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Biomedical subjects

R Zink

Publications and source records attributed to R Zink.

At least 19 recordsLinked to original sources
PubMed2001

Molecular microbial analysis of Bifidobacterium isolates from different environments by the species-specific amplified ribosomal DNA restriction analysis (ARDRA).

One hundred and six isolates of the genus Bifidobacterium, isolated from different environments (mainly gastrointestinal), were identified and classified taxonomically to species level by amplified ribosomal DNA restriction analysis. Two restriction endonucleases (Sau3AI and BamHI) were chosen for aligning the 16S rRNA sequences of 16 bifidobacterial species retrieved from various databases, to obtain species-specific restriction patterns. A rapid and accurate identification scheme was obtained by comparing the resulting 16S rDNA digestion profiles of 16 Bifidobacterium type-strains and 90 strains of various origins. All of the investigated strains were previously confirmed at the species level as belonging to the genus Bifidobacterium by fluorescence in-situ hybridisation and by polymerase chain reaction amplification with genus- and species-specific primers. The present work demonstrates that species-specific detection of Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium coryneforme, Bifidobacterium cuniculi, Bifidobacterium dentium, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium suis, Bifidobacterium magnum, Bifidobacterium pseudolongum, Bifidobacterium pseudocatenulatum and Bifidobacterium pullorum present in different micro-ecological environments (e.g. gastrointestinal tract) can be accomplished in a reliable, rapid and accurate manner, circumventing the recognised deficiencies of traditional identification techniques.

Journal Article↗
PubMed2001

Specific identification and targeted characterization of Bifidobacterium lactis from different environmental isolates by a combined multiplex-PCR approach.

The species Bifidobacterium lactis, with its main representative strain Bb12 (DSM 10140), is a yoghurt isolate used as a probiotic strain and is commercially applied in different types of yoghurts and infant formulas. In order to ensure the genetic identity and safety of this bacterial isolate, species- and strain-specific molecular tools for genetic fingerprinting must be available to identify isolated bifidobacteria or lactic acid bacteria from, e.g., various clinical environments of relevance in medical microbiology. Two opposing rRNA gene-targeted primers have been developed for specific detection of this microorganism by PCR. The specificity of this approach was evaluated and verified with DNA samples isolated from single and mixed cultures of bifidobacteria and lactobacilli (48 isolates, including the type strains of 29 Bifidobacterium and 9 Lactobacillus species). Furthermore, we performed a Multiplex-PCR using oligonucleotide primers targeting a specific region of the 16S rRNA gene for the genus Bifidobacterium and a conserved eubacterial 16S rDNA sequence. The specificity and sensitivity of this detection with a pure culture of B. lactis were, respectively, 100 bacteria/ml after 25 cycles of PCR and 1 to 10 bacteria/ml after a 50-cycle nested-PCR approach.

Bacterial Typing Techniques↗
PubMed2000

Basic features of the stress response in three species of bifidobacteria: B. longum, B. adolescentis, and B. breve.

The presence of the dnaK heat shock gene could be demonstrated for B. longum NCC481, B. longum NCC490, B. longum NCC585, B. adolescentis NCC251, and B. breve NCC298. Induction of dnaK on the transcriptional level was shown for NCC251 and NCC481 by increasing temperatures. NCC251 showed an additional chaperone-induction after salt or bile-salt treatment. In both strains preconditioning with bile-salts protected against otherwise lethal concentrations thereof. NCC251 when subjected to a heat stress was able to survive an otherwise lethal temperature (55 degrees C). Cross-protection was demonstrated for NCC251 since salt pretreatment resulted in increased tolerance after freeze-thawing cycles or lethal heat stress.

Bifidobacterium↗
PubMed2000

Genomics, molecular genetics and the food industry.

The production of foods for an increasingly informed and selective consumer requires the coordinated activities of the various branches of the food chain in order to provide convenient, wholesome, tasty, safe and affordable foods. Also, the size and complexity of the food sector ensures that no single player can control a single process from seed production, through farming and processing to a final product marketed in a retail outlet. Furthermore, the scientific advances in genome research and their exploitation via biotechnology is leading to a technology driven revolution that will have advantages for the consumer and food industry alike. The segment of food processing aids, namely industrial enzymes which have been enhanced by the use of biotechnology, has proven invaluable in the production of enzymes with greater purity and flexibility while ensuring a sustainable and cheap supply. Such enzymes produced in safe GRAS microorganisms are available today and are being used in the production of foods. A second rapidly evolving segment that is already having an impact on our foods may be found in the new genetically modified crops. While the most notorious examples today were developed by the seed companies for the agro-industry directed at the farming sector for cost saving production of the main agronomical products like soya and maize, its benefits are also being seen in the reduced use of herbicides and pesticides which will have long term benefits for the environment. Technology-driven advances for the food processing industry and the consumer are being developed and may be divided into two separate sectors that will be presented in greater detail: 1. The application of genome research and biotechnology to the breeding and development of improved plants. This may be as an aid for the cataloging of industrially important plant varieties, the rapid identification of key quality traits for enhanced classical breeding programs, or the genetic modification of important plants for improved processing properties or health characteristics. 2. The development of advanced microorganisms for food fermentations with improved flavor production, health or technological characteristics. Both yeasts and bacteria have been developed that fulfill these requirements, but are as yet not used in the production of foods.

Bacteriophages↗
PubMed2000

Iron requirement of Lactobacillus spp. in completely chemically defined growth media.

A completely chemically-defined growth medium, containing guanine, thymine, cytidine, 2'-deoxyadenosine and 2'-deoxyuridine as DNA precursors, was developed for Lactobacillus johnsonii, on the basis of statistically designed techniques suitable for other lactobacilli. Particular focus was given to the nucleotide composition of different defined media, and to the specific nucleotide requirements of Lact. johnsonii. Most of the lactobacilli tested grew in a medium containing five free bases, four ribonucleosides or five deoxyribonucleosides. Adenine and guanine were replaceable by inosine. The requirement for thymine and cytosine was satisfied with uracil. The presence of inosine and uracil was identified as being essential for the growth of different Lactobacillus species, displaying their inability to synthesize purines and pyrimidines de novo. Defined recipes with different nucleotide composition were used to investigate iron requirements of lactobacilli. Only marginal differences in growth were observed in iron-depleted media supplemented with five free bases, four ribonucleosides or five deoxyribonucleosides; iron depletion had a greater effect on growth when inosine and uracil were supplied as the only nucleotide sources. The results suggest that iron plays a role in the pyrimidine and purine metabolism of lactobacilli. Lactobacillus spp., particularly Lact. johnsonii, require iron under particular environmental conditions with limited or specific nucleotide sources.

Culture Media↗
PubMed1999

Galvanic stimulation in bilateral vestibular failure: 3-D ocular motor effects.

Bilateral galvanic vestibular stimulation (GVS) with current intensity of 3 mA was applied at mastoid level in 11 patients with chronic bilateral vestibular failure, in order to determine ocular motor responses by 3-D video-oculography. The following abnormal features were found: (1) a predominantly torsional or mixed torsional-horizontal nystagmus at the onset of stimulation with lower current intensities (1.0-3.0 mA) in nine patients; (2) a reduced amplitude of tonic ocular torsion by about 50% in nine patients (1.3 +/- 0.6 degrees at 3 mA); (3) a nystagmus in the opposite direction at stimulation offset in five patients (rebound); (4) no eye movements at all in a patient with bilateral nerve failure. GVS stimulates the vestibular nerve, thus allowing differentiation of nerve failure from labyrinthine failure. The low thresholds for initiating nystagmus and the rebound, which appear to be the most typical features of bilateral labyrinthine failure, can be explained by central compensation mechanisms.

Acceleration↗
PubMed1998

Cerebral functional magnetic resonance imaging of vestibular, auditory, and nociceptive areas during galvanic stimulation.

