Spending time in the forest or the field: qualitative semi-structured interviews in a randomized controlled cross-over trial with highly sensitive persons.

Introduction: The effects of spending time in forests have been subject to investigations in various countries around the world. Qualitative comparisons have been rarely done so far. Methods: Sixteen healthy highly sensitive persons (SV12 score ≥ 18) aged between 18 and 70 years were randomly assigned to groups spending 1 h in the forest and in the field at intervals of one week. Semi-structured interviews were conducted after each intervention and analyzed using a mixed-methods approach of content analysis and grounded theory. Results: Both natural environments induced feelings of inner calmness, inner cleansing, joy, freedom, connectedness, strengthening qualities, and heightened body awareness. The forest environment additionally offered emotional shelter, and showed advantages in promoting inner strength and self-concentration. Discussion: People with previous negative experiences in the forest may feel safer in fields because of the wider view and better overview. Important preconditions are enough time and the absence of a judgmental authority. The two environments induced in part different but also similar emotions which might be useful to promote psychological well-being differentially.

Mindfulness and compassion-oriented practices at work reduce distress and enhance self-care of palliative care teams: a mixed-method evaluation of an "on the job" program.

BACKGROUND: Maintaining a sense of self-care while providing patient centered care, can be difficult for practitioners in palliative medicine. We aimed to pilot an "on the job" mindfulness and compassion-oriented meditation training for interdisciplinary teams designed to reduce distress, foster resilience and strengthen a prosocial motivation in the clinical encounter. METHODS: Our objective was to explore the feasibility and effectiveness of this newly developed training. The study design was an observational, mixed-method pilot evaluation, with qualitative data, self-report data, as well as objective data (cortisol) measured before and after the program. Twenty-eight staff members of an interdisciplinary palliative care team participated in the 10-week training conducted at their workplace. Measures were the Perceived Stress Questionnaire, the Maslach Burnout Inventory, the somatic complaints subscale of the SCL-90-R, the Emotion Regulation Skills Questionnaire, the Hospital Anxiety and Depression Scale, and a Goal Attainment Scale that assessed two individual goals. Semi-structured interviews were employed to gain insight into the perceived outcomes and potential mechanisms of action of the training. T-tests for dependent samples were employed to test for differences between baseline and post-intervention. RESULTS: Significant improvements were found in two of three burnout components (emotional exhaustion and personal accomplishment), anxiety, stress, two emotional regulation competences and joy at work. Furthermore, 85% of the individual goals were attained. Compliance and acceptance rates were high and qualitative data revealed a perceived enhancement of self-care, the integration of mindful pauses in work routines, a reduction in rumination and distress generated in the patient contact as well as an enhancement of interpersonal connection skills. An improvement of team communication could also be identified. CONCLUSIONS: Our findings suggest that the training may be a feasible, effective and practical way of reducing caregiver-distress and enhancing the resources of palliative care teams.

Expression of recombinant staphylokinase in the methylotrophic yeast Hansenula polymorpha.

BACKGROUND: Currently, the two most commonly used fibrinolytic agents in thrombolytic therapy are recombinant tissue plasminogen activator (rt-PA) and streptokinase (SK). Whereas SK has the advantage of substantially lower costs when compared to other agents, it is less effective than either rt-PA or related variants, has significant allergenic potential, lacks fibrin selectivity and causes transient hypotensive effects in high dosing schedules. Therefore, development of an alternative fibrinolytic agent having superior efficacy to SK, approaching that of rt-PA, together with a similar or enhanced safety profile and advantageous cost-benefit ratio, would be of substantial importance. Pre-clinical data suggest that the novel fibrinolytic recombinant staphylokinase (rSAK), or related rSAK variants, could be candidates for such development. However, since an efficient expression system for rSAK is still lacking, it has not yet been fully developed or evaluated for clinical purposes. This study's goal was development of an efficient fermentation process for the production of a modified, non-glycosylated, biologically active rSAK, namely rSAK-2, using the well-established single cell yeast Hansenula polymorpha expression system. RESULTS: The development of an efficient large scale (80 L) Hansenula polymorpha fermentation process of short duration for rSAK-2 production is described. It evolved from an initial 1mL HTP methodology by successive scale-up over almost 5 orders of magnitude and improvement steps, including the optimization of critical process parameters (e.g. temperature, pH, feeding strategy, medium composition, etc.). Potential glycosylation of rSAK-2 was successfully suppressed through amino acid substitution within its only N-acetyl glycosylation motif. Expression at high yields (≥ 1g rSAK-2/L cell culture broth) of biologically active rSAK-2 of expected molecular weight was achieved. CONCLUSION: The optimized production process described for rSAK-2 in Hansenula polymorpha provides an excellent, economically superior, manufacturing platform for a promising therapeutic fibrinolytic agent.

