Engineering Dehydrated Amino Acid Residues in the Antimicrobial Peptide Nisin

The small antimicrobial peptide nisin, produced by Lactococcus lactis, contains the uncommon amino acid residues dehydroalanine and dehydrobutyrine and five thio ether bridges. Since these structures are posttranslationally formed from Ser, Thr, and Cys residues, it is feasible to study their role in nisin function and biosynthesis by protein engineering. Here we report the development of an expression system for mutated nisin Z (nisZ) genes, using nisin A producing L. lactis as a host. Replacement by site-directed mutagenesis of the Ser-5 codon in nisZ by a Thr codon, led to a mutant with a dehydrobutyrine instead of a dehydroalanine residue at position 5, as shown by NMR. Its antimicrobial activity was 2-10-fold lower relative to wild-type nisin Z, depending on the indicator strain used. In another mutagenesis study a double mutation was introduced in the nisZ gene by replacing the codons for Met-17 and Gly-18 by codons for Gln and Thr, respectively, as in the third lanthionine ring of the related antimicrobial peptide subtilin from Bacillus subtilis. This resulted in the simultaneous production of two mutant species, one containing a Thr residue and the other containing a dehydrobutyrine residue at position 18, both having different bacteriocidal properties.

Assessing demand for faecal sludge management (FSM) services in Freetown

‘Achieving sustained environmental health improvements in Freetown through faecal sludge management enterprises’ was a partnership project between Freetown City Council (FCC), International Water Association (IWA), and GOAL. This project aimed to improve faecal sludge management (FSM) through public – private partnerships and improved financial flows to ensure viability of businesses. A market assessment was conducted in Freetown which considered demand and supply for FSM services. This paper discusses findings from the household survey which was a key component of the market assessment. While the households felt that they were getting value for money for existing services, they were not satisfied with existing services and hence were willing to pay higher prices for improved services. This highlights the importance of quality service provision in relation to revenue generation. Improved pit emptying services was noted to be of high priority to households. In order to improve pit emptying services the FSM strategy proposed intermediate transfer stations and formation of a Sanitation Unit and Contact Centre within FCC

Sublingual allergen immunotherapy with a liquid birch pollen product in patients with seasonal allergic rhinoconjunctivitis with or without asthma

Background: Sublingual allergen immunotherapy (SLIT) has been demonstrated to be both clinically efficacious and safe. However, in line with the current regulatory guidance from the European Medicines Agency, allergen immunotherapy (AIT) products must demonstrate their efficacy and safety in pivotal phase III trials for registration. Objective: We sought to investigate the efficacy and safety of sublingual high-dose liquid birch pollen extract (40,000 allergy units native [AUN]/mL) in adults with birch pollen allergy. Methods: A randomized, double-blind, placebo-controlled, parallel-group multicenter trial was conducted in 406 adult patients with moderate-to-severe birch pollen-induced allergic rhinoconjunctivitis with or without mild-to-moderate controlled asthma. Treatment was started 3 to 6 months before the birch pollen season and continued during the season in 40 clinical study centers in 5 European countries. For primary end point assessment, the recommended combined symptom and medication score of the European Academy of Allergy and Clinical Immunology was used. Secondary end points included quality-of-life assessments, immunologic parameters, and safety. Results: Primary efficacy results demonstrated a significant (P < .0001) and clinically relevant (32%) reduction in the combined symptom and medication score compared with placebo after 3 to 6 months of SLIT. Significantly better rhinoconjunctivitis quality-of-life scores (P < .0001) and the patient's own overall assessment of his or her health status, including the visual analog scale score (Euro Quality of Life Visual Analogue Scale; P = .0025), were also demonstrated. In total, a good safety profile of SLIT was observed. Conclusion: This study confirmed both the clinical efficacy and safety of a sublingual liquid birch pollen extract in adults with birch pollen allergy in a pivotal phase III trial (EudraCT: 2013-005550-30; ClinicalTrials. gov: NCT02231307)

The adrenocortical response to synthetic ACTH following a trek to high altitude

Background: Gradual ascent to high altitude is typically associated with reduced resting aldosterone and unchanged cortisol, features that may facilitate acclimatization but are poorly understood. Aims: To investigate the cortisol and aldosterone response to adrenocorticotrophic hormone at altitude. Methods: Eleven subjects underwent a 250 µg short synacthen test at sea-level and again after trekking to 3600 m in Nepal. Cortisol and aldosterone were measured by conventional assay from blood samples taken immediately prior to the administration of synacthen (T0) and then 30 (T30) and 60 (T60) minutes later. Results: At 3600 m resting basal cortisol and aldosterone levels were both significantly lower than they were at sea-level (p=0.004, p=0.003 respectively). Cortisol values at T30 and T60 were no different between sea-level and 3600 m but the increment after synacthen was significantly (p=0.041) greater at 3600 m due to a lower basal value. Aldosterone at T30 and T60 was significantly lower (p=0.003 for both) at 3600 m than at sea-level and the increment following synacthen was also significantly (p=0.003) less at 3600 m. Conclusions: At 3600 m there appears to be a divergent adrenal response to synthetic adrenocorticotrophic hormone with an intact cortisol response but a reduced aldosterone response, relative to sea-level. This may reflect a specific effect of hypoxia on aldosterone synthesis and may be beneficial to acclimatization