Cerebral activation was investigated with functional magnetic resonance imaging (fMRI) during galvanic stimulation of the mastoid in 6 normal volunteers. Cutaneous stimulation at the neck C4-5 level served as a control. During mastoid stimulation, bilateral vestibular activation occurred in the posterior insula (parietoinsular vestibular cortex, PIVC), the transverse temporal (Heschl's) gyrus, and thalamic pulvinar. The cutaneous pain elicited by galvanic stimulation caused bilateral activity of the medial part of the insula and the anterior median thalamus. Thus, galvanic stimulation at the mastoid level activates cortical areas of three different sensory systems in the insulathalamic region, the vestibular, the auditory, and the nociceptive systems.

Adult↗
PubMed1998

Effects of galvanic vestibular stimulation on otolithic and semicircular canal eye movements and perceived vertical.

OBJECTIVE: The aim of this study was to determine the otolithic and semicircular canal effects of galvanic vestibular stimulation with increasing current strengths on eye movements and the perception of verticality. METHODS: We measured (1) 3-dimensional eye movements, (2) subjective tilt of the peripheral visual field, and (3) subjective tilt of a central vertical line in 12 healthy subjects during galvanic vestibular stimulation. A rectangular, unipolar binaural electric current was applied to each subject's mastoid. RESULTS: Anodal stimulation of the right mastoid led to an ipsiversive tonic ocular torsion of up to 5.4 degrees, to a contralateral tilt of both the peripheral visual field (1-9 degrees), and a central vertical line (0.5-6.2 degrees) increasing in amplitude with increasing current strengths applied. This reflects otolith stimulation. In most subjects, current strengths of 3 mA or more elicited a slight (horizontal-) torsional nystagmus (amplitude 1-2 degrees) that was superimposed on static torsion. This reflects horizontal and vertical semicircular canal stimulation. A correlation was found in the amount of the 3 measured parameters and the strength of the applied current. CONCLUSIONS: Thus, galvanic vestibular stimulation at low current intensities (1-3 mA) preferably excites otolith responses, which increase with increasing current intensity. With higher current intensity above 3 mA, additional semicircular canal responses are elicited in the form of horizontal-rotatory nystagmus superimposed on static torsional deviations. The lack of a vertical deviation and nystagmus can be explained by the counterdirected vertical components of the anterior and posterior semicircular canal.

Adult↗
PubMed1997

Galvanic vestibular stimulation in humans: effects on otolith function in roll.

The effects of unilateral galvanic vestibular stimulation on (1) ocular torsion, (2) subjective tilt of the peripheral visual field, and (3) subjective tilt of a foveal vertical line were measured in 12 healthy subjects. A rectangular, unipolar binaural electric current was applied to the subject' s mastoid. Anodal stimulation of the right mastoid led to an ipsiversive tonic ocular torsion (0.5-3.7 degrees) and to a contralateral tilt of both the peripheral visual field (1-9 degrees), and a foveal vertical line (0.5-6.2 degrees). There was a correlation between the amount of the three measured parameters and the strength of the applied current. Static ocular torsion, central and peripheral visual tilts represent stimulus-induced tonic otolith imbalance between the two labyrinths. Thus, galvanic vestibular stimulation not only affects dynamic semicircular canal input but also static otolith input in the roll plane.

Adult↗
PubMed1996

[Cost reduction in long-term nursing thanks to a memory clinic--results of a case control study].

Between 1991 and 1995, 334 persons from the city of Zurich have been investigated at the gerontologic counselling facility of the memory clinic of the nursing home Entlisberg near Zurich. 49 (14.7%) of these persons suffered from a treatable disease (16 from depression, 13 from chronic intoxication, 20 from other treatable diseases), 10 from other disturbances (debility or postapoplectic state), and 275 from dementia. In 45% of the cases dementia was moderate, in 47% moderate to severe, and in 8% severe. In most cases dementia was of Alzheimer's type (60.7%), others were of multi-infarct type (17.5%), of mixed type (13.1%), or other types (8.7%). Until march 1996, 64 (19.4%) of these 334 on the average 74-year-old persons treated in the memory clinic have been admitted to a city nursing home. They have compared to a random sample, including 312 demented persons of similar ages and similar date of admittance to the same institutions. The proportion of demented persons cared before admittance by a spouse was about 28% in both groups. On an average, the mini mental state (Zurich variant) at admittance to a nursing home was 10.95 points in patients from the memory clinic, compared to 13.43 points in the control group. Thus, patients that have formerly been advised and trained entered the nursing home in a state of dementia more advanced by 2.48 points. Based on an average yearly demelioration by about 2.4 points, this means admittance to a nursing home is delayed by 376 days. Patients for whom admittance to a nursing home could be avoided completely by the treatment proposed by the counselling were not considered in this calculation.

Aged↗
PubMed1996

Determinants of a normal (versus impaired) oral glucose tolerance after combined pancreas-kidney transplantation in IDDM patients.

After successful pancreas transplantation, insulin-dependent diabetic patients are characterized by a normal or at worst impaired oral glucose tolerance (World Health Organisation criteria). It is not known which pathophysiological mechanisms cause the difference between normal and impaired oral glucose tolerance. Therefore, we studied 41 patients after successful combined pancreas-kidney transplantation using stimulation in the fasting state with oral glucose (75 g), intravenous glucose (0.33 g/kg) and glucagon bolus injection (1 mg i.v.). Glucose (glucose oxidase), insulin and C-peptide (immunoassay) were measured. Repeated-measures analysis of variance and multiple regression analysis were used to analyse the results which showed: 28 patients had a normal, and 13 patients had an impaired oral glucose tolerance. Impaired oral glucose tolerance was associated with a greatly reduced early phase insulin secretory response (insulin p < 0.0001; C-peptide p = 0.037). Age (p = 0.65), body mass index (p = 0.94), immunosuppressive therapy (cyclosporin A p = 0.84; predniso(lo)ne p = 0.91; azathioprine p = 0.60) and additional clinical parameters were not different. Reduced insulin secretory responses in patients with impaired oral glucose tolerance were also found with intravenous glucose or glucagon stimulations. Exocrine secretion (alpha-amylase in 24-h urine collections) also demonstrated reduced pancreatic function in these patients (-46%; p = 0.04). Multiple regression analysis showed a significant correlation of 120-min glucose with ischaemia time (p = 0.003) and the number of HLA-DR mismatches (p = 0.026), but not with HLA-AB-mismatches (p = 0.084). In conclusion, the pathophysiological basis of impaired oral glucose tolerance after pancreas transplantation is a reduced insulin secretory capacity. Transplant damage is most likely caused by perioperative influences (ischaemia) and by the extent of rejection damage related, for example, to DR-mis-matches.

Adult↗
PubMed1995

Characterization of cryptic prophages (monocins) in Listeria and sequence analysis of a holin/endolysin gene.

Monocins in Listeria were induced by UV-irradiation of liquid cultures, and defective phage particles were purified from the lysates. Electron microscopy showed flexible, non-contractile bacteriophage-tail-like particles, consisting of specific proteins of molecular mass 20-45 kDa and pI 4.6-6.7. These particles were able to lyse listerial cells. DNA sequence homologies between chromosomal DNA of monocin-producing strains and labelled Listeria phage DNAs were inferred from DNA/DNA hybridizations, suggesting that most of the prophage DNA is still present in the listerial chromosome. An endolysin gene cpl2438 was cloned from listerial chromosomal DNA and was identified by its expression of lytic activity against Listeria cells in a bioassay. The gene consists of 864 nt encoding a protein of 287 aa with a calculated molecular mass of 32975 Da (CPL2438). This is in good agreement with the size of a protein observed in SDS-PAGE after overexpression of the lytic protein in Escherichia coli. The nucleotide sequence of a putative holin gene (hol2438, 291 nt) upstream of cpl2438 was determined after PCR-amplification of listerial DNA and it shows typical features common to the holin gene family. Expression of the encoded protein (HOL2438, 95 aa, 10.1 kDa) in E. coli was found to be lethal for the host cells. The results underline the close relationship between monocins and intact Listeria bacteriophages, indicating that monocins are incompletely assembled phage particles derived from cryptic prophages of Listeria, probably including the phage lysin.

Amino Acid Sequence↗
PubMed1994

Supplementary Listeria-typing with defective Listeria phage particles (monocins).

