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Abstract: In this paper, microfluidic based system for amperometric detection of CRP is described. The amperometric sensors were designed to operate in a microfluidic system for immunoenzymatic test (ELISA). The developed microfluidic system with integrated Pt amperometric sensors is suitable for determination of CRP in the lowest concentration range valuable for clinical diagnosis - 0.1mg/L to 1mg/L. [Copyright &y& Elsevier]

In this paper, we discuss dendrimer usage in enzyme-based electrochemical biosensors, particularly with respect to biomolecule loading on the sensing surface. A novel approach to design bioactive layers with immobilized enzymes for electrochemical biosensors using the surface plasmon resonance (SPR) method in combination with electrochemical impedance spectroscopy was presented. The gold surface was modified with linear linkers (various mercaptoalkanoic acids and aminoalkanethiols) and poly(amidoamine) dendrimers from the first- to fifth-generation. The best functionalization procedure was established by detailed SPR studies and transferred onto gold electrodes to electrochemically examine the model enzymatic reaction catalysed by glutamate oxidase. In the case of the chronoamperometric method, the determined sensitivity was 3.36 ± 0.08 μA∙mM -1 , and the low limit of detection (LOD) was 1.52 μM. Comparing the sensitivity and LOD obtained for CV measurements, the values of these parameters were 2.5 times higher and 4 times lower, respectively, for the fourth-generation dendrimer-based biosensor and the biosensor with a linear linker. The positive impact of the dendrimer interlayer on the long-term enzyme activity was also confirmed. The research results indicate the possibility of a significant increase in the sensor response using the dendrimer itself without enriching it with electrochemical components., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Marcin Urbanowicz reports financial support was provided by National Science Centre Poland., (Copyright © 2023 Elsevier B.V. All rights reserved.)

To get a better insight into the antimicrobial potency of short cationic lipopeptides, 35 new entities were synthesized using solid phase peptide strategy. All newly obtained lipopeptides were designed to be positively charged from +1 to +4. This was achieved by introducing basic amino acid - lysine - into the lipopeptide structure and had a hydrophobic fatty acid chain attached. Lipopeptides were subjected to microbiological tests using reference strains of Gram-negative bacteria: Escherichia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa , Gram-positive bacteria: Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Enterococcus faecalis , and fungi: Candida albicans, Candida tropicalis, Aspergillus brasiliensis . The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimal fungicidal concentration (MFC) were established for each strain. The toxicity toward human cells was determined by hemolysis tests via minimum hemolytic concentration (MHC) determination. The effect of the trifluoroacetic acid (TFA) counter ion on the antimicrobial activity of lipopeptides was also examined by its removing and performing the antimicrobial tests using counter ion-free compounds. The study shows that lipopeptides are more potent against Gram-positive than Gram-negative strains. It was revealed that positive charge equals at least +2 is a necessary condition to observe significant antimicrobial activity, but only when it is balanced with a proper length of hydrophobic fatty acid chain. The hemolytic activity of lipopeptides strongly depends on amino acid composition of the hydrophilic portion of the molecule as well as fatty acid chain length. Compounds endowed with a greater positive charge were more toxic to human erythrocytes. This should be considered during new lipopeptide molecules design. Our studies also revealed the TFA counter ion has no significant effect on the antimicrobial behavior of cationic lipopeptides.

Today microbial drug resistance has become a serious problem not only within inpatient setting but also within outpatient setting. Repeated intake and unnecessary usage of antibiotics as well as the transfer of resistance genes are the most important factors that make the microorganisms resistant to conventional antibiotics. A large number of antimicrobials successfully used for prophylaxis and therapeutic purposes have now become ineffective [1, 2]. Therefore, new molecules are being studied to be used in the treatment of various diseases. Some of these molecules are structural compounds based on a combination of peptides, for example, naturally occurring endogenous peptide antibiotics and their synthetic analogues or molecules designed de novo using QSAR (quantitative structureproperty relationships)-based methods [3]. Trying to exploit numerous advantages of antimicrobial peptides such as high potency and selectivity, broad range of targets, potentially low toxicity and low accumulation in tissues, pharmaceutical industry aims to develop them as commercially available drugs and appropriate clinical trials are being conducted [4]. In this paper we define clinical trials steps and describe current status of several antimicrobial peptides under clinical development as well as briefly depict peptide drug formulation.

The widespread use of contact lenses is associated with several complications, including ocular biofilm-related infections. They are very difficult to manage with standard antimicrobial therapies, because bacterial growth in a biofilm is associated with an increased antibiotic resistance. The principal aim of this study was to evaluate the efficacy of antimicrobial peptides (AMPs) in eradication of bacterial biofilms formed on commercially available contact lenses. AMPs were synthesized according to Fmoc/tBu chemistry using the solid-phase method. Minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of the compounds were determined. Anti-biofilm activity of the antimicrobial peptides determined at different temperatures (25 °C and 37 °C) were compared with the effectiveness of commercially available contact lens solutions. All of the tested compounds exhibited stronger anti-biofilm properties as compared to those of the tested lens solutions. The strongest activity of AMPs was noticed against Gram-positive strains at a temperature of 25 °C., Conclusions: The results of our experiments encourage us toward further studies on AMPs and their potential application in the prophylaxis of contact lens-related eye infections.

