Chechenian Parents: How to Improve the Mental Health of a Child Following War and Flight

Bakgrunn: Mange barn og unge med flyktningbakgrunn sliter med psykiske vansker. Dette er en utfordring for helsetjenesten. Hensikt: Å beskrive tsjetsjenske foreldres opplevelse av foreldrerollen i henholdsvis Tsjetsjenia og Norge, og hvilke tiltak de opplever som best for deres barns psykiske helse. Metode: Deskriptiv studie med en kvalitativ tilnærming. Ti foreldre er intervjuet. Data er analysert med innholdsanalyse. Temaene i intervjuguiden: Foreldrerollen i hjemlandet. Barnas psykiske situasjon. Foreldrenes ønsker, synspunkter og tanker om hva som kan bedre evt. dårlig psykisk helse hos barnet. Hvordan introduksjonsordningen påvirker foreldrerollen. Funn: Foreldrene forteller at mange av barna/ungdommene, etter mange år i Norge, har det dårlig psykisk og sosialt. De er ensomme, og har problemer med å finne seg til rette blant venner og i fritidsaktiviteter. Noen har atferdsforstyrrelser, og sosial mestring og funksjon er vanskelig. Barnas dårlige psykiske og sosiale situasjon gjør dem ekstra krevende for foreldrene. Informanter beskriver manglende erfaring med ansvar for og oppdragelse av barn.  Ved flyttingen til Norge har de reist fra oppdragerkompetansen i familie og nettverk. Samtidig forteller de om store forskjeller i mål og verdier i Norge og hjemlandet. Særlig mødrene forteller om en meget slitsom hverdag, som mor, deltager i introduksjonsordningen  og med egne psykiske vansker. Tidsmarginene i det daglige blir knappe, og i mange tilfeller umulige, og dette gir en høy stressfaktor. Foreldrene ønsker hjelp til helsefremmende tiltak som fritidsaktiviteter, kulturaktiviteter, hjelp til å etablere nettverk, mestring og foreldreveiledning, men har dårlig erfaring med terapi til barna. Aller mest ønsker de tid til å være foreldre for barna sine. Konklusjoner: Mottak av store familier med traumatiske opplevelser hos foreldre og barn er komplisert, og krever et samordnet tilbud i kommunen, hvor barneperspektivet må være like mye i fokus som foreldrenes kvalifisering, gjerne med en egen introduksjonslov for barna. Spesielt må det i større grad tilrettelegges for god foreldrefunksjonBackground: Many refugee children struggle with mental problems. This presents a challenge for the Norwegian health service. Purpose: This thesis aims to describe Chechenian parents’ experience of parenting in Chechnya and Norway respectively, and which initiatives they perceive to be best for their children’s mental health. Method: This descriptive study used a qualitative approach to interview ten parents who had immigrated to Norway from Chechnya. The themes in the interview guide were:  Parenting role in the homeland; the children’s psychological situation; the wishes, viewpoints and thoughts of the parents concerning what might improve the possible poor mental health of the child; how the rigours of participation in the qualification programme demanded by immigration laws affect the parenting role. Data were analysed by content analysis. Findings: The parents reported that, after many years in Norway, their children and teenagers experience both mental and social difficulties including loneliness, difficulty fitting in with friends and adjusting to leisure activities.  Some are behaviourally disturbed, and functioning socially is difficult. The children’s poor mental health and social challenges place great demands on the parents. Informants explained that they lack experience in having responsibility for raising children. Moving to Norway, they have left behind the child-rearing skills found in their family and social network. At the same time, they describe considerable differences in the parental goals and values of their homeland and Norway. Mothers especially, described a very exhausting daily life whilst participating in the qualification programme, balancing parental responsibilities whilst having their own mental difficulties. Having little or no free time increased their stress levels. Parents want help with health-promoting initiatives such as leisure activities, network building, cultural activities, and teaching of parenting skills, but their experience of child-therapy has been negative. Most of all, they want time to be parents for their children. Conclusions: The reception of large families where both parents and children have endured traumatic experiences is complicated, and demands a coordinated effort from the local council, where the child’s perspective must be as central as the parents’ circumstances, preferably with a separate qualification programme for children. In particular, programmes must be organised with good parenting function in mind.ISBN 978-91-85721-63-4</p