Subjective concepts of chronically ill patients using distant healing.

Distant healing procedures consist of benevolent intentions, often taking the form of prayers for a patient. Despite inconclusive evidence regarding distant healing, prayers are a widespread health-related technique. We studied subjective concepts of distant healing in 17 patients suffering from chronic fatigue syndrome and multiple chemical sensitivity who were given distant healing during a randomized controlled trial. We applied reconstructive interview analysis when analyzing the results. The overall theme was the tension between mainstream medicine and the immaterial healing procedure. Several components highlighted this tension: (a) patterns of legitimizing the use of distant healing, (b) distant healing and the social setting, (c) integrating distant healing into their belief system, and (d) reconstruction of effects by means of hindsight. The interviews showed that patients felt the need to legitimize having tried distant healing. They had to bear the full ambiguity of biomedicine being in competition with distant healing, though also experiencing distant healing as giving support.

Characteristics of zinc transport by two bacterial cation diffusion facilitators from Ralstonia metallidurans CH34 and Escherichia coli.

CzcD from Ralstonia metallidurans and ZitB from Escherichia coli are prototypes of bacterial members of the cation diffusion facilitator (CDF) protein family. Expression of the czcD gene in an E. coli mutant strain devoid of zitB and the gene for the zinc-transporting P-type ATPase zntA rendered this strain more zinc resistant and caused decreased accumulation of zinc. CzcD, purified as an amino-terminal streptavidin-tagged protein, bound Zn2+, Co2+, Cu2+, and Ni2+ but not Mg2+, Mn2+, or Cd2+, as shown by metal affinity chromatography. Histidine residues were involved in the binding of 2 to 3 mol of Zn2+ per mol of CzcD. ZitB transported 65Zn2+ in the presence of NADH into everted membrane vesicles with an apparent Km of 1.4 microM and a Vmax of 0.57 nmol of Zn2+ min(-1) mg of protein(-1). Conserved amino acyl residues that might be involved in binding and transport of zinc were mutated in CzcD and/or ZitB, and the influence on Zn2+ resistance was studied. Charged or polar amino acyl residues that were located within or adjacent to membrane-spanning regions of the proteins were essential for the full function of the proteins. Probably, these amino acyl residues constituted a pathway required for export of the heavy metal cations or for import of counter-flowing protons.

Identification of a regulatory pathway that controls the heavy-metal resistance system Czc via promoter czcNp in Ralstonia metallidurans.

The CzcCBA cation-proton-antiporter is the most complicated and efficient heavy-metal resistance system known today and is essential for survival of Ralstonia metallidurans at high cobalt, zinc, or cadmium concentrations. Expression of Czc is tightly controlled by the complex interaction of several regulators. Double- and multiple-deletion studies demonstrated that four regulators encoded downstream of the czcCBA operon, CzcD, CzcS, CzcR and the newly identified CzcE, are involved in, but not essential for metal-dependent induction of czc. These proteins control expression of the czcNICBA region from the promoter czcNp. Northern analysis showed that czcDRS was transcribed as czcDR-specific and czcDRS-specific mRNAs. Transcription of czcE occurred independently of czcDRS transcription and was induced by zinc. CzcE is a periplasmic protein as indicated by phoA fusions. CzcE was purified and identified as a metal-binding protein. These data demonstrate that the transport protein CzcD, the two-component regulatory system CzcR, CzcS, and the periplasmic metal-binding protein CzcE exert metal-dependent control of czcNICBA expression via regulation of czcNp activity.

