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K Valentin

Publications and source records attributed to K Valentin.

At least 19 recordsLinked to original sources

Picoeukaryotic plankton diversity at the Helgoland time series site as assessed by three molecular methods.

We analyzed picoeukaryote assemblages in the German Bight at the Helgoland time series site by sequencing cloned eukaryotic 18S rRNA genes in six genetic libraries plus one library from the Orkney Islands from a cruise of opportunity. The libraries were constructed from environmental samples collected at different periods of the year. The same samples were also analyzed using a fingerprinting technique, single-strand conformational polymorphism (SSCP), and DNA microarrays with class-level oligonucleotide probes. One hundred unique clones were analyzed from each library, thus insuring over 85% coverage of the library. The V4 region of the 18S rRNA gene was sequenced from each of these clones, thus providing the most discrimination among the clones. The nonphotosynthetic picoeukaryotic component dominated over the photosynthetic one and was represented by the ciliates at 45% and group II alveolates at 42%. Prasinophytes dominated the photosynthetic group at 40%, but other picoplankton groups, such as bolidomonads and chrysophytes, were also present. Totally novel groups were found in the cryptomonads and in the dinoflagellates. A new algal group sister to the cryptophyte nuclear gene and the glaucocystophytes was also found. These three groups have been found in other picoeukaryotic planktonic clone libraries. SSCP analyses at closer time intervals suggest that clone libraries should be made at weekly intervals if succession in the picoeukaryotic plankton community is to be monitored accurately. A comparison of annual samples suggests thatthere appears to be an annual cycle with regard to species composition. Microarray analysis supported the clone library data and offered a faster means of community analysis, which can be performed with similar accuracy and with higher throughput for a more in-depth analysis.

Animals↗

The structure and gene repertoire of an ancient red algal plastid genome.

Photosynthetic eukaryotes can, according to features of their chloroplasts, be divided into two major groups: the red and the green lineage of plastid evolution. To extend the knowledge about the evolution of the red lineage we have sequenced and analyzed the chloroplast genome (cp-genome) of Cyanidium caldarium RK1, a unicellular red alga (AF022186). The analysis revealed that this genome shows several unusual structural features, such as a hypothetical hairpin structure in a gene-free region and absence of large repeat units. We provide evidence that this structural organization of the cp-genome of C. caldarium may be that of the most ancient cp-genome so far described. We also compared the cp-genome of C. caldarium to the other known cp-genomes of the red lineage. The cp-genome of C. caldarium cannot be readily aligned with that of Porphyra purpurea, a multicellular red alga, or Guillardia theta due to a displacement of a region of the cp-genome. The phylogenetic tree reveals that the secondary endosymbiosis, through which G. theta evolved, took place after the separation of the ancestors of C. caldarium and P. purpurea. We found several genes unique to the cp-genome of C. caldarium. Five of them seem to be involved in the building of bacterial cell envelopes and may be responsible for the thermotolerance of the chloroplast of this alga. Two additional genes may play a role in stabilizing the photosynthetic machinery against salt stress and detoxification of the chloroplast. Thus, these genes may be unique to the cp-genome of C. caldarium and may be required for the endurance of the extreme living conditions of this alga.

Adaptation, Physiological↗

Phylogeny and expression of the secA gene from a chromophytic alga--implications for the evolution of plastids and sec-dependent protein translocation.

In bacteria many periplasmatic proteins are exported via the sec-dependent pathway. A homologous apparatus was found to be involved in the transport of proteins across the thylakoid membrane in plastids. In the present study additional data on the phylogeny and expression of one of the genes essential in this process, secA, is presented. For the first time, transcriptional activity of secA in the plastid was detected. When secA is used as a phylogenetic marker for plastid evolution it demonstrates a large phylogenetic distance between chlorophytic and non-chlorophytic (i.e. rhodophytic) primary plastids. This distance could be explained by assuming polyphyly for major plastid lineages. Moreover, it was found that two types of secA genes may exist in plastids. Whether or not these are involved in different protein translocation processes is presently unknown. In an attempt to identify further candidates, i.e. non-photosynthesis-related proteins, for sec-dependent protein transport, an SbpA protein was detected in chromophytic plastids by the use of a peptide antibody.

Algal Proteins↗

A model for the evolution of the plastid sec apparatus inferred from secY gene phylogeny.