Interspecific interactions of heterotrophic bacteria during chitin degradation

In their natural habitats, bacteria live in multi-species microbial communities and are, thus, constantly interacting with bacteria of other phylogenetic groups. In order to prevail in these interspecific interactions, such as the competition for nutrients, bacteria have developed numerous strategies. During the degradation of polymers such interspecific interactions are likely to occur, because degradation starts as an extracellular process. In one possible interaction scenario, investor bacteria, which invest energy in the production of extracellular enzymes, face the danger of being exploited by opportunistic bacteria that compete for degradation products. To investigate such a scenario and to characterize the strategies employed by the bacteria involved, we established two co-culture model-systems consisting of bacteria that co-exist in aquatic environments and with the polymer chitin as carbon, nitrogen, and energy source. Aeromonas hydrophila strain AH-1N, which releases extracellular chitinases, was employed as investor bacterium in both co-cultures.In the first co-culture, chitin embedded in agarose served as substrate. Flavobacterium sp. strain 4D9, which cannot degrade embedded chitin due to its cell-associated chitinases, was employed as opportunistic bacterium. The strategies applied by strain 4D9 in order to acquire nutrients included active integration into the biofilm formed by strain AH-1N on the chitin beads and interception of the chitin monomer N-acetylglucosamine (GlcNAc), leading to overgrowth of strain AH-1N by strain 4D9 in the biofilm.In the second co-culture, suspended chitin served as substrate. Pseudomonas aeruginosa strain PAO1, which is unable to degrade chitin, was employed as opportunistic bacterium. In the first phase of the co-culture, strain PAO1 grew with ammonium, acetate, and possibly GlcNAc and other compounds, which were released by strain AH-1N. In the second phase, strain PAO1 produced quorum sensing (QS)-controlled secondary metabolites, among them the redox active pigment pyocyanin. Pyocyanin inhibited the enzyme aconitase of strain AH-1N through the production of reactive oxygen species causing a block of the citric acid cycle. This led to a massive acetate release by strain AH-1N, which supported substantial growth of strain PAO1. Strain AH-1N was finally inactivated by pyocyanin and presumably other secondary metabolites. Further investigation of this parasitic growth strategy of strain PAO1 revealed that, while catabolite repression of GlcNAc metabolism by acetate did not play a role, the action of isocitrate lyase was a key metabolic requirement for the transition into the second phase. Besides its role in acetate utilization, this enzyme was crucial for the utilization of GlcNAc. The ability to synthesize amino acids was a metabolic requirement of strain PAO1 as well. The overexpression of the QS effector protein PqsE regulating pyocyanin production could not restore formation of pyocyanin in auxotrophic mutants. P. aeruginosa possesses three QS systems. For the QS response of strain PAO1 in the co-culture, both the rhl and the 2-alkyl-4(1H)-quinolone system were crucial, whereas the las system was dispensable. Lack of the rhl signal synthase could be complemented by cross-talk with signals of strain AH-1N. By applying transposon mutagenesis and screening for mutants of strain PAO1 with defects in QS-regulated processes, we could identify several genes that were involved in the regulation of pyocyanin production. Among them were members of the gene cluster PA1415-PA1421, mutations of which led to a decreased production of pyocyanin and accelerated growth with the polyamine spermin.By employing our co-culture model systems to study interspecific interactions, we could identify strategies of bacteria that are likely to be important in their natural habitats. With regard to P. aeruginosa, our model system offers the possibility to study QS under conditions that are more ecologically relevant than in single culture

SpoT-Mediated Regulation and Amino Acid Prototrophy Are Essential for Pyocyanin Production During Parasitic Growth of Pseudomonas aeruginosa in a Co-culture Model System With Aeromonas hydrophila