First step towards a quantitative model describing Czc-mediated heavy metal resistance in Ralstonia metallidurans.

Quantitative models were derived to explain heavy metal resistance in Ralstonia metallidurans. A deltaczcA deletion of the gene for the central component of the Co2+/Zn2+/Cd2+ efflux system CzcCBA combined with the expression level of czcCBA as studied with a phi(czcC-lacZ-czcBA) operon fusion demonstrated that CzcCBA was the only prerequisite for resistance to Co2+/Zn2+/Cd2+ at concentrations of 200 microM and above. The cellular content of the CzcA protein (resistance nodulation cell division protein family RND) determined by Western blot was used to model the CzcCBA expression level as the response to various metal concentrations. These data and experimentally determined uptake velocities were used to derive a flow equilibrium model that describes the cytoplasmic content c(i) of the cells as an interaction between cation uptake and CzcCBA-mediated efflux. Alternatively, binding of heavy metals to inactivated R. metallidurans cells was described with a modified Freundlich's equation. The metal content of growing R. metallidurans cells was determined and compared to the predictions of both models. High amounts of zinc precipitates. exclusively formed by living cells, prevented a model validation for zinc. An additional net efflux activity let to lower amounts of cell-bound Co2+ than predicted. The flow equilibrium model described cadmium resistance sufficiently for R. metallidurans growing in the presence of 0.2-1 mM Cd2+. Description of cadmium resistance in early stationary cells requires the binding model in addition to the flow equilibrium model. Thus, it was possible to simulate physiological events in growing cells by quantitative models that are derived from the biochemical data of the interacting transport proteins.

Interplay of the Czc system and two P-type ATPases in conferring metal resistance to Ralstonia metallidurans.

Cadmium and zinc are removed from cells of Ralstonia metallidurans by the CzcCBA efflux pump and by two soft-metal-transporting P-type ATPases, CadA and ZntA. The czcCBA genes are located on plasmid pMOL30, and the cadA and zntA genes are on the bacterial chromosome. Expression of zntA from R. metallidurans in Escherichia coli predominantly mediated resistance to zinc, and expression of cadA predominantly mediated resistance to cadmium. Both transporters decreased the cellular content of zinc or cadmium in this host. In the plasmid-free R. metallidurans strain AE104, single gene deletions of cadA or zntA had only a moderate effect on cadmium and zinc resistance, but zinc resistance decreased 6-fold and cadmium resistance decreased 350-fold in double deletion strains. Neither single nor double gene deletions affected zinc resistance in the presence of czcCBA. In contrast, cadmium resistance of the cadA zntA double mutant could be elevated only partially by the presence of CzcCBA. lacZ reporter gene fusions indicated that expression of cadA was induced by cadmium but not by zinc in R. metallidurans strain AE104. In the absence of the zntA gene, expression of cadA occurred at lower cadmium concentrations and zinc now served as an inducer. In contrast, expression of zntA was induced by both zinc and cadmium, and the induction pattern did not change in the presence or absence of CadA. However, expression of both genes, zntA and cadA, was diminished in the presence of CzcCBA. This indicated that CzcCBA efficiently decreased cytoplasmic cadmium and zinc concentrations. It is discussed whether these data favor a model in which the cations are removed either from the cytoplasm or the periplasm by CzcCBA.

Functional analysis of the Escherichia coli zinc transporter ZitB.

The membrane transporter ZitB responsible for Zn(II) efflux in Escherichia coli was studied by site-directed mutagenesis to elucidate the function of individual amino acid residues. Substitutions of several charged or polar residues, H53, H159, D163 and D186, located in predicted transmembrane domains resulted in loss of ZitB function. In contrast, neither the amino-terminal nor the carboxy-terminal regions, both histidine-rich, were required for function.