Plastids possess a bacteria-like sec apparatus that is involved in protein import into the thylakoid lumen. We have analyzed one of the genes essential for this process, secY. A secY gene from the unicellular red alga Cyanidium caldarium was found to be transcriptionally active, demonstrating for the first time that secY is functional in a plastid. Unlike the situation seen in bacteria the C. caldarium gene is transcribed monocistronically, despite the fact that it is part of a large ribosomal gene cluster that resembles bacterial spc operons. A molecular phylogeny is presented for 8 plastid-encoded secY genes, four of which have not been published yet. In this analysis plastid secY genes fall into two classes. One of these, comprising of genes from multicellular red algae and Cryptophyta, clusters in a neighbour-joining tree with a cyanobacterial counterpart. Separated from the aforesaid are secY genes from Chromophyta, Glaucocystophyta and a unicellular red alga. All plastid and cyanobacterial sequences are located on the same branch, separated from bacterial homologues. We postulate that the two classes of secY genes are paralogous, i.e. their gene products are involved in different protein translocation processes. Based on this assumption a model for the evolution of the plastid sec apparatus is presented.

Cloning, Molecular↗

Glutamate synthase is plastid-encoded in a red alga: implications for the evolution of glutamate synthases.

An actively transcribed gene (glsF) encoding for ferredoxin-dependent glutamate synthase (Fd-GOGAT) was found on the plastid genome of the multicellular red alga Antithamnion sp. Fd-GOGAT is not plastid-encoded in chlorophytic plants, demonstrating that red algal plastid genomes encode for additional functions when compared to those known from green chloroplasts. Moreover, our results suggest that the plant Fd-GOGAT has an endosymbiotic origin. The same may not be true for NADPH-dependent GOGAT. In Antithamnion glsF is flanked upstream by cpcBA and downstream by psaC and is transcribed monocistronically. Implications of these results for the evolution of GOGAT enzymes and the plastid genome are discussed.

Amino Acid Sequence↗

SecA is plastid-encoded in a red alga: implications for the evolution of plastid genomes and the thylakoid protein import apparatus.

Partial sequence analysis of the plastid DNA (ptDNA) from a red alga, Antithamnion sp., revealed the presence of a homologue to the Escherichia coli secA gene as well as two open reading frames (ORF 510, ORF 179). In addition a sec Y homologue has been detected on the plastid genome by heterologous hybridization. None of these genes has been found in completely sequenced chlorophytic plastid genomes. SecA and secY gene copies were also detected in the ptDNA of a chromophytic alga, indicating that secA Y may be ubiquitous in rhodophytes and chromophytes. The significance of these findings for the evolution of plastid genomes and the thylakoid protein import mechanism is discussed.

Adenosine Triphosphatases↗

Organization and expression of a phycobiliprotein gene cluster from the unicellular red alga Cyanidium caldarium.

We have sequenced a plastid gene cluster from the unicellular red alga Cyanidium caldarium which is located downstream from the psbA gene and contains, in the following order, genes for a beta-allophycocyanin-like protein (apcB'), a putative 9.5 kDa allophycocyanin linker protein (apcL9.5) and a putative 29 kDa phycocyanin linker protein (cpcL29). The apcB' and apcL9.5 genes are organized in the form of an operon. The cpcL29 gene is transcribed monocistronically from the opposite strand of DNA. Both transcription units are probably terminated at a 25 bp inverted repeat 3 and 5 bp downstream of the stop codons of the apcL9.5 and cpcL29 genes, respectively. The levels of both transcripts are greatly reduced in the dark as is the psbA transcript. Downstream from the phycobiliprotein gene cluster two open reading frames (ORFs) were found which are homologous to ORFs from plastid DNAs and cyanelle DNA of Cyanophora paradoxa. Sequence homologies between genes analysed in this study and corresponding genes from cyanobacteria, chlorophytic plastids and cyanelles point to a large phylogenetic distance between the plastids of Cyanidium and cyanobacteria and other plastid types.

Amino Acid Sequence↗

Organization and sequence of photosynthetic genes from the plastid genome of the holoparasitic flowering plant Cuscuta reflexa.

We have cloned and sequenced an area of about 6 kb of the plastid DNA (ptDNA) from the holoparasitic plant Cuscuta reflexa. This region contains (in the following order) genes for the cytochrome b6 f-complex subunit V (petG), tRNA(Val) (trnV), tRNA(Met) (trnM), the epsilon- and beta-subunit of the chloroplast ATP-synthase (atpE and atpB) and the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; rbcL). In addition we identified other photosynthesis-related genes (atpA, petB, psaA, psbA, psbB, psbC, and psbD) in C. reflexa by heterologous hybridization. The gene arrangement of the sequenced area is, except for the petG gene, the same as in ptDNAs of other higher plants (e.g. Nicotiana tabacum). Sequence homologies between the Cuscuta genes and corresponding genes from higher plants are in the range of 90%. The only significant difference is that the rbcL gene of C. reflexa encodes a polypeptide which is 18-23 amino acids longer than in other higher plants. This is remarkable since C. reflexa has lost its ability to grow photoautotrophically. The transcript level of the rbcL gene, however, is strongly reduced as compared to tobacco. These findings are compatible with results from Western blotting analysis, where no Rubisco large subunit was detectable, and with the lack of Rubisco activity in crude extracts of C. reflexa.

Amino Acid Sequence↗

Structural similarities between psbA genes from red and brown algae.

The single copy psbA genes from the multicellular red alga Antithamnion spec. and the brown alga Ectocarpus siliculosus have been cloned and sequenced and monocistronic transcripts have been detected. Both genes contain an insertion of 21 bp at the 3' end which was also found in cyanobacteria and which is absent in chloroplasts and the chlorophyll b-containing prochlorophyte Prochlorothrix hollandica. These findings are in agreement with the hypothesis of a polyphyletic origin of plastids. Plastids of red and brown algae appear to be closely related.

Amino Acid Sequence↗

Rubisco genes indicate a close phylogenetic relation between the plastids of Chromophyta and Rhodophyta.

The genes for both subunits of Rubisco (rbcL, rbcS) are located on the plastome of the brown alga Ectocarpus siliculosus (Chromophyta, Phaeophyceae). The organization of these genes in the form of an operon was similar to that found in rhodoplasts, cyanobacteria and the plastids of Cryptomonas phi. Sequence analysis of the complete operon revealed a high degree of homology and great structural similarities to corresponding genes from two red algae. In contrast, sequence homology to Rubisco genes from chloroplasts and cyanobacteria was much lower. This clearly indicated a close phylogenetic relationship between the plastids of Rhodophyta and Chromophyta which seem to have evolved independently from the chloroplasts (polyphyletic origin). Our data suggest that the plastids of Chromophyta and Cryptophyta have originated from endosymbiotic unicellular red algae. Surprisingly, red and brown algal Rubiscos show a significantly higher degree of homology to that from a hydrogen bacterium than to those from cyanobacteria.

Amino Acid Sequence↗

Structure of the rubisco operon from the multicellular red alga Antithamnion spec.

In the multicellular red alga Antithamnion spec. both rubisco genes (rbcL and rbcS) are encoded on the plastid DNA (ptDNA). Both genes are separated by a short A/T-rich spacer of 100 bp and are cotranscribed into an mRNA of approximately 2.7 kb. These findings are in extensive agreement with those obtained from two unicellular red algae (Porphyridium aerugineum and Cyanidium caldarium). The large subunit (LSU) of rubisco shows an amino acid homology of 82-87% with the LSUs from the two unicellular red algae and only about 55% to LSUs from green algae, higher plants and two cyanobacteria. The small subunit (SSU) of rubisco is more similar to those from the unicellular red algae and two algae which are members of the Chromophyta (about 60% homology) than to cyanobacterial and higher plant proteins (27-36% homology). These data indicate that rhodoplasts originated independently from the chloroplast line. The plastids of chromophytes and rhodophytes appear to be closely related.

Amino Acid Sequence↗

The psbA-gene from a red alga resembles those from cyanobacteria and cyanelles.

Plastid DNA (ptDNA) from the unicellular red alga Cyanidium caldarium was isolated. A 5.8 kb Eco RI, fragment containing the entire psbA-gene was cloned and the nucleotide sequence of the psbA-gene determined. At the carboxyl terminus the encoded protein (D1) contains the seven amino acid-insertion which was found to be typical of the cyanobacteria and the cyanelles of Cyanophora paradoxa. However, the overall sequence homology does not support a direct relationship between the plastids of Cyanidium, cyanelles and the cyanobacteria. As in other photosynthetic organisms the psbA-gene is transcribed as a monocistronic mRNA. The ribosomal RNA operon was located 4 kb upstream of the psbA-gene.

Amino Acid Sequence↗

Nucleotide sequence of the gene for the large subunit of Rubisco from Cyanophora paradoxa--phylogenetic implications.

