Heterologous Production of Antimicrobial Peptides in Propionibacterium freudenreichii

Heterologous bacteriocin production in Propionibacterium freudenreichii is described. We developed an efficient system for DNA shuttling between Escherichia coli and P. freudenreichii using vector pAMT1. It is based on the P. freudenreichii rolling-circle replicating plasmid pLME108 and carries the cml(A)/cmx(A) chloramphenicol resistance marker. Introduction of the propionicin T1 structural gene (pctA) into pAMT1 under the control of the constitutive promoter (P₄) yielded bacteriocin in amounts equal to those of the wild-type producer Propionibacterium thoenii 419. The P. freudenreichii clone showed propionicin T1 activity in coculture, killing 90% of sensitive bacteria within 48 h. The pamA gene from P. thoenii 419 encoding the protease-activated antimicrobial peptide (PAMP) was cloned and expressed in P. freudenreichii, resulting in secretion of the pro-PAMP protein. Like in the wild type, PAMP activation was dependent on externally added protease. Secretion of the antimicrobial peptide was obtained from a clone in which the pamA signal peptide and PAMP were fused in frame. The promoter region of pamA was identified by fusion of putative promoter fragments to the coding sequence of the pctA gene. The P₄ and P_pamp promoters directed constitutive gene expression, and activity of both promoters was enhanced by elements upstream of the promoter core region.     

Immunofluorescence studies on the effect of cyclic adenosine 3′,5′-monophosphate in tissue culture preparations of the cervicovaginal anlage from neonatal mice

Summary The authors have studied the production of antigenic material, specific for the cervicovaginal epithelium, in organ cultures of the cervicovaginal region from neonatal mice. In control cultures the antigenic material appeared at a time corresponding to the normal appearance under in vivo conditions. When cyclic adenosine 3, 5-monophosphate (dibutyryl derivate) was added to the medium, a strong increase of the fluorescent material was seen.This investigations was supported by grants from the Norwegian Research Council and the Norwegian Cancer Society (Landsforeningen mot Kreft).     

Intersexuality in the Pig

In 3 relatively large samples intersexuality was found in 0.2 % of the pigs. Litters with intersexes had a deficit of male piglets. The total number of piglets in such litters was correspondingly reduced. Pig losses until the age of 5 weeks were reduced in intersex litters. Other malformations, especially scrotal hernia, were more common in these litters than in normal ones. Forty-five intersexes showed big variation in the development of sexual organs. All had testicles, however. Thirteen (29 %) were so-called true hermaphrodites whereas 32 were male pseudohermaphrodites. All 42 cases being karyotyped were genetical females. There are arguments for genetical predisposition for the defect. The hereditary transmission is probably complicated. In any case it is not monogenic recessive. Practical consequences of the defect are: almost complete sterility, slaughter losses due to bad smell and taste of the meat and increased risk of cassation (local or general inflammations), increased frequency of other malformations in litters holding intersexes.     

Prophylactic iron supplementation in pregnant women in Norway

In the present study 67 non-anaemic women were randomly allocated to either 100 mg or 15 mg iron daily at about the 10. week of pregnancy. At about week 18, 30 and 36 of pregnancy, as well as 6 weeks after delivery, hemoglobin and the serum concentrations of ferritin, vitamin B12, folates, Zn, Cu and Se were monitored. Dietary allowances of other minerals and vitamins are also increased in pregnancy, and the 15 mg iron tablet was enriched with Zn (10 mg), Cu (2 mg), Se (50 microg), vitamin B12 (3 microg), and folate (0.1 mg). Neither ferritin, nor Cu, Zn or Se concentrations differed statistically significantly between the treatment groups during pregnancy. Ferritin and Zn appeared to decrease approximately parallel to the hemodilution, whereas Cu concentrations increased from a non-pregnant reference mean of 18 micromol Cu/L to a maximum mean of nearly 33 micromol Cu/L during pregnancy. Se decreased concomitantly to about 1.0 micromol Se/L. Serum folate (around 15 micromol/L) was essentially unaffected by pregnancy in the group given multivitamin/mineral supplementation, whereas the mean concentration fell below 10 micromol/L in the group supplemented with 100 mg iron daily. Our results indicate that supplementation of 15 mg Fe daily during pregnancy results in a small reduction of hemoglobin. It is suggested that additional supplementation with folate might be of importance to maintain the serum folate concentration during pregnancy.     