Staphylococcus aureus (SA) colonizes the vast majority of patients with atopic dermatitis (AD). Its resistance to antibiotics and ability to form biofilms are the main origins of therapeutic complications. Endogenous antimicrobial peptides (AMPs) exhibit strong activity against SA, including antibiotic resistant strains as well as bacteria existing in biofilm form. The purpose of the present work was to determine the antistaphylococcal activity of two amphibian peptides against SA isolated from patients with AD. The AMPs demonstrated permanent activity towards strains exposed to sublethal concentrations of the compounds and significantly stronger antibiofilm activity in comparison to that of conventional antimicrobials. The results suggest the potential application of amphibian AMPs as promising antistaphylococcal agents for the management of skin infections.

Peptide-based drugs are promising group of compounds which are characterized by specificity to their in vivo targets and high potency of action (antineoplastic, immunoregulatory, antibacterial). The peptides, however, involve a relatively high risk of allergic reactions that are not predictable on the basis of their sequence and chemical properties. In this study, peripheral blood was obtained from 53 patients including 38 hypersensitive patients and 15 control patients. Basophil activation stimulated by two antibacterial peptides (camel, citropin 1.1), and acetylsalicylic acid was assessed by means of BAT (basophil activation test). Basophil activation stimulated by camel occurred in 7 of 38 patients with hypersensitivity (18.42%) as well as in 2 of 15 control patients (13.33%). Basophils were activated by citropin 1.1 in 7 of 38 hypersensitive patients (18.42%) and in none of the control patients. Using the Structural Database of Allergenic Proteins, we confirmed that the examined peptides share some structural similarities with common environmental allergens. Therefore, the cross-reactivity between potentially present anti-allergen IgE with examined peptides cannot be excluded. Our study proved that BAT, together with other biological tests and specific databases of allergenic compounds, may serve as an initial selection of new active peptides and proteins., (© 2015 John Wiley & Sons A/S.)

Diosgenyl 2-amino-2-deoxy-β-D-glucopyranoside is a synthetic saponin exhibiting attractive pharmacological properties. Different pathways tested by us to obtain this glycoside are summarized here. Moreover, the synthesis of N-alkyl and N,N-dialkyl derivatives of the glucopyranoside is presented. Evaluation of antibacterial and antifungal activities of these derivatives indicates that they have no inhibitory activity against Gram-negative bacteria, whereas many of the tested N-alkyl saponins were found to inhibit the growth of Gram-positive bacteria and human pathogenic fungi.

Lactoferrin (LF) is a naturally occurring antimicrobial peptide that is cleaved by pepsin to lactoferricin (LFcin). LFcin has an enhanced antimicrobial activity as compared to that of LF. Recently several hetero- and homodimeric antimicrobial peptides stabilized by a single disulfide bond linking linear polypeptide chains have been discovered. We have demonstrated that the S-S bond heterodimerization of lipopeptide Laur-Orn-Orn-Cys-NH 2 (peptide III) and the synthetic N -terminal peptide of human lactoferricin (peptide I) yields a dimer (peptide V), which is almost as microbiologically active as the more active monomer and at the same time it is much less toxic. Furthermore, it has been found that the S-S bond homodimerization of both peptide I and peptide III did not affect antimicrobial and haemolytic activity of the compounds. The homo- and heterodimerization of peptides I and III resulted in either reduction or loss of antifungal activity. This work suggests that heterodimerization of antimicrobial lipopeptides via intermolecular disulfide bond might be a powerful modification deserving consideration in the design of antimicrobial peptides.

Novel organic compounds found in the cuticular and internal lipids of medically important flies were identified. Uracil, 9-tricosene, 1-oleoyl glycerol, dimethyl suberate and butyl stearate were tested for their potential antifungal activity. Minimal inhibitory concentrations of the compounds against reference strains of fungi were determined. Uracil and dimethyl suberate slightly inhibited the growth of entomopathogenic fungi. The cuticular and internal lipids of Calliphora vicina, Calliphora vomitoria, Sarcophaga carnaria and Musca domestica were studied by gas chromatography (GC) combined with mass spectrometry (GC/MS). A comparison of the lipid extracts between the preimaginal and mature stages showed adults flies contained a higher total content of the identified components. Furthermore, their amounts distinctly predominated in the internal lipids of all the species. The amount of 9-tricosene was the highest in adults of C. vicina, while the larvae and pupae had a definitively lower amount of this compound. Uracil was found to be the most abundant component in extracts obtained from C. vomitoria especially in the internal lