Coping with stress at clinical practice among nursing students

Študenti zdravstvene nege so tekom študija izpostavljeni številnim stresorjem, tako v izobraževalni ustanovi kot tudi na klinični praksi. Vodilo zaključnega dela je preučiti stresorje in kako se z njimi študenti zdravstvene nege soočajo ter spoprijemajo v času študija Zdravstvene nege. V zaključnem delu smo uporabili kvantitativno metodologijo raziskovanja. Raziskavo smo izvedli med študenti zdravstvene nege na eni izmed zdravstvenih fakultet za severovzhodu Slovenije, ki izvaja dodiplomski študijski program Zdravstvena nega. Kot instrument zbiranja podatkov smo uporabili vprašalnik sestavljen iz treh sklopov (demografski podatki, lestvici Perceived Stress Scale in Coping Behaviours Inventory). Rezultate raziskave smo statistično obdelali s pomočjo računalniških programov Microsoft Word in Excel 2019 ter IBM SPSS Statistics 26. V raziskavi je sodelovalo 98 študentov. Z raziskavo smo ugotovili, da študenti zdravstvene nege na klinični praksi najpogosteje doživljajo stres zaradi učiteljev in zdravstvenega osebja (2,75 ± 0,63) ter zaradi neskladja med teorijo in prakso (3,37 ± 0,92). Študenti zdravstvene nege se s stresom najpogosteje soočajo na način, da ostanejo pozitivni v dani situaciji (3,45 ± 0,61) ter da ohranijo pozitiven odnos pri soočanju z življenjskimi dogodki (3,91 ± 0,90). Pozitivizem predstavlja glavno vodilo pri soočanju s stresom. Študente zdravstvene nege je potrebno spodbujati k soočanju s stresom na pravilen način. Prav tako je potrebno klinične mentorje usposobiti, da nudijo ustrezno pomoč študentu, ko mu stres predstavlja težavo.Nursing students are exposed to a number of stressors during their study path, both in ther clinical practice and at educational institution. Stress cannot be avoided, but nursing students can deal with in the right way and reducing the negative stress that works on them. In the final thesis, we used quantitative research methodology. The research was conducted among nursing students from one of the faculties of health care in the area of northeastern Slovenia, which is pursuing an undergraduate nursing program. 98 nursing students participated in the survey. As a data collection instrument, we used a three-part questionnaire containing the Perceived Stress Scale (PSS) and Coping Behaviors Inventory (CBI). The survey results were statistically processed in Microsoft Word and Excel 2019 and IBM SPSS Statistics 26. The study found that nursing students in clinical practice most often experience stress due to teachers and medical staff (2.75 ± 0.63) and due to the mismatch between theory and practice (3.37 ± 0.92). Nursing students most often cope with stress by staying positive in a given situation (3.45 ± 0.61) and maintaining a positive attitude in coping with life events (3.91 ± 0.90). Nursing students are experiencing stressors in clinical practice, but they knew how to cope with them. Positivism is the their main guide in dealing with stress. Knowledge of stressors and coping with them presents opportunities for further research. Nurisng students need sto be encouraged to deal with stressors in the right way. Clinical mentors needs to be trained to offer appropriate assistance to students, when stress become a real problem for them

Stability and degradation pathways of N-nitroso-hydrochlorothiazide and the corresponding aryl diazonium ion

Despite the fact that it was put on the market more than 60 years ago, hydrochlorothiazide (HCT) is still one of the most important antihypertensive drugs. Due to its chemical structure, which contains the secondary aryl-alkyl-amino moiety, it is vulnerable to the formation of N-nitrosamine drug substance-related impurity (NDSRI) N-nitroso-hydrochlorothiazide (NO-HCT). In our study, we reveal that NO-HCT degrades rapidly at pH values 6 to 8. The main degradation products identified are formaldehyde, thiatriazine, and aminobenzenesulfonic acid derivative. Interestingly, degradation of NO-HCT at pH values from 5 to 1 is significantly slower and provides a different impurity profile when compared to the profile generated between pH 6 and 8. Specifically, between pH 1 and 5, HCT is observed as one of the key degradation products of NO-HCT in addition to formaldehyde and aminobenzenesulfonic acid. Moreover, at pH 1, the aminobenzenesulfonic acid derivative is transformed to the corresponding diazonium salt in approximately 3% yield with the nitrosyl cation, which is released during the decomposition of NO-HCT to HCT. This diazonium is highly unstable above pH 5. To verify that degradation of NO-HCT does not produce the corresponding diazonium salt that could be formed via metabolic activation of NO-HCT, this diazonium salt and its hydrolytic and reduction degradation products were synthesized and used as standards for the identification of species formed during the degradation of NO-HCT. This enabled us to confirm that the corresponding aryl diazonium salt, which would be obtained from metabolic activation of NO-HCT, is not observed in the NO-HCT degradation pathway. Our study also demonstrates that this diazonium salt is stable only in the presence of a large excess of strong mineral acid under anhydrous conditions. In the presence of water, it is instantaneously converted to an aminobenzenesulfonic acid derivative. These findings suggest that the NO-HCT should not be considered as a typical compound belonging to the cohort of the concern

Time course of whole-cell, DERA-catalyzed batch reactions.

<p>Reactions were performed using <i>E. coli</i> BL21 (DE3) pET30/<i>deoC</i> fermentation cultures directly (DERA specific activity = 232 kRFU s⁻¹ g⁻¹, WCW = 207 g L⁻¹). Results are given as mass concentrations obtained from GC-FID analysis. The measured quantity of a particular compound, with the exception of the stable 6-ring hemiacetals (<b>3</b>), represents the sum of the corresponding equilibrium forms (hydrate, aldehyde and acetal/hemiacetal), which exist under the reaction conditions. <b>A:</b> Reaction species data from reactions using 400 mmol L⁻¹ of <b>2g</b> and 840 mmol L⁻¹ of <b>1</b> are shown. <b>1</b> (▪, black), <b>3a</b> (▴, blue) <b>3g</b> (♦, green), <b>8g</b> (•, red), <b>10g</b> (Δ, orange) and <b>2g</b> (◊, brown). <b>B:</b> Reaction species data from reactions using 400 mmol L⁻¹ of <b>2b</b> and 840 mmol L⁻¹ of <b>1</b> are shown. <b>1</b> (▪, black), <b>3a</b> (▴, blue) <b>3b</b> (♦, green), <b>8b</b> (•, red), <b>10b</b> (Δ, orange), <b>2b</b> (◊, brown), 2,6-chloro-2,4-dideoxyhexose (□, grey). Concentration of the latter (Information S8) is evaluated based on the assumption, that the GC-FID response factor is similar to that of <b>3b</b>.</p

Time course of whole-cell, DERA-catalyzed, fed-batch reactions yielding 3g.

<p>Reaction species data from three independent experiments using (in total) 550 mmol L⁻¹ of <b>2g</b> and 1200 mmol L⁻¹ of <b>1</b> are shown. Whole-cell catalyst (<i>E. coli</i> BL21 (DE3) pET30/<i>deoC</i> high-density culture) with 217 kRFU s⁻¹ g⁻¹ DERA specific activity and 182 g L⁻¹ WCW was used. Results are given as molar concentrations obtained from GC-FID analysis. The measured quantity of a particular compound, with the exception of the stable 6-ring hemiacetals (<b>3</b>), represents the sum of the corresponding equilibrium forms (hydrate, aldehyde and acetal/hemiacetal) which exist under the reaction conditions. <b>1</b> (<b>▪</b>, black), <b>3a</b> (▴, blue) <b>3g</b> (♦, green), <b>8g</b> (•, red), <b>10g</b> (Δ, orange), <b>2g</b> (◊, brown), cumulative molarity of reaction species originating from <b>2g</b> (□, grey; sum of <b>2g</b>, <b>8g</b>, <b>10g</b> and <b>3g</b> ). Secondary vertical axis shows in %: residual DERA activity (□, violet), cumulative feed of <b>2g</b> (dotted line), cumulative feed of <b>1</b> (dashed line).</p

A Highly Productive, Whole-Cell DERA Chemoenzymatic Process for Production of Key Lactonized Side-Chain Intermediates in Statin Synthesis

<div><p>Employing DERA (2-deoxyribose-5-phosphate aldolase), we developed the first whole-cell biotransformation process for production of chiral lactol intermediates useful for synthesis of optically pure super-statins such as rosuvastatin and pitavastatin. Herein, we report the development of a fed-batch, high-density fermentation with <i>Escherichia coli</i> BL21 (DE3) overexpressing the native <i>E. coli deoC</i> gene. High activity of this biomass allows direct utilization of the fermentation broth as a whole-cell DERA biocatalyst. We further show a highly productive bioconversion processes with this biocatalyst for conversion of 2-substituted acetaldehydes to the corresponding lactols. The process is evaluated in detail for conversion of acetyloxy-acetaldehyde with the first insight into the dynamics of reaction intermediates, side products and enzyme activity, allowing optimization of the feeding strategy of the aldehyde substrates for improved productivities, yields and purities. The resulting process for production of ((2<i>S</i>,4<i>R</i>)-4,6-dihydroxytetrahydro-2<i>H</i>-pyran-2-yl)methyl acetate (acetyloxymethylene-lactol) has a volumetric productivity exceeding 40 g L⁻¹ h⁻¹ (up to 50 g L⁻¹ h⁻¹) with >80% yield and >80% chromatographic purity with titers reaching 100 g L⁻¹. Stereochemical selectivity of DERA allows excellent enantiomeric purities (<i>ee</i> >99.9%), which were demonstrated on downstream advanced intermediates. The presented process is highly cost effective and environmentally friendly. To our knowledge, this is the first asymmetric aldol condensation process achieved with whole-cell DERA catalysis and it simplifies and extends previously developed DERA-catalyzed approaches based on the isolated enzyme. Finally, applicability of the presented process is demonstrated by efficient preparation of a key lactol precursor, which fits directly into the lactone pathway to optically pure super-statins.</p></div

