Geographical variation in egg mass and egg content in a passerine bird

Reproductive, phenotypic and life-history traits in many animal and plant taxa show geographic variation, indicating spatial variation in selection regimes. Maternal deposition to avian eggs, such as hormones, antibodies and antioxidants, critically affect development of the offspring, with long-lasting effects on the phenotype and fitness. Little is however known about large-scale geographical patterns of variation in maternal deposition to eggs. We studied geographical variation in egg components of a passerine bird, the pied flycatcher (Ficedula hypoleuca), by collecting samples from 16 populations and measuring egg and yolk mass, albumen lysozyme activity, yolk immunoglobulins, yolk androgens and yolk total carotenoids. We found significant variation among populations in most egg components, but ca. 90% of the variation was among individuals within populations. Population however explained 40% of the variation in carotenoid levels. In contrast to our hypothesis, we found geographical trends only in carotenoids, but not in any of the other egg components. Our results thus suggest high within-population variation and leave little scope for local adaptation and genetic differentiation in deposition of different egg components. The role of these maternally-derived resources in evolutionary change should be further investigated.     

Carbon source dependence and photostimulation of conidiation in Hypocrea atroviridis

Hypocrea atroviridis is frequently used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. Light is thereby believed to be the primary trigger for spore formation. In contrast, we show here that conidiation is primarily carbon source dependent and that illumination plays a catalytic role; of a total of 95 tested carbon sources, only a small set of carbohydrates, polyols, and sugar acids allowed conidiation in darkness, and on most of them, conidiation was significantly more strongly expressed in light. In addition, there are also a number of carbon sources on which H. atroviridis conidiates in darkness, but light does not further stimulate the process. Yet on another small set of carbon sources (L-sorbitol, D-fucose, D- and L-arabinose, and erythritol), H. atroviridis shows better sporulation in darkness than in light. No sporulation was observed on organic acids and amino acids. Mutants with deletions in the two blue-light receptor proteins BLR-1 and BLR-2 generally showed weaker conidiation on a smaller number of carbon sources than did the parental strain, yet they clearly sporulated on 15 and 27 of the 95 carbon sources tested, respectively. Of the carbon sources supporting sporulation, only 11 supported the conidiation of both mutants, suggesting that the BLR-1 and BLR-2 receptors are variously involved in the carbon source-dependent regulation of spore formation. The addition of cyclic AMP, which has been reported to lead to conidiation in darkness, both positively and negatively affected sporulation and resulted in different effects in the parental strain and the two Deltablr mutants. Our data show that the carbon source is the prime determinant for conidiation and that it influences the organism's regulation of conidiation by means of BLR-1 and BLR-2 and their cross talk with cyclic AMP.     

Soft sensor for monitoring biomass subpopulations in mammalian cell culture processes

Objectives

Biomass subpopulations in mammalian cell culture processes cause impurities and influence productivity, which requires this critical process parameter to be monitored in real-time.

Results

For this reason, a novel soft sensor concept for estimating viable, dead and lysed cell concentration was developed, based on the robust and cheap in situ measurements of permittivity and turbidity in combination with a simple model. It could be shown that the turbidity measurements contain information about all investigated biomass subpopulations. The novelty of the developed soft sensor is the real-time estimation of lysed cell concentration, which is directly correlated to process-related impurities such as DNA and host cell protein in the supernatant. Based on data generated by two fed-batch processes the developed soft sensor is described and discussed.

Conclusions

The presented soft sensor concept provides a tool for viable, dead and lysed cell concentration estimation in real-time with adequate accuracy and enables further applications with respect to process optimization and control.

     

Comparison of data science workflows for root cause analysis of bioprocesses

Root cause analysis (RCA) is one of the most prominent tools used to comprehensively evaluate a biopharmaceutical production process. Despite of its widespread use in industry, the Food and Drug Administration has observed a lot of unsuitable approaches for RCAs within the last years. The reasons for those unsuitable approaches are the use of incorrect variables during the analysis and the lack in process understanding, which impede correct model interpretation. Two major approaches to perform RCAs are currently dominating the chemical and pharmaceutical industry: raw data analysis and feature-based approach. Both techniques are shown to be able to identify the significant variables causing the variance of the response. Although they are different in data unfolding, the same tools as principal component analysis and partial least square regression are used in both concepts. Within this article we demonstrate the strength and weaknesses of both approaches. We proved that a fusion of both results in a comprehensive and effective workflow, which not only increases better process understanding. We demonstrate this workflow along with an example. Hence, the presented workflow allows to save analysis time and to reduce the effort of data mining by easy detection of the most important variables within the given dataset. Subsequently, the final obtained process knowledge can be translated into new hypotheses, which can be tested experimentally and thereby lead to effectively improving process robustness.

     

Hangexposition und Brutbiologie von Wasseramsel (Cinclus cinclus), Trauerschn?pper (Ficedula hypoleuca), Kohl- und Tannenmeise (Parus major, P. ater)

Zusammenfassung Im Harz besiedelten 1969–1983 Trauerschnäpper und Wasseramsel Südhänge bevorzugt und erfolgreicher als Nordhänge, Tannenmeisen dagegen Nordhänge. Siedlungsdichte, Höchstvorkommen, Gelegegröße, Zahl der flüggen Jungen pro erfolgreiches Paar, relativer Bruterfolg und Jungenproduktion (bei der Wasseramsel auch die Zahl der Zweitbruten) sind an den Süd-(Nord-) hängen größer als an den Nord-(Süd-)hängen, Legebeginn/Schlüpftermin liegen früher (Abb. 8). Zwar sind bei der Kohlmeise Gelegegröße, Zahl der flüggen Jungen pro erfolgreiches Paar und relativer Bruterfolg an den Nordhängen größer als an den Südhängen, doch die übrigen brutbiologischen Daten zeigen, daß auch für sie Südhänge nahrungsökologisch günstiger sind. Trauerschnäpper, Kohl- und Tannenmeise besiedelten bevorzugt (ca. 90 %) Hänge mit der gleichen Exposition, an der die Vögel geboren worden waren. Die unterschiedliche Bedeutung der südlichen bzw. nördlichen Hangexpositionen für die 4 Arten kann am deutlichsten aus der Zahl der flüggen Jungen pro ha oder pro km Flußlänge abgelesen werden (Abb. 8): Sie übersteigt an den Südhängen die an den Nordhängen beim Trauerschnäpper um 1000 (–2000) %, bei Wasseramsel und Kohlmeise um 65–90 %; umgekehrt ist sie bei der Tannenmeise an den Nordhängen um 100–130 % größer. Die Nahrungssituation wird als entscheidend für die Ergebnisse angesehen.

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Exposure and breeding biology ofCinclus cinclus, Ficedula hypoleuca, Parus major andP. ater

Summary From 1969 to 1983 breeding biology and settling behaviour were investigated in the Harz mountains.Ficedula hypoleuca andCinclus cinclus preferred south exposed slopes to north exposed slopes and were more successful on southern slopes. On the other handParus ater preferred northern slopes. ForFicedula hypoleuca andCinclus cinclus population density, altitudinal limit, clutch size, number of fledged young per successful pair, relative breeding success and number of fledged young per ha or per km of river bank were higher on southern than on northern slopes. The opposite was true forParus ater for northern slopes versus southern slopes (Fig. 8). InParus major clutch size, number of fledged young per successful pair and relative breeding success were also higher on northern slopes than on southern slopes. Data of breeding biology show, however, that the food ecology of south exposed slopes forParus major is also more favourable than that of north exposed slopes. For nestingFicedula hypoleuca, Parus major andP. ater prefer (to almost 90 %) the type of slope at which they had been born. The importance of north or south exposure is best shown by the number of fledged young per ha or per km of river bank (Fig. 8): On south exposed slopes it exceeds that on north exposed slopes by about 1000 (to 2000) % forFidecula hypoleuca, and by about 65–90 % forCinclus cinclus and forParus major. On the north exposed slopes, however, the number of fledged young per ha ofParus ater exceeds that of the south exposed slopes by about 100 to 130 %. The results are chiefly explained by the food situation.

