosis of Ebstein's anomaly during fetal life.

BACKGROUND: The echocardiographic criteria that have been used to evaluate severity of Ebstein's anomaly in utero are the same as those applied after birth. OBJECTIVE: The objective of this study was to establish prognostic criteria that take into account the peculiarities of the fetal hemodynamics. METHOD: The video recordings of eight fetuses with Ebstein's anomaly were retrospectively reviewed. RESULTS: The following indexes had no prognostic significance either on fetal or neonatal outcome: the ratio of functional tricuspid opening over the diameter of the annulus, the degree of displacement of the tricuspid valve opening, and the degree of tricuspid regurgitation. The index of severity (based on the surfaces of right atrium + atrialized right ventricle) and the cardiothoracic ratio had a significant impact only on neonatal survival. The smallest fossa ovalis were found in two fetuses who had hydrops. Fetuses who reached term without problems had higher left ventricular outputs. A positive linear correlation was found between the z score of the left ventricular output and the size of the fossa ovalis (r = 0.81, p < 0.05). CONCLUSION: The prognosis of Ebstein's anomaly during fetal life is not influenced by criteria described for postnatal life and may be related to factors that control the volume load of the left ventricle.

Ebstein Anomaly↗

Alcaligenes defragrans sp. nov., description of four strains isolated on alkenoic monoterpenes ((+)-menthene, alpha-pinene, 2-carene, and alpha-phellandrene) and nitrate.

Four pseudomonad strains 51Men, 54Pin, 62Car and 65Phen were recently isolated on the monoterpenes (+)-menthene, alpha-pinene, 2-carene and alpha-phellandrene as sole carbon source and nitrate as electron acceptor. These bacteria were characterised. The motile, mesophilic, Gram-negative rods had a strictly respiratory metabolism. Monoterpenes as carbon sources were completely mineralised to carbon dioxide. The physiology of all strains was very similar, but displayed an individual utilisation preference for the isolation substrate. The fatty acid composition of whole cells showed a high degree of similarity to that of Alcaligenes faecalis. Comparative 16S rDNA data analysis placed the isolates into the beta-subclass of Proteobacteria in a common offshoot together with Alcaligenes and Bordetella species. On the basis of these characteristics, the strains are described as a new species belonging to the genus Alcaligenes, A. defragrans sp. nov., with strain 54Pin (DSM 12141T) as type strain.

Alcaligenes↗

Thauera linaloolentis sp. nov. and Thauera terpenica sp. nov., isolated on oxygen-containing monoterpenes (linalool, menthol, and eucalyptol) nitrate.

The monoterpenes menthol, linalool, and eucalyptol were recently used as sole electron donor and carbon source for the isolation of three denitrifying bacterial strains 21Mol, 47Lol, and 58Eu. The motile, mesophilic, Gram-negative rods had a strictly respiratory metabolism. Monoterpenes were completely mineralised to carbon dioxide, nitrate was reduced to dinitrogen. Strain 47Lol utilised aliphatic monoterpenes, strain 21Mol oxygenated monocyclic monoterpenes, and strain 58Eu the bicyclic eucalyptol and monocyclic monoterpene alkenes. The fatty acid composition of the strains indicated an allocation to the rRNA group III of pseudomonads. Comparative 16S rRNA gene sequence analyses revealed that the new isolates can be assigned as members of the genus Thauera within the beta subclass of Proteobacteria. DNA-DNA hybridisation studies indicated a relateness of 68.5% between strains 21Mol and 58Eu which shared 36.0% and 40.6% DNA similarity with strain 47Lol. The strains are described as new species belonging to the genus Thauera, strain 47Lol (DSM 12138T) as T. linaloolentis sp. nov. and strains 21Mol and 58Eu as T. terpenica sp. nov. with strain 58Eu (DSM 12139T) as type strain.

Acyclic Monoterpenes↗

Biotransformation of monoterpenes, bile acids, and other isoprenoids in anaerobic ecosystems.