Time course of exemplary whole-cell, DERA-catalyzed, fed-batch reactions with ∼50 g L⁻¹ h⁻¹ volumetric productivity.

<p>Whole-cell catalyst (E. coli BL21 (DE3) pET30/deoC high-density culture) with 247 kRFU s⁻¹ g⁻¹ DERA specific activity and 215 g L⁻¹ WCW was used. Results are given as mass concentrations obtained from GC-FID analysis. The measured quantity of a particular compound, with the exception of the stable 6-ring hemiacetals (<b>3</b>), represents the sum of the corresponding equilibrium forms (hydrate, aldehyde and acetal/hemiacetal) which exist under the reaction conditions. <b>A:</b> Reaction species data from reaction using (in total) 700 mmol L⁻¹ of <b>2g</b> and 1540 mmol L⁻¹ of <b>1</b> are shown. <b>1</b> (▪, black), <b>3a</b> (▴, blue) <b>3g</b> (♦, green), <b>8g</b> (•, red), <b>10g</b> (Δ, orange), <b>2g</b> (◊, brown), acetic acid (□, grey). Secondary vertical axis shows in %: cumulative feed of <b>2g</b> (dotted line), cumulative feed of <b>1</b> (dashed line). <b>B:</b> Reaction species data from reaction using (in total) 700 mmol L⁻¹ of <b>2b</b> and 1540 mmol L⁻¹ of <b>1</b> are shown. <b>1</b> (▪, black), <b>3a</b> (▴, blue) <b>3b</b> (♦, green), <b>8b</b> (•, red), <b>10b</b> (Δ, orange), <b>2b</b> (◊, brown), acetic acid (□, grey), 2,6-chloro-2,4-dideoxyhexose (□, purple). Secondary vertical axis shows in %: cumulative feed of <b>2b</b> (dotted line), cumulative feed of <b>1</b> (dashed line).</p

DERA activity measurements of the whole-cell catalyst.

<p><b>A.</b> Fluorescence raw data for a DERA activity assay (dotted lines). Velocities for triplicate samples of the whole-cell catalyst were measured for 7 different loads (<i>b–h,</i> 3.16 µg–26.9 µg in 3.96 µg increments) of biomass. After normalization with the blank (<i>a</i>), maximum slopes were determined for each sample and averaged (solid lines) to yield velocity for a given biomass load. <b>B.</b> Velocity vs. biomass load plot. The first 5 points are taken for the specific activity calculation. Linear regression: y = 0.2366x+0.2073 R² = 0.9936 <b>C.</b> Comparison of velocities measured for cell-free lysate spiked with increasing loads of biomass. <b>D.</b> Validation of linearity of the activity assay within samples with constant biomass. The whole-cell catalyst <i>E. coli</i> BL21 (DE3) pET30/<i>deoC</i> was mixed with w.t. <i>E. coli</i> BL21 (DE3) biomass (•). Linear regression: y = 248.94x+1.3840, R² = 0.9995. In parallel, sonicated and cleared samples were measured (□). Linear regression: y = 235.00x+2.6433, R² = 0.9989.</p

Current art in the direct synthesis of optically pure super-statins from the lactol precursors.

<p>Current art in the direct synthesis of optically pure super-statins from the lactol precursors.</p

Inactivation of DERA whole-cell catalyst and DERA cell-free lysate with various aldehydes.

<p>Samples were treated with 75 mM, 150 mM and 225 mM substrate aldehydes for 15 minutes prior to the activity assay. The specific DERA activity was 226.8 kRFU s⁻¹ g⁻¹ and 226.6 kRFU s⁻¹ g⁻¹ for the whole-cell catalyst and for the cell-free lysate, respectively. Residual activities are given relative to non-treated whole-cell catalyst. Aldehydes used were acetaldehyde (A), <b>2b</b> (B) and <b>2g</b> (C).</p

The optimization of lactol (<b>3g</b>) oxidation yielding lactone <b>15</b>.

<p>The optimization of lactol (<b>3g</b>) oxidation yielding lactone <b>15</b>.</p