Perspectives on intranasal administration of corticosteroids:insights into different aspects related to patients, administration techniques and instructions

Intranasal administration of corticosteroids: patients, administration techniques and instructionsIntranasal corticosteroids (INCs) are used in allergic rhinitis and suppress allergic inflammation in the nasal mucosa. Adequate administration of INCs has received limited attention in clinical practice and scientific literature, whilst research about correct administration of asthma inhalers is widely available and has become a major focus area. Corine Rollema approaches different aspects related to treatment with INCs in her thesis. This thesis shows that there is sufficient research about the efficacy and safety of INCs. However, efficacy and safety may also depend on nasal particle deposition. According to Rollema's research it remains unclear whether, and to what extent, a set of administration variables affect deposition.In addition, patients apply certain habits for administration of INCs based on different instructional sources. The conducted research shows that various instructional sources including instructions in patient information leaflets, given by healthcare providers and in videos on YouTube® are incomplete, non-uniform or of insufficient quality.This thesis describes what characterizes current INC-users and what side effects may occur in clinical practice. In addition, the administration technique of patients was analyzed and knowledge about the administration of INCs was found to be insufficient. To successfully instruct children about the administration of INCs, this thesis indicates that an instructional video for children is an effective tool to improve administration.Rollema hopes that further research about the administration of INCs will lead to a combination of a well-substantiated administration technique and a patient-tailored instruction and evaluation, which may lead to optimal treatment with INCs

Uroflowmetry in males : Refrence values and clinical application in benign prostatic hypertrophy.

In this thesis the results are presented of a computerassisted study on uroflowmetry in males. Chapter I introduces the aim of the study and gives a short reviw. Urinary outflow obstruction due to begign prostatic hypertrophy (BPH) is a slowly progressive disease that initially gives rise to a few symptoms, but with prolonged negelct may eventually cause serious damage to the organs of the urinary tract. ... Summary and conclusion

Structure, Organization, and Expression of the lct Gene for Lacticin 481, a Novel Lantibiotic Produced by Lactococcus lactis

The structural gene for the lactococcal lantibiotic lacticin 481 (lct) has been identified and cloned using a degenerated 20-mer DNA oligonucleotide based on the amino-terminal 7 amino acid residues of the purified protein. The transcription of the lct gene was analyzed, and its promoter was mapped. DNA sequence analysis of the lct gene revealed an open reading frame encoding a peptide of 51 amino acids. Comparison of its deduced amino acid sequence with the amino-terminal sequence and the amino acid composition of lacticin 481 indicates that the 61-residue peptide is prelacticin 481, containing a 27-residue carboxyl-terminal propeptide and a 24-residue amino-terminal leader peptide which lacks the properties of a typical signal sequence and which is significantly different from the leaders of other lantibiotics. The predicted amino acid sequence of prolacticin 481 contains 3 cysteines, 2 serines, and 2 threonines which were not detectable in amino acid analyses of mature lacticin 481. Based on these results and on characterization by two-dimensional NMR techniques, a structural model is proposed in which 2 cysteine residues are involved in lanthionine and one in β-methyllanthionine formation, and a 4th threonine residue is dehydrated. This model predicts a molecular mass for lacticin 481 of 2,901, which is in excellent agreement with that obtained from mass spectrometry.

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.

Influence of Amino Acid Substitutions in the Nisin Leader Peptide on Biosynthesis and Secretion of Nisin by Lactococcus lactis

Structural genes for small lanthionine-containing antimicrobial peptides, known as lantibiotics, encode N-terminal leader sequences which are not present in the mature peptide, but are cleaved off at some stage in the maturation process. Leader sequences of the different lantibiotics share a number of identical amino acid residues, but they are clearly different from sec-dependent protein export signal sequences. We studied the role of the leader sequence of the lantibiotic nisin, which is produced and secreted by Lactococcus lactis, by creating site-directed mutations at various positions in the leader peptide sequence. Mutations at Arg-1 and Ala-4, but not at the conserved Pro-2, strongly affected the processing of the leader sequence and resulted in the extracellular accumulation of a biologically inactive precursor peptide. Amino acid analysis and 1H NMR studies indicated that the precursor peptide with an Ala-4 → Asp mutation contained a modified nisin structural part with the (mutated) unmodified leader sequence still attached to it. The Ala-4 → Asp precursor peptide could be activated in vitro by enzymatic cleavage with trypsin, liberating nisin. These results confirmed that cleavage of the leader peptide is the last step in nisin maturation and is necessary to generate a biologically active peptide. Several mutations, i.e. Pro-2 → Gly, Pro-2 → Val, Asp-7 → Ala, Lys-9 → Leu, Ser-10 → Ala/Ser-12 → Ala and Val-11 → Asp/Val-13 → Glu in the leader peptide did not have any detectable effect on nisin production and secretion, although some of them affected highly conserved residues. When mutations were created in the -18 to -15 region of the nisin leader peptide (i.e. Phe-18 → Leu, Leu-16 → Lys, Asp-15 → Ala), no secretion or intracellular accumulation could be detected of nisin or its precursors. This suggested that these conserved residues are involved in the maturation process and may interact with lantibiotic-specific modifying enzymes.

Observational study of administering intranasal steroid sprays by healthcare workers

OBJECTIVES: Patients with allergic rhinitis receive their information about administering intranasal corticosteroid sprays (INCS) from healthcare workers. Since the majority of patients does not administer these sprays correctly, we investigated whether healthcare workers know how to administer INCS. SETTINGS: We studied participants at their working place: pharmacy, outpatient clinic or general practitioner centre for emergencies. PARTICIPANTS: Pharmacist assistants, general practitioners, paediatricians and ear nose throat doctors. DESIGN: Observational study. All the participants demonstrated the administration technique with a spray device filled with water. PRIMARY OUTCOME: Number of steps of administration of INCS based on the established INCS protocol. SECONDARY OUTCOME: Number of five steps are labelled essential to obtain optimal distribution of the medication. RESULTS: Among the 75 participants, none performed all the steps correctly. The median of correctly performed steps in the protocol was 14 out of 29. A significantly better result was found among the pharmacist assistants. The essential steps were performed by 27 out of the 75 participants (36%). CONCLUSION: The majority of healthcare workers does not know how to administer INCS correctly. Patients could, therefore, receive incorrect and non-uniform instructions. The education of healthcare workers on how to administer INCS correctly may be an option for improvement