Isoprenoic compounds play a major part in the global carbon cycle. Biosynthesis and mineralization by aerobic bacteria have been intensively studied. This review describes our knowledge on the anaerobic metabolism of isoprenoids, mainly by denitrifying and fermentative bacteria. Nitrate-reducing beta-Proteobacteria were isolated on monoterpenes as sole carbon source and electron donor. Thauera spp. were obtained on the oxygen-containing monoterpenes linalool, menthol, and eucalyptol. Several strains of Alcaligenes defragrans were isolated on unsaturated monoterpenes as growth substrates. A novel denitrifying beta-Proteobacterium, strain 72Chol, mineralizes cholesterol completely to carbon dioxide. Physiological studies showed the presence of several oxidative pathways in these microorganisms. Investigations by organic geochemists indicate possible contributions of anaerobes to early diagenetic processes. One example, the formation of p-cymene from monoterpenes, could indeed be detected in methanogenic enrichment cultures. In man, cholic acid (CA) and chenodeoxycholic acid (CDCA), are synthesized in the liver from cholesterol. During their enterohepatic circulation, bile acids are biotransformed by the intestinal microflora into a variety of metabolites. Known bacterial biotranformations of conjugated bile acids include: deconjugation, oxidation of hydroxy groups at C-3, C-7 and C-12 with formation of oxo bile acids and reduction of these oxo groups to either alpha- or beta-configuration. Quantitatively, the most important bacterial biotransformation is the 7 alpha-dehydroxylation of CA and CDCA yielding deoxycholic acid and lithocholic acid, respectively. The 7 alpha-dehydroxylation of CA occurs via a novel six-step biochemical pathway. The genes encoding several enzymes that either transport bile acids or catalyze various reactions in the 7 alpha-dehydroxylation pathway of Eubacterium sp. strain VPI 12708 have been cloned, expressed in Escherichia coli, purified, and characterized.

Bacteria, Anaerobic↗

[Environmental medical action required on exposure to noise].

Basing on request for an expertise on the medical effects of noise on healthy adults, permissible values for mainly traffic noise were estimated from the literature. If the permissible values are exceeded, preventive medical action is necessary. Below these values the probability of noise-induced health hazards is essentially zero. The authors presume that noise levels above these permissible values, but below the limit of bearability, are an increasing danger to health. Preventive medical action must be weighed against the risk involved in persistent noise, depending on the situation and the need protection. The effect of noise as a health hazard is, apart from the mechanical damage to the inner ear, a psycho-physiological deregulation which can be either indirectly due to the annoyance or directly caused by stress of the vegetative-hormonal system. Therefore different permissible values for annoyance, the stress on the vegetative-hormonal system and for the loss of hearing are suggested, for both continual and maximum noise levels. In addition, de-regulation depends on the time of acoustic exposure because the sensitivity to noise follows a 24-hour cycle (circadian rhythm). It is therefore necessary to determine personal permissible limits for the nocturnal noise (sleep disorder). Permissible values for children are also suggested.

Adolescent↗

Mapping of the gene encoding human beta-defensin-2 (DEFB2) to chromosome region 8p22-p23.1.

We recently reported the isolation of human beta-defensin-2 (hBD-2), a novel epithelia-derived peptide antibiotic belonging to the beta-defensin family. hBD-2 is expressed in skin and epithelia of the airway system, where it is believed to contribute to its antimicrobial defense. By fluorescence in situ hybridization using a hBD-2 genomic DNA probe and subsequent fluorescence R-banding, the hBD-2 gene (HGMW-approved symbol DEFB2) was assigned to human chromosome region 8p22-p23.1. PCR with a set of CEPH YAC clones spanning this chromosomal region revealed CEPH YACs 773G4, 920D12, and 820B4 to contain the hBD-2 gene. Relying on the preexisting physical maps of 8p22-p23.1, the hBD-2 gene was mapped in close proximity to D8S1993 (WI-9956) within the interval flanked by D8S552 and D8S1130 (CHLC.GATA25C10). The fact that all currently described genes encoding defensins map to chromosome 8p21-pter suggests that a gene cluster in this chromosomal region may play a major role in antimicrobial defense.

Blood Proteins↗

Anaerobic mineralization of cholesterol by a novel type of denitrifying bacterium

A novel denitrifying bacterium, strain 72Chol, was enriched and isolated under strictly anoxic conditions on cholesterol as sole electron donor and carbon source. Strain 72Chol grew on cholesterol with oxygen or nitrate as electron acceptor. Strictly anaerobic growth in the absence of oxygen was demonstrated using chemically reduced culture media. During anaerobic growth, nitrate was initially reduced to nitrite. At low nitrate concentrations, nitrite was further reduced to nitrogen gas. Ammonia was assimilated. The degradation balance measured in cholesterol-limited cultures and the amounts of carbon dioxide, nitrite, and nitrogen gas formed during the microbial process indicated a complete oxidation of cholesterol to carbon dioxide. A phylogenetic comparison based on total 16S rDNA sequence analysis indicated that the isolated micro-organism, strain 72Chol, belongs to the beta2-subgroup in the Proteobacteria and is related to Rhodocyclus, Thauera, and Azoarcus species.

