De novo sequencing of two novel peptides homologous to calcitonin-like peptides, from skin secretion of the Chinese Frog, Odorrana schmackeri

An MS/MS based analytical strategy was followed to solve the complete sequence of two new peptides from frog (Odorrana schmackeri) skin secretion. This involved reduction and alkylation with two different alkylating agents followed by high resolution tandem mass spectrometry. De novo sequencing was achieved by complementary CID and ETD fragmentations of full-length peptides and of selected tryptic fragments. Heavy and light isotope dimethyl labeling assisted with annotation of sequence ion series. The identified primary structures are GCD[I/L]STCATHN[I/L]VNE[I/L]NKFDKSKPSSGGVGPESP-NH2 and SCNLSTCATHNLVNELNKFDKSKPSSGGVGPESF-NH2, i.e. two carboxyamidated 34 residue peptides with an aminoterminal intramolecular ring structure formed by a disulfide bridge between Cys2 and Cys7. Edman degradation analysis of the second peptide positively confirmed the exact sequence, resolving I/L discriminations. Both peptide sequences are novel and share homology with calcitonin, calcitonin gene related peptide (CGRP) and adrenomedullin from other vertebrates. Detailed sequence analysis as well as the 34 residue length of both O. schmackeri peptides, suggest they do not fully qualify as either calcitonins (32 residues) or CGRPs (37 amino acids) and may justify their classification in a novel peptide family within the calcitonin gene related peptide superfamily. Smooth muscle contractility assays with synthetic replicas of the S–S linked peptides on rat tail artery, uterus, bladder and ileum did not reveal myotropic activity.

Doing Relationships and Sexuality Education with Young People in State Care

Background: Existing literature indicates that young people in state carehave particular sexual health needs that include addressing their social andemotional well-being, yet little has been published as to how thesecomponents of sex education are actually delivered by service-providers.Objective: To analyse the processes involved in delivering relationship andsexuality education to young people in state care from the perspectives ofa sample of service-providers with a role in sexual health care delivery.Design: Qualitative methodological strategy.Setting: Service-delivery sites at urban and rural locations in Ireland.Method: Twenty-two service-providers were interviewed in depth, and datawere analysed using a qualitative analytical strategy resembling modifiedanalytical induction.Findings: Participants proffered their perceptions and examples of theirpractices of sex education in relation to the following themes: (1)acknowledging the multi-dimensional nature of sexual health in the case ofyoung people in care; (2) personal and emotional development educationto address poor self-esteem, emotional disconnectedness and an inabilityto recognise and express emotions; (3) social skills’ education as part of arepertoire of competencies needed to negotiate relationships and safer sex;(4) the application of positive social skills embedded in everyday socialsituations; and (5) factual sexuality education.Conclusion: Insights into service providers’ perceptions of the multidimensionalnature of the sexual health needs of young people in statecare, and the ways in which these service-providers justified their practicemake visible the complex character of sex education and the degree of skillrequired to deliver it to those in state care.

Molecularly imprinted polymers for the extraction of imiquimod from biological samples using a template analogue strategy

Molecularly Imprinted Polymers (MIPs) against imiquimod, a highly potent immune response modifier used in the treatment of skin cancer, were synthesised using a template analogue strategy and were compared with imprints of the drug itself. An investigation of the complexation between the functional monomer and the template analogue revealed an association constant of 1,376 ± 122 M-1, significantly higher than previously reported values for similar systems. The binding characteristics of the synthesised imprinted polymers were evaluated and extremely strong binding for imiquimod was observed while imprinting factors as high as 17 were calculated. When applied as sorbents in solid-phase extraction of imiquimod from aqueous, urine and blood serum samples, clean extracts and recoveries up to 95% were achieved, and it is concluded that while imiquimod imprints exhibited higher capacity for the drug, template analogue imprints are more selective. The results obtained suggest potential applications of imiquimod imprints as sorbents in rapid extraction and monitoring of undesirable systemic release of the drug.