Intersectional strategy to study cortical inhibitory parvalbumin-expressing interneurons.

Parvalbumin-expressing (PV) interneurons are key neuronal elements to a global excitatory-inhibitory balance in normal cortical functioning. To better understand the circuit functions of PV interneurons, reliable animal models are needed. This study investigated the sensitivity and specificity of the most frequently used PV-Cre/tdTomato mouse line in this regard.

Targeted RNA next generation sequencing analysis of cervical smears can predict the presence of hrHPV-induced cervical lesions.

Background: Because most cervical cancers are caused by high-risk human papillomaviruses (hrHPVs), cervical cancer prevention programs increasingly employ hrHPV testing as a primary test. The high sensitivity of HPV tests is accompanied by low specificity, resulting in high rates of overdiagnosis and overtreatment. Targeted circular probe-based RNA next generation sequencing (ciRNAseq) allows for the quantitative detection of RNAs of interest with high sequencing depth.

Novel high-resolution targeted sequencing of the cervicovaginal microbiome.

Background: The cervicovaginal microbiome (CVM) plays a significant role in women's cervical health and disease. Microbial alterations at the species level and characteristic community state types (CST) have been associated with acquisition and persistence of high-risk human papillomavirus (hrHPV) infections that may result in progression of cervical lesions to malignancy. Current sequencing methods, especially most commonly used multiplex 16S rRNA gene sequencing, struggle to fully clarify these changes because they generally fail to provide sufficient taxonomic resolution to adequately perform species-level associative studies.

Are the metabolic components of crassulacean acid metabolism up-regulated in response to an increase in oxidative burden?

In the halophytic species Mesembryanthemum crystallinum, crassulacean acid metabolism (CAM) may be induced by a range of abiotic factors including drought, salinity, high light intensity, low temperature, and anoxia. A key biotic consequence of all these environmental changes is the generation of reactive oxygen species in planta that can elicit potentially damaging oxidative reactions and/or act as signals for engaging mechanisms that alleviate oxidative stress. However, induction of CAM per se also has the potential for increasing the oxidative burden via the enhanced internal O2 concentrations that develop behind closed stomata during daytime decarboxylation.

Differences in the activities of some antioxidant enzymes and in H2O2 content during rhizogenesis and somatic embryogenesis in callus cultures of the ice plant.

Callus was obtained from hypocotyls of Mesembryanthemum crystallinum seedlings cultured on two types of medium-germination medium (GM) and callus induction medium (CIM). Following subculture on shoot induction medium SIM1, the callus formed on CIM medium regenerated roots or somatic embryos, while that obtained on GM medium was non-regenerative. The activities of CuZn-superoxidase dismutase (SOD) were comparable in all calli, but the activities of FeSOD and MnSOD varied according to the activity of photosystem II and the regenerative potential of the tissues.

Malate accumulation in different organs of Mesembryanthemum crystallinum L. following age-dependent or salinity-triggered CAM metabolism.

Different organs of Mesembryanthemum crystallinum exhibit differing levels of CAM (Crassulacean acid metabolism), identifiable by quantification of nocturnal malate accumulation. Shoots and also basal parts of young leaves were observed to accumulate high concentrations of malate. It was typically found in mature leaves and especially prominent in plants subjected to salt stress.

14-3-3 isoforms and pattern formation during barley microspore embryogenesis.

The members of the 14-3-3 isoform family have been shown to be developmentally regulated during animal embryogenesis, where they take part in cell differentiation processes. 14-3-3 isoform-specific expression patterns were studied in plant embryogenic processes, using barley (Hordeum vulgare L.) microspore embryogenesis as a model system.

Insertion of pea lectin into a phospholipid monolayer.

Pea lectin (PSL) is a secretory sugar-binding protein, readily soluble in aqueous solutions of low osmolarity. However, PSL also appears to be associated with the plasma membrane at the tip of young pea root hairs. By using the Wilhelmy plate method, we found that PSL can insert into a lipid monolayer.

Sugar-binding activity of pea (Pisum sativum) lectin is essential for heterologous infection of transgenic white clover hairy roots by Rhizobium leguminosarum biovar viciae.

Legume lectin stimulates infection of roots in the symbiosis between leguminous plants and bacteria of the genus Rhizobium. Introduction of the Pisum sativum lectin gene (psl) into white clover hairy roots enables heterologous infection and nodulation by the pea symbiont R. leguminosarum biovar viciae (R.

Expression of the plum pox coat protein gene in transgenic Nicotiana tabacum plants.

Plant expression vector pBI 121 containing the gene encoding coat protein of Plum Pox Virus of the Skierniewice isolate (CP PPV-S) was prepared (clone pCM1). The construct was used for transformation of Nicotiana tabacum plants using an Agrobacterium tumefaciens based system. About 82% of kanamycin resistant plant lines contained a transgene (the sequence of CP PPV-S) but only 81% of them actively expressed the PPV-S coat protein gene as measured by RT-PCR.

Mutational analysis of the sugar-binding site of pea lectin.