The potential use of monocins for listeria typing was investigated. Monocins are defective phage particles still capable of lysing listerial cells. They were induced by u.v.-irradiation, precipitated with polyethylene glycol and purified by density gradient centrifugation. Using 26 monocins, it was possible to type 48% of Listeria monocytogenes strains, 92% of L. innocua strains and 94% of L. ivanovii strains. Overall typability of 480 strains was 68%. None of the monocins was able to lyse L. grayi. Monocin typing was found to be a valuable supplementary tool for typing strains which were non-typable by the Weihenstephan phage typing set. A combination of phage typing and monocin typing increased overall typability of Listeria strains to 95%.

Bacteriophage Typing↗
PubMed1994

Taxonomical classification of 20 newly isolated Listeria bacteriophages by electron microscopy and protein analysis.

A set of 20 newly isolated temperate bacteriophages for phage typing of the pathogen Listeria monocytogenes has been investigated by means of electron microscopy and electrophoretic analysis of phage structural proteins. All phages had isometric capsids (60-64 nm diameter), and long contractile or non-contractile tails of 170-320 nm length. They could be classified into 2 morphotypes (A1 and B1) and were assigned to 3 listeriaphage species (4211, 2671, and 2389). Individual protein profiles were generated by SDS-PAGE of viral polypeptides, as well as isoelectric focusing of solubilized phage proteins in immobilized pH gradient gels. The major structural proteins ranged in size from approximately 15 to 38 kD, and showed isoelectric points from pI 4.3 to 6.2. Protein compositions permitted the differentiation of individual phages as well as the recognition and grouping of similar viruses.

Bacteriophage Typing↗
PubMed1993

Isolation, classification and molecular characterization of bacteriophages for Enterobacter species.

Out of 22 Enterobacter phages investigated, nine were found to be suitable for phage typing based on their different lytic spectra on 398 strains of Enterobacter spp. isolated from milk powder and other foods. These phages were compared on the basis of morphology, protein composition, restriction endonuclease patterns and DNA-DNA hybridization. Two phages (WS-EP19, WS-EP13) belonged to the Podoviridae family (morphotype C1), and three (WS-EP20, WS-EP26, WS-EP28) were classified as Siphoviridae (morphotype B1). The other four phages were Myoviridae of the morphological groups A1 (WS-EP57) and A2 (WS-EP32, WS-EP94, WS-EP96). SDS-PAGE revealed individual protein profiles for each phage, which corresponded to different restriction enzyme fragment patterns. DNA-DNA hybridization demonstrated the close relationship of phages WS-EP20 and WS-EP26, and of WS-EP94 and WS-EP96. In general, a good correlation was found between groupings obtained with the various methods. The nine phages could be attributed to existing enterobacterial phage species although some differences to the described type phages were observed.

Bacteriophage Typing↗
PubMed1992

Classification of virulent and temperate bacteriophages of Listeria spp. on the basis of morphology and protein analysis.

A set of 22 phages of Listeria species (listeriaphages) (21 temperate and 1 virulent) were compared on the basis of morphology and protein composition. All 22 phages had icosahedral heads and exhibited either contractile or noncontractile tails. They represented two different morphotypes. Twenty phages belonged to the Siphoviridae family and could be differentiated only on the basis of tail length. Accordingly, they could be assigned to previously defined listeriaphage species. Two other phages were classified as members of the Myoviridae family, one of which (A511) should be regarded as a new species. It was found to be substantially different from all other known listeriaphages. All phages exhibited typical protein profiles, which were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequent laser densitometrical analysis of the gels. It was then possible to distinguish eight protein subgroups on the basis of unique protein patterns. This classification corresponds well to the previous groupings based on host range. Most of the phages revealed two or three major proteins ranging from 21 to 24 kDa and 30 to 36 kDa. In addition, at least 10 minor proteins could be observed for each phage. Our results indicate that the major proteins are structural proteins of the capsid and tail, and the protein band ranging from 30 to 35 kDa could clearly be assigned to the proteins of the phage capsid.

Bacteriophages↗