Journal Article↗

Evolution of fetal ventricular aneurysms and diverticula of the heart: an echocardiographic study.

Due to the rarity of congenital ventricular diverticula and aneurysms, their natural history remains unclear. An excellent prognosis has been suggested for those cases diagnosed during fetal life: From October 1992 to January 1996 seven fetuses were diagnosed with ventricular diverticula or aneurysms. Gestational age ranged from 18 to 36 weeks. The indications for fetal echocardiogram were cardiomegaly, abnormal four-chamber view, a large pericardial effusion, and hydrothorax. Echocardiography revealed a moderate sized apical left ventricular aneurysm (2), a small subvalvular right ventricular diverticulum (1), small apical right ventricular diverticulum (2), a large submitral left ventricular aneurysm (1), and a large diverticulum arising from the lateral free wall of the left ventricle (1). Decreased left ventricular function was detected in three fetuses with left ventricular aneurysms. Two fetuses with large lesions, developed hydrops and died in utero. Postnatal echocardiograms confirmed prenatal findings in all survivors. All infants remained asymptomatic, with age on follow-up from 8 to 24 months. An accurate diagnosis of ventricular diverticula and aneurysms is feasible prenatally. Outcome depends on the size and progression of the lesion. The presence of a large pericardial effusion in three cases with a diverticula was noted.

Diagnosis, Differential↗

Urinary secretory immunoglobulin A and free secretory component in pyelonephritis.

The immune defense mechanisms of mucosal surfaces involve secretory immunoglobulin A (sIgA) antibodies and, to a lesser degree, other specific and nonspecific immune factors. These antibodies are dependent on a secretory component (SC) for their transmission through the epithelium. This SC is also secreted without Ig as free SC (FSC). The kidney does produce these proteins; however, the ability of the lower urinary tract to secrete them has not been shown. Thus, an upper urinary tract infection should produce more urinary sIg and possibly more FSC than a lower tract infection. To demonstrate this, urine was obtained from normal controls (N = 33), cystitis patients (N = 22), and pyelonephritis patients (N = 27). Monoclonal antibodies binding to specific conformational epitopes were used in an enzyme-linked immunosorbent assay to detect the levels of sIgA and FSC in these groups. Previous sIgA measurements have been hampered by lack of specificity of the capture antibody. Urine creatinine was obtained to correct for the effect of diuresis. A one-tailed Student's t-test for nonparametric populations was performed to assess differences. The sIgA levels in the normal and cystitis groups were equivalent (1.4 micrograms/mg/mL and 1.3 micrograms/mg/mL, respectively; P = 0.32). When these two groups were compared with the pyelonephritis group (24.1 micrograms/mg/mL), a statistically significant difference was seen (P = 0.012 and P = 0.011, respectively), with no overlap. There was a statistical difference in the levels of FSC in these same groups, but a large degree of overlap.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Microbial degradation of monoterpenes in the absence of molecular oxygen.

Anaerobic degradation of natural monoterpenes by microorganisms was evaluated by using Pseudomonas citronellolis DSM 50332 and enrichment cultures containing nitrate as an electron acceptor. P. citronellolis grew anaerobically on 3,7-dimethyl-1-octanol and citronellol but not on geraniol, nerol, and alicyclic monoterpenes. In contrast, several a-, mono-, and bicyclic monoterpenes supported microbial growth and denitrification in enrichment cultures. We found that consumption of linalool, menthol, menth-1-ene, alpha-phellandrene, limonene, 2-carene, alpha-pinene, and fenchone in enrichment cultures depended on the presence of living microorganisms and nitrate. In these experiments, the ratios of number of electrons derived from complete substrate oxidation to number of electrons derived from nitrate reduction ranged from 1.2:1 to 2.9:1. Microbial degradation was accompanied by the formation of small traces of monoterpenes, which were characterized by gas chromatography-mass spectroscopy. The formation of geraniol and geranial from linalool suggested that a 3,1-hydroxyl-delta 1-delta 2-mutase reaction initiates linalool degradation. Seven strains of motile, oval to rod-shaped, facultatively denitrifying bacteria were isolated on agar bottle plates by using linalool, menthol, menth-1-ene, alpha-phellandrene, 2-carene, eucalyptol, and alpha-pinene as sole carbon and energy sources.