Immunogenicity of standard and low dose vaccination using yeast-derived recombinant hepatitis B surface antigen in elderly volunteers

There is no conclusive evidence that age influences the response to vaccination against hepatitis B virus. We therefore studied the immunogenicity of yeast-derived rHBsAg vaccine in elderly volunteers. The study was conducted in the outpatient clinics of an academic and a regional hospital, in a rural family practice and in an urban community centre. We recruited 112 healthy volunteers aged 59 years and over, to whom 10 or 20 μg yeast-derived HBsAg was given at 0, 1 and 6 months. Anti-HBs titres were measured by radioimmunoassay at 2, 6 and 7 months. Responders and non-responders were compared using univariate non-parametric tests and multivariate logistic regression analysis. Of the 116 subjects who volunteered to take part in the study, 106 vaccinees completed it. The percentage of subjects with an anti-HBs titre ⩾10 1U1−1 at 7 months was 60% (95% confidence interval: 51–70%; geometric mean titre: 253 1U1−1). Of the factors studied, i.e. setting, age, sex, alcohol consumption, current medication and vaccine dose, the use of medication at the time of the first vaccination was the only independent factor related to the response to vaccination, with a response rate of 78% (95% confidence interval: 66–89%) in those without medication. In elderly subjects, the proportion with protective concentrations of anti-HBs after vaccination with 10 or 20 μg yeast-derived recombinant HBsAg in a standard scheme is lower than in healthy adolescents. Within the older age group studied here, the use of medication, probably reflecting general health, is the only significant factor influencing the response to vaccination

Hot, Sour and Breathless – Ocean under Stress.

ISBN 978-0-9519618-6-

Implementation of the participatory approach for supervisors to prevent sick leave: a process evaluation

To perform a process evaluation of a multifaceted strategy to implement the participatory approach for supervisors to prevent sick leave in three organisations. The implementation strategy incorporated a working group meeting with stakeholder representatives, supervisor training, and optional supervisor coaching. Context, recruitment, reach, dose delivered, dose received, fidelity, and satisfaction with the strategy were assessed at organisational and supervisor level using questionnaires and registration forms. At least 4 out of 6 stakeholders were represented in the working group meetings, and 11 % (n = 116) of supervisors could be reached. The working group meetings and supervisor training were delivered and received as planned and were well appreciated within all three organisations. Three supervisors made use of coaching. At 6-month follow-up, 11 out of 41 supervisors (27 %) indicated that they had applied the participatory approach at least one time. The implementation strategy was largely carried out as intended. However, reach of both supervisors and department managers should be improved. Future studies should consider targeting employees with the strategy

Collateral and Debt Maturity Choice. A Signaling Model

This paper derives optimal loan policies under asymmetric information where banks offer loan contracts of long and short duration, backed or unbacked with collateral. The main novelty of the paper is that it analyzes a setting in which high quality firms use collateral as a complementary device along with debt maturity to signal their superiority. The least-cost signaling equilibrium depends on the relative costs of the signaling devices, the difference in firm quality and the proportion of good firms in the market. Model simulations suggest a non-monotonic relationship between firm quality and debt maturity, in which high quality firms have both long-term secured debt and short-term secured or non-secured debt.

Magnetic resonance imaging-guided phase 1 trial of putaminal AADC gene therapy for Parkinson's disease.

ObjectiveTo understand the safety, putaminal coverage, and enzyme expression of adeno-associated viral vector serotype-2 encoding the complementary DNA for the enzyme, aromatic L-amino acid decarboxylase (VY-AADC01), delivered using novel intraoperative monitoring to optimize delivery.MethodsFifteen subjects (three cohorts of 5) with moderately advanced Parkinson's disease and medically refractory motor fluctuations received VY-AADC01 bilaterally coadministered with gadoteridol to the putamen using intraoperative magnetic resonance imaging (MRI) guidance to visualize the anatomic spread of the infusate and calculate coverage. Cohort 1 received 8.3 × 1011 vg/ml and ≤450 μl per putamen (total dose, ≤7.5 × 1011 vg); cohort 2 received the same concentration (8.3 × 1011 vg/ml) and ≤900 μl per putamen (total dose, ≤1.5 × 1012 vg); and cohort 3 received 2.6 × 1012 vg/ml and ≤900 μl per putamen (total dose, ≤4.7 × 1012 vg). (18)F-fluoro-L-dihydroxyphenylalanine positron emission tomography (PET) at baseline and 6 months postprocedure assessed enzyme activity; standard assessments measured clinical outcomes.ResultsMRI-guided administration of ascending VY-AADC01 doses resulted in putaminal coverage of 21% (cohort 1), 34% (cohort 2), and 42% (cohort 3). Cohorts 1, 2, and 3 showed corresponding increases in enzyme activity assessed by PET of 13%, 56%, and 79%, and reductions in antiparkinsonian medication of -15%, -33%, and -42%, respectively, at 6 months. At 12 months, there were dose-related improvements in clinical outcomes, including increases in patient-reported ON-time without troublesome dyskinesia (1.6, 3.3, and 1.5 hours, respectively) and quality of life.InterpretationNovel intraoperative monitoring of administration facilitated targeted delivery of VY-AADC01 in this phase 1 study, which was well tolerated. Increases in enzyme expression and clinical improvements were dose dependent. ClinicalTrials.gov Identifier: NCT01973543 Ann Neurol 2019;85:704-714