The influence of volume of exercise on early adaptations to strength training

The purpose of this investigation was to compare the effects of single-set strength training and 3-set strength training during the early phase of adaptation in 18 untrained male subjects (age, 20-30 years). After initial testing, subjects were randomly assigned to either the 3L-1U group (n = 8), which trained 3 sets in leg exercises and 1 set in upper-body exercises, or the 1L-3U group (n = 10), which trained 1 set in leg exercises and 3 sets in upper-body exercises. Testing was conducted at the beginning and at the end of the study and consisted of 2 maximal isometric tests (knee extension and bench press) and 6 maximal dynamic tests (1 repetition maximum [1RM] tests). Subjects trained 3 days per week for 6 weeks. After warm-up, subjects performed 3 leg exercises and 4 upper-body exercises. In both groups, each set consisted of 7 repetitions (reps) with the load supposed to induce muscular failure after the seventh rep (7RM load). After 6 weeks of training, 1RM performance in all training exercises was significantly increased (10-26%, p < 0.01) in both groups. The relative increase in 1RM load in the 3 leg exercises was significantly greater in the 3L-1U group than in the 1L-3U group (21% vs. 14%, p = 0.01). However, the relative increase in 1RM load in the 3 upper-body exercises was similar in the 3L-1U group (16%) and the 1L-3U group (14%). These results show a superior adaptation to 3-set strength training, compared with 1-set strength training, in leg exercises but not in upper-body exercises during the early phase of adaptation.     

Locomotor Activity Patterns in Ilybius fenestratus (Fabricius) (Coleoptera,Dytiscidae) in a Central Norwegian Lake,with Evidence for Breeding-Induced Arrhythmicity

The diel activity of Ilybius fenestratus was studied in Målsj?en, a lake in S?r-Tr?ndelag, Central Norway (63°14'N, 10°26'E), during 1971–1972. Adults and larvae were sampled in activity traps every week during March–October and every 2–3 weeks during November–February. The traps were emptied every 2 hrs during a 24- hr period. The diel locomotor activity of I. fenestratus was clearly nocturnal, except in the second half of July, for the males also in the first half of July, when surprisingly the population showed an arrhythmic activity pattern. The arrhythmicity is connected with the mating and egg-laying period of the species. The larvae were also nocturnal in the autumn, but showed arrhythmicity under the ice during the winter.     

Host-Specificity and Dynamics in Bacterial Communities Associated with Bloom-Forming Freshwater Phytoplankton

Many freshwater phytoplankton species have the potential to form transient nuisance blooms that affect water quality and other aquatic biota. Heterotrophic bacteria can influence such blooms via nutrient regeneration but also via antagonism and other biotic interactions. We studied the composition of bacterial communities associated with three bloom-forming freshwater phytoplankton species, the diatom Aulacoseira granulata and the cyanobacteria Microcystis aeruginosa and Cylindrospermopsis raciborskii. Experimental cultures incubated with and without lake bacteria were sampled in three different growth phases and bacterial community composition was assessed by 454-Pyrosequencing of 16S rRNA gene amplicons. Betaproteobacteria were dominant in all cultures inoculated with lake bacteria, but decreased during the experiment. In contrast, Alphaproteobacteria, which made up the second most abundant class of bacteria, increased overall during the course of the experiment. Other bacterial classes responded in contrasting ways to the experimental incubations causing significantly different bacterial communities to develop in response to host phytoplankton species, growth phase and between attached and free-living fractions. Differences in bacterial community composition between cyanobacteria and diatom cultures were greater than between the two cyanobacteria. Despite the significance, major differences between phytoplankton cultures were in the proportion of the OTUs rather than in the absence or presence of specific taxa. Different phytoplankton species favoring different bacterial communities may have important consequences for the fate of organic matter in systems where these bloom forming species occur. The dynamics and development of transient blooms may also be affected as bacterial communities seem to influence phytoplankton species growth in contrasting ways.     

Robot-Mediated Interviews - How Effective Is a Humanoid Robot as a Tool for Interviewing Young Children?

Robots have been used in a variety of education, therapy or entertainment contexts. This paper introduces the novel application of using humanoid robots for robot-mediated interviews. An experimental study examines how children’s responses towards the humanoid robot KASPAR in an interview context differ in comparison to their interaction with a human in a similar setting. Twenty-one children aged between 7 and 9 took part in this study. Each child participated in two interviews, one with an adult and one with a humanoid robot. Measures include the behavioural coding of the children’s behaviour during the interviews and questionnaire data. The questions in these interviews focused on a special event that had recently taken place in the school. The results reveal that the children interacted with KASPAR very similar to how they interacted with a human interviewer. The quantitative behaviour analysis reveal that the most notable difference between the interviews with KASPAR and the human were the duration of the interviews, the eye gaze directed towards the different interviewers, and the response time of the interviewers. These results are discussed in light of future work towards developing KASPAR as an ‘interviewer’ for young children in application areas where a robot may have advantages over a human interviewer, e.g. in police, social services, or healthcare applications.     

Complete mitochondrial genome sequences of the Arctic Ocean codfishes <Emphasis Type="Italic">Arctogadus glacialis</Emphasis> and <Emphasis Type="Italic">Boreogadus saida</Emphasis> reveal oriL and tRNA gene duplications

We have determined the complete mitochondrial genome sequences of the codfishes Arctogadus glacialis and Boreogadus saida (Order Gadiformes, Family Gadidae). The 16,644 bp and 16,745 bp mtDNAs, respectively, contain the same set of 37 structural genes found in all vertebrates analyzed so far. The gene organization is conserved compared to other Gadidae species, but with one notable exception. B. saida contains heteroplasmic rearrangement-mediated duplications that include the origin of light-strand replication and nearby tRNA genes. Examination of the mitochondrial control region of A. glacialis, B. saida, and four additional representative Gadidae genera identified a highly variable domain containing tandem repeat motifs in A. glacialis. Mitogenomic phylogeny based on the complete mitochondrial genome sequence, the concatenated protein-coding genes, and the derived protein sequences strongly supports a sister taxa affiliation of A. glacialis and B. saida.     

New Production Regulates Export Stoichiometry in the Ocean

The proportion in which carbon and growth-limiting nutrients are exported from the oceans’ productive surface layer to the deep sea is a crucial parameter in models of the biological carbon pump. Based on >400 vertical flux observations of particulate organic carbon (POC) and nitrogen (PON) from the European Arctic Ocean we show the common assumption of constant C:N stoichiometry not to be met. Exported POC:PON ratios exceeded the classical Redfield atomic ratio of 6.625 in the entire region, with the largest deviation in the deep Central Arctic Ocean. In this part the mean exported POC:PON ratio of 9.7 (a:a) implies c. 40% higher carbon export compared to Redfield-based estimates. When spatially integrated, the potential POC export in the European Arctic was 10–30% higher than suggested by calculations based on constant POC:PON ratios. We further demonstrate that the exported POC:PON ratio varies regionally in relation to nitrate-based new production over geographical scales that range from the Arctic to the subtropics, being highest in the least productive oligotrophic Central Arctic Ocean and subtropical gyres. Accounting for variations in export stoichiometry among systems of different productivity will improve the ability of models to resolve regional patterns in carbon export and, hence, the oceans’ contribution to the global carbon cycle will be predicted more accurately.