The opportunistic pathogen Pseudomonas aeruginosa employs its complex quorum sensing (QS) network to regulate the expression of virulence factors such as pyocyanin. Besides cell density, QS in this bacterium is co-regulated by environmental cues. In this study, we employed a previously established co-culture model system to identify metabolic influences that are involved in the regulation of pyocyanin production in P. aeruginosa. In this co-culture consisting of P. aeruginosa and the chitinolytic bacterium Aeromonas hydrophila, parasitic growth of P. aeruginosa is strictly dependent on the production of pyocyanin. We could show that in this co-culture, pyocyanin production is likely induced by the stringent response mediated by SpoT in response to nutrient limitation. Pyocyanin production by stringent response mutants in the co-culture could not be complemented by overexpression of PqsE. Via transposon mutagenesis, several amino acid auxotrophic mutants were identified that were also unable to produce pyocyanin when PqsE was overexpressed or when complementing amino acids were present. The inability to produce pyocyanin even though PqsE was overexpressed was likely a general effect of amino acid auxotrophy. These results show the value of the co-culture approach to identify both extra- and intracellular metabolic influences on QS that might be important in infection processes as well

Parasitic growth of Pseudomonas aeruginosa in co-culture with the chitinolytic bacterium Aeromonas hydrophila

Polymer-degrading bacteria face exploitation by opportunistic bacteria that grow with the degradation products without investing energy into production of extracellular hydrolytic enzymes. This scenario was investigated with a co-culture of Aeromonas hydrophila and Pseudomonas aeruginosa with chitin as carbon, nitrogen and energy source. In single cultures, A. hydrophila could grow with chitin, while P. aeruginosa could not. Co-cultures with both strains had a biphasic course. In the first phase, P. aeruginosa grew along with A. hydrophila without affecting it. The second phase was initiated by a rapid inactivation of and a massive acetate release by A. hydrophila. Both processes coincided and were dependent on quorum sensing-regulated production of secondary metabolites by P. aeruginosa. Among these the redox-active phenazine compound pyocyanin caused the release of acetate by A. hydrophila by blocking the citric acid cycle through inhibition of aconitase. Thus, A. hydrophila was forced into an incomplete oxidation of chitin with acetate as end-product, which supported substantial growth of P. aeruginosa in the second phase of the co-culture. In conclusion, P. aeruginosa could profit from a substrate that was originally not bioavailable to it by influencing the metabolism and viability of A. hydrophila in a parasitic way

Table_2_SpoT-Mediated Regulation and Amino Acid Prototrophy Are Essential for Pyocyanin Production During Parasitic Growth of Pseudomonas aeruginosa in a Co-culture Model System With Aeromonas hydrophila.docx

<p>The opportunistic pathogen Pseudomonas aeruginosa employs its complex quorum sensing (QS) network to regulate the expression of virulence factors such as pyocyanin. Besides cell density, QS in this bacterium is co-regulated by environmental cues. In this study, we employed a previously established co-culture model system to identify metabolic influences that are involved in the regulation of pyocyanin production in P. aeruginosa. In this co-culture consisting of P. aeruginosa and the chitinolytic bacterium Aeromonas hydrophila, parasitic growth of P. aeruginosa is strictly dependent on the production of pyocyanin. We could show that in this co-culture, pyocyanin production is likely induced by the stringent response mediated by SpoT in response to nutrient limitation. Pyocyanin production by stringent response mutants in the co-culture could not be complemented by overexpression of PqsE. Via transposon mutagenesis, several amino acid auxotrophic mutants were identified that were also unable to produce pyocyanin when PqsE was overexpressed or when complementing amino acids were present. The inability to produce pyocyanin even though PqsE was overexpressed was likely a general effect of amino acid auxotrophy. These results show the value of the co-culture approach to identify both extra- and intracellular metabolic influences on QS that might be important in infection processes as well.</p

Interactions of bacteria with different mechanisms for chitin degradation result in the formation of a mixed-species biofilm