Anaerobiosis↗

A possible glycine radical in anaerobic ribonucleotide reductase from Escherichia coli: nucleotide sequence of the cloned nrdD gene.

During anaerobic growth of Escherichia coli an oxygen-sensitive ribonucleoside-triphosphate reductase, different from the aerobic ribonucleoside diphosphate-reductase (EC 1.17.4.1), produces the deoxyribonucleoside triphosphates required for DNA replication. The gene for the anaerobic enzyme has now been cloned and was found to contain a 2136-nucleotide coding region, corresponding to 712 amino acid residues, and an Fnr binding site 228 base pairs upstream of the initiator ATG. The deduced amino acid sequence shows 72% identity to a gene of coliphage T4, sunY, hitherto of unknown function, suggesting that the virus codes for its own anaerobic reductase. The location of an organic free radical formed during activation of the bacterial anaerobic reductase is proposed to be on Gly-681, since the pentapeptide RVCGY at positions 678-682 shows a striking similarity to the C-terminal sequence. RVSGY, of pyruvate formate-lyase. During activation of the anaerobically induced pyruvate formate-lyase, the glycine residue of the pentapeptide becomes an organic radical [Wagner, A. F. V., Frey, M., Neugebauer, F. A., Schäfer, W. & Knappe, J. (1992) Proc. Natl. Acad. Sci. USA 89, 996-1000]. The gene for the anaerobic reductase is located at a position around 96 min on the E. coli genomic map.

Amino Acid Sequence↗

Ribonucleotide reductases and their occurrence in microorganisms: a link to the RNA/DNA transition.

The evolution of a deoxyribonucleotide synthesizing ribonucleotide reductase might have initiated the transition from the ancient RNA world into the prevailing DNA world. At least five classes of ribonucleotide reductases have evolved. The ancient enzyme has not been identified. A reconstruction of the first ribonucleotide reductase requires knowledge of contemporary enzymes and of microbial evolution. Experimental work on the former focuses on few organisms, whereas the latter is now well understood on the basis of ribosomal RNA sequences. Deoxyribonucleotide formation has not been investigated in many evolutionary important microorganisms. This review covers our knowledge on deoxyribonucleotide synthesis in microorganisms and the distribution of ribonucleotide reductases in nature. Ecological constraints on enzyme evolution and knowledge deficiencies emerge from complete coverage of the phylogenetic groups.

Animals↗

Characterization of components of the anaerobic ribonucleotide reductase system from Escherichia coli.

Anaerobic growth of Escherichia coli induces an oxygen-sensitive ribonucleoside triphosphate reductase system, different from the aerobic ribonucleoside diphosphate reductase (EC 1.17.4.1) of aerobic E. coli and higher organisms (Fontecave, M., Eliasson, R., and Reichard, P. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 2147-2151). We have now purified and characterized two proteins from the anaerobic system, provisionally named dA1 and dA3. dA3 is the actual ribonucleoside triphosphate reductase; dA1 has an auxiliary function. From gel filtration, dA1 and dA3 have apparent molecular masses of 27 and 145 kDa, respectively. In denaturing gel electrophoresis, dA3 gives two bands of closely related polypeptides with apparent molecular masses of 77 (beta 1) and 74 (beta 2) kDa. Immunological and structural evidence suggests that beta 2 is a degradation product of beta 1 and that the active enzyme is a dimer of beta 1. dA1 activity coincides on denaturing gels with a band of 29 kDa and thus appears to be a monomer. The reaction requires, in addition, an extract from E. coli heated for 30 min at 100 degrees C. Potassium is one required component, but one or several others remain unidentified and are provisionally designated fraction RT. With dA3, dA1, RT, and potassium ions, CTP reduction shows absolute requirements for S-adenosylmethionine, NADPH (with NADH as a less active substitute), dithiothreitol, and magnesium ions, and is strongly stimulated by ATP, probably acting as an allosteric effector. Micromolar concentrations of several chelators inhibit CTP reduction completely, suggesting the involvement of (a) transition metal(s).

Aerobiosis↗