The gene (rbcL) for the large subunit (LSU) of Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Cyanophora paradoxa was cloned and the nucleotide sequence determined. Sequence homologies to rbcL genes from other sources clearly indicated a close phylogenetic relationship between the photosynthetic organelles of Cyanophora (cyanelles), green chloroplasts and cyanobacteria. Our data support the hypothesis that the cyanelles of Cyanophora may represent a closely related, but independent, side line to chloroplast evolution. Cyanelles and rhodoplasts or phaeoplasts seem not to be related.

Amino Acid Sequence↗

Structure of the Rubisco operon from the unicellular red alga Cyanidium caldarium: evidence for a polyphyletic origin of the plastids.

The genes for both subunits of ribulose-1,5-bisphosphate-carboxylase/oxygenase (Rubisco) were located on the plastid DNA (ptDNA) of the unicellular red alga Cyanidium caldarium. Both genes are organized together in an operon. The sequence homology of both genes to the corresponding genes from the unicellular red alga Porphyridium aerugineum is remarkably high, whereas homology to Rubisco genes from chloroplasts and two recent cyanobacteria is significantly lower. These data provide strong evidence for a polyphyletic origin of chloroplasts and rhodoplasts. In addition the genes for the small subunit of Rubisco (rbcS) from red algae show about 60% homology to rbcS genes from cryptophytes and chromophytes. Thus, homologies in the rbcS gene indicate a close phylogenetic relationship between rhodoplasts and the plastids of Chromophyta.

Amino Acid Sequence↗

The genes of both subunits of ribulose-1,5-bisphosphate carboxylase constitute an operon on the plastome of a red alga.

Plastid (pt) DNA from the red alga Porphyridium aerugineum was purified by CsCl gradient centrifugation. An EcoRI library of the ptDNA was screened with a gene probe specific for the gene encoding the large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco EC 4.1.1.39) from spinach. A 5.8 kb EcoRI clone containing the LSU gene (rbcL) was isolated and the DNA sequence of the Porphyridium rbcL gene and its flanking regions was determined. An open reading frame was found 130 bp downstream from the rbcL gene that shows homology to genes coding for the small subunit of Rubisco (rbcS) from higher plants and cyanobacteria. Both genes (rbcL + rbcS) are cotranscribed. Comparison of rbcL and rbcS sequences from Porphyridium, higher plants and cyanobacteria seems to reveal a remarkable evolutionary distance between the plastids of the red algae (rhodoplasts), chloroplasts and cyanobacteria.

Amino Acid Sequence↗

Primary bronchial and ovarian carcinoid tumors in the same patient: a case report and histopathologic analysis.

The first case report of a 58-year-old woman with two different primary carcinoid tumors is presented. The bronchial carcinoid exhibited a mosaic pattern, whereas the ovarian tumor showed both insular and trabecular patterns. No teratomatous component was observed in the ovary. Both tumors presented an argyrophil reaction and neuron-specific enolase (NSE) immunoreactivity. The ovarian carcinoid contained a cell population of argentaffin and serotonin immunoreactive cells, while the bronchial carcinoid was unreactive. Peptide hormone immunostaining disclosed the presence of glucagon and pancreatic polypeptide (PP) in the bronchial carcinoid, whereas the ovarian carcinoid showed no immunoreactivity with the antisera used. The patient was treated surgically (bilobectomy of the right lung, 1978, and total abdominal hysterectomy and bilateral salpingo-oophorectomy, 1984) and is now well with normal blood chemistry, normal urinary HIAA, and normal routine X rays of the lungs and the gastrointestinal tract, as well as normal ultrasonography of the abdomen and the liver.

Bronchial Neoplasms↗

Home testing for pregnancy--can it be recommended?

A kit for home testing for pregnancy (Predictor) was tested by 201 volunteers with a menstrual delay of eight days or more who had requested a pregnancy test at a pharmacy. This number corresponds to two-thirds of the women attending the pharmacy for this purpose during the study period. The same urine sample was also tested by experienced analysts with Pregnosticon All-In--an essentially identical method with a guaranteed sensitivity of 1000 U/1 (2nd IS)--as well as assayed for choriogonadotrophin (CG) content by a sensitive and specific radioimmunoassay. Of 101 samples with a CG concentration of more than 1000 U/1 all were positive with the home test. Of 95 samples with a CG concentration below 1000 U/1, 91 were recorded as negative, two as equivocal and two as positive. The figures for sensitivity and predictive values of positive and negative test results were thus 98-100%, and those for Pregnosticon All-In were 99-100%. We conclude that home testing for pregnancy with suitable reagents may offer acceptable sensitivity and efficiency.

Chorionic Gonadotropin↗