In this study, interactions between bacteria possessing either released or cell-associated enzymes for polymer degradation were investigated. For this, a co-culture of Aeromonas hydrophila strain AH-1N as an enzyme-releasing bacterium and of Flavobacterium sp. strain 4D9 as a bacterium with cell-associated enzymes was set up with chitin embedded into agarose beads to account for natural conditions, under which polymers are usually embedded in organic aggregates. In single cultures, strain AH-1N grew with embedded chitin, while strain 4D9 did not. In co-cultures, strain 4D9 grew and outcompeted strain AH-1N in the biofilm fraction. Experiments with cell-free culture supernatants containing the chitinolytic enzymes of strain AH-1N revealed that growth of strain 4D9 in the co-culture was based on intercepting N-acetylglucosamine from chitin degradation. For this, strain 4D9 had to actively integrate into the biofilm of strain AH-1N. This study shows that bacteria using different chitin degradation mechanisms can coexist by formation of a mixed-species biofilm

The guanidinobutyrase GbuA is essential for the alkylquinolone-regulated pyocyanin production during parasitic growth of Pseudomonas aeruginosa in co-culture with Aeromonas hydrophila

Jagmann N, Bleicher V, Busche T, Kalinowski J, Philipp B. The guanidinobutyrase GbuA is essential for the alkylquinolone-regulated pyocyanin production during parasitic growth of Pseudomonas aeruginosa in co-culture with Aeromonas hydrophila. ENVIRONMENTAL MICROBIOLOGY. 2016;18(10):3550-3564.The opportunistic pathogen Pseudomonas aeruginosa controls the production of virulence factors by quorum sensing (QS). Besides cell density, QS in P. aeruginosa is co-regulated by metabolic influences, especially nutrient limitation. Previously, a co-culture model system was established consisting of P. aeruginosa and the chitinolytic bacterium Aeromonas hydrophila, in which parasitic growth of P. aeruginosa is strictly dependent on the QS-controlled production of pyocyanin in response to nutrient limitation (Jagmann et al., 2010). In this study, the co-culture was employed to identify novel genes involved in the regulation of pyocyanin production. Via transposon mutagenesis, the gene gbuA encoding a guanidinobutyrase was identified, deletion of which led to a loss of pyocyanin production in co-cultures and to a reduced pyocyanin production in single cultures. Addition of the natural substrate of GbuA to the mutant strain enhanced the negative effect on pyocyanin production in single cultures. The gbuA mutant showed a reduced transcription of the pqsABCDE operon and could be complemented by PqsE overexpression and addition of alkylquinolone signal molecules. The strong effect of gbuA deletion on the QS-controlled pyocyanin production in co-cultures showed the value of this approach for the discovery of novel gene functions linking metabolism and QS in P. aeruginosa

Synthetic Escherichia coli-Corynebacterium glutamicum consortia for L-lysine production from starch and sucrose

Sgobba E, Stumpf AK, Vortmann M, et al. Synthetic Escherichia coli-Corynebacterium glutamicum consortia for L-lysine production from starch and sucrose. Bioresource Technology. 2018;260:302-310

Identification of a thiolase gene essential for β-oxidation of the acyl side chain of the steroid compound cholate in Pseudomonas sp. strain Chol1.

Bile salts such as cholate are steroid compounds occurring ubiquitously in the environment through excretion by animals. Cholate degradation by Pseudomonas sp. strain Chol1 is initiated by A-ring oxidation and β-oxidation of the acyl side chain. A transposon mutant of strain Chol1 was isolated that could not grow with cholate, but transformed it into several steroid compounds accumulating in culture supernatants. The main product was identified as (22E)-7α,12α-dihydroxy-3-oxochola-1,4,22-triene-24-oate (DHOCTO). A further compound was identified as 7α,12α,22-trihydroxy-3-oxochola-1,4-diene-24-oate (THOCDO). The structures of DHOCTO and THOCDO indicate that they are intermediates of the β-oxidation of the acyl side chain. The interrupted gene was named skt and had similarities to the 3-ketoacyl-CoA thiolase domain of the eukaryotic sterol carrier protein SCP-x. An skt mutant grew with intermediates of cholate degradation, from which the acyl side chain had been partly or completely removed. Growth with cholate was restored by an intact skt copy on a plasmid. These results strongly suggest that skt encodes a β-ketothiolase responsible for the cleavage of acetyl-CoA from the acyl side chain of cholate. Sequence comparisons revealed that other steroid-degrading bacteria such as Comamonas testosteroni contain genes encoding proteins very similar to Skt, suggesting a widespread role of this enzyme in bacterial steroid degradation