Comparison of x-ray crystal structures of several legume lectins, co-crystallized with sugar molecules, showed a strong conservation of amino acid residues directly involved in ligand binding. For pea (Pisum sativum) lectin (PSL), these conserved amino acids can be classified into three groups: (I) D81 and N125, present in all legume lectins studied so far; (II) G99 and G216, conserved in almost all legume lectins; and (III) A217 and E218, which are only found in Vicieae lectins and are possibly determinants of sugar-binding specificity. Each of these amino acids in PSL was changed by site-directed mutagenesis, resulting in PSL molecules with single substitutions: for group I D81A, D81N, N125A; for group II G99R, G216L; and for group III A217L, E218Q, respectively.

Pea (Pisum sativum L.) seed isolectins 1 and 2 and pea root lectin result from carboxypeptidase-like processing of a single gene product.

The complete amino acid sequences of the alpha-subunits of pea (Pisum sativum L.) seed and root lectin, the C-terminal amino acids of the beta-subunits of pea seed lectin, and most of the sequence of the beta-subunit of pea root lectin were determined. In contrast to earlier reports it was shown that the beta-subunits of both seed isolectins end at Asn-181.

Destabilization of pea lectin by substitution of a single amino acid in a surface loop.

Legume lectins are considered to be antinutritional factors (ANF) in the animal feeding industry. Inactivation of ANF is an important element in processing of food. In our study on the stability of Pisum sativum L.

Mutational analysis of pea lectin. Substitution of Asn125 for Asp in the monosaccharide-binding site eliminates mannose/glucose-binding activity.

As part of a strategy to determine the precise role of pea (Pisum sativum) lectin, Psl, in nodulation of pea by Rhizobium leguminosarum, mutations were introduced into the genetic determinant for pea lectin by site-directed mutagenesis using PCR. Introduction of a specific mutation, N125D, into a central area of the sugar-binding site resulted in complete loss of binding of Psl to dextran as well as of mannose/glucose-sensitive haemagglutination activity. As a control, substitution of an adjacent residue, A126V, did not have any detectable influence on sugar-binding activity.

The promoter of the rice gene GOS2 is active in various different monocot tissues and binds rice nuclear factor ASF-1.

A single copy gene has been isolated, termed GOS2, from rice. Sequence comparison revealed highly similar genes in mammals and yeast, indicating that GOS2 encodes an evolutionary conserved protein. GOS2 mRNA was detected in all tissues examined.

Structure and expression of a root-specific rice gene.

Two rice cDNA clones (COS6 and COS9) were isolated, corresponding to genes that were highly expressed in roots from seedlings and mature plants. A genomic clone (GOS9) corresponding to cDNA clone COS9 was isolated and the intron/exon structure was determined by comparing the nucleotide sequences of the mRNA and the genomic clone. 5' ends and 3' ends of the mRNA were determined by primer extension and S1-nuclease mapping respectively.

[Hypersensitivity to alcoholic beverages during treatment with ketoconazole].

A 41-year-old female patient is described who developed intolerance to alcoholic beverages while using ketoconazole, 200 mg daily. Two similar cases have previously been described in the literature. The frequency and the pharmacology of this suspected interaction are unknown.

Effects of mutations in the TATA box region of the Agrobacterium T-cyt gene on its transcription in plant tissues.

We have generated mutations in the promoter region of the octopine type cytokinin gene of Agrobacterium tumefaciens, and studied their effects on mRNA formation in different plant species. The promoter region of this gene contains several putative TATA boxes. Phenotypic expression and Northern blot hybridization showed that TATA boxes are essential for expression, but that one TATA box leads to wild-type transcript levels.

Plant expression signals of the Agrobacterium T-cyt gene.

Within the 5' and 3' non-coding regions of the T-cyt gene from the octopine T-DNA of Agrobacterium tumefaciens sequences required for expression of this gene in plant cells were identified by deletion mutagenesis. The results show that 184 bp of the 5' non-coding region and 270 bp of the 3' non-coding region are sufficient for wild-type expression. Within the 5' non-coding region two essential expression signals were identified: (1.

The limited host range of an Agrobacterium tumefaciens strain extended by a cytokinin gene from a wide host range T-region.

The host range of Agrobacterium tumefaciens strain LBA649 (pTiAg57) is limited to grapevine and a few other plant species. Its host range was extended through the introduction of the T-region from the wide host range octopine plasmid pTiAch5. In contrast, R prime plasmids harboring the entire wide host range virulence region were unable to achieve this effect.

Intrauterine growth retardation: a study of long-term morbidity.

Reported is the second phase of a prospective follow-up study of 76 growth-retarded children who were mature at birth and a control group of 88 children who had weights appropriate for gestational age at birth. Follow-up assessments of motor, cognitive, and language development were made between 1 and 6 years of age. The children of the intrauterine growth retardation (IUGR) group continued to be smaller than the children of the control group between 12 and 60 months of age.

A Collaborative Study of the Delvotest-P Method to Detect Low Concentrations of Penicillin in Milk.

During a collaborative study at a large number of milk-receiving stations from a dairy industry in the Netherlands, in which penicillin was added in various concentrations of 0, 4, 5, 6, 7, 8 and 10 IU/1 to milk samples, the Delvotest-P method gave reliable results in almost all instances. Of the 22 milk samples that were tested, both undiluted and dilluted 1:1, by up to 17 control stations in seven experiments, a total of 518 readings were obtained. There were no negative readings among the 249 samples that contained 4 or more IU penicillin per liter and only one doubtful reading among 186 samples that were free of penicillin.