     

Functional Screen of Paracrine Signals in Breast Carcinoma Fibroblasts

Stromal fibroblasts actively participate in normal mammary gland homeostasis and in breast carcinoma growth and progression by secreting paracrine factors; however, little is known about the identity of paracrine mediators in individual patients. The purpose of this study was to characterize paracrine signaling pathways between breast carcinoma cells and breast carcinoma-associated fibroblasts (CAF) or normal mammary fibroblasts (NF), respectively. CAF and NF were isolated from breast carcinoma tissue samples and adjacent normal mammary gland tissue of 28 patients. The fibroblasts were grown in 3D collagen gel co-culture with T47D human breast carcinoma cells and T47D cell growth was measured. CAF stimulated T47D cell growth to a significantly greater degree than NF. We detected a considerable inter-individual heterogeneity of paracrine interactions but identified FGF2, HB-EGF, heparanase-1 and SDF1 as factors that were consistently responsible for the activity of carcinoma-associated fibroblasts. CAF from low-grade but not high-grade carcinomas required insulin-like growth factor 1 and transforming growth factor beta 1 to stimulate carcinoma growth. Paradoxically, blocking of membrane-type 1 matrix metalloprotease stimulated T47D cell growth in co-culture with NF. The results were largely mirrored by treating the fibroblasts with siRNA oligonucleotides prior to co-culture, implicating the fibroblasts as principal production site for the secreted mediators. In summary, we identify a paracrine signaling network with inter-individual commonalities and differences. These findings have significant implications for the design of stroma-targeted therapies.     

REPRODUCTIVE COMPATIBILITY AND RDNA SEQUENCE ANALYSES IN THE SELLAPHORA PUPULA SPECIES COMPLEX (BACILLARIOPHYTA)1

We tested whether internal transcribed spacer (ITS) rDNA sequence differences are correlated with sexual compatibility in the Sellaphora pupula complex, a model system for investigations of the species concept and speciation in diatoms. The phylogenetic relationships among the demes and the systematic position of the genus within the raphid diatoms were also investigated. The division of clones of S. pupula and S. laevissima into groups, based on sequence similarities and phylogenetic analyses, resembled groupings based on sexual compatibility: A high ITS sequence divergence, making full alignment difficult or impossible, was found among clones whose gametangia do not interact, whereas there was little sequence divergence among interfertile clones. This is clearly consistent with the idea that “Z clades” exhibit less intraclade than interclade variation in ITS and, as comparisons of secondary structure models for the RECT and PSEUDOCAP clones showed, that there is an equivalence of “CBC” and Z clades in the rectangular and pseudocapitate demes of S. pupula, as earlier hypothesized for chlorophytes. Intraclonal, presumably intraindividual, variation in ITS was found in S. pupula, though with a degree of variation less than that found within a single Z clade; it was too minor to affect the interclonal relationships in the ITS phylogeny. Sellaphora, which appears monophyletic in 18S phylogenies, with Pinnularia and “Navicula”pelliculosa as its closest allies, may also include some species currently classified in Eolimna. The S. pupula–S. laevissima group began to diversify in or before the Miocene.     

Amoebae as Potential Environmental Hosts for Mycobacterium ulcerans and Other Mycobacteria, but Doubtful Actors in Buruli Ulcer Epidemiology

Background

The reservoir and mode of transmission of Mycobacterium ulcerans, the causative agent of Buruli ulcer, remain unknown. Ecological, genetic and epidemiological information nonetheless suggests that M. ulcerans may reside in aquatic protozoa.

Methodology/Principal Findings

We experimentally infected Acanthamoeba polyphaga with M. ulcerans and found that the bacilli were phagocytised, not digested and remained viable for the duration of the experiment. Furthermore, we collected 13 water, 90 biofilm and 45 detritus samples in both Buruli ulcer endemic and non-endemic communities in Ghana, from which we cultivated amoeboid protozoa and mycobacteria. M. ulcerans was not isolated, but other mycobacteria were as frequently isolated from intracellular as from extracellular sources, suggesting that they commonly infect amoebae in nature. We screened the samples as well as the amoeba cultures for the M. ulcerans markers IS2404, IS2606 and KR-B. IS2404 was detected in 2% of the environmental samples and in 4% of the amoeba cultures. The IS2404 positive amoeba cultures included up to 5 different protozoan species, and originated both from Buruli ulcer endemic and non-endemic communities.

Conclusions/Significance

This is the first report of experimental infection of amoebae with M. ulcerans and of the detection of the marker IS2404 in amoeba cultures isolated from the environment. We conclude that amoeba are potential natural hosts for M. ulcerans, yet remain sceptical about their implication in the transmission of M. ulcerans to humans and their importance in the epidemiology of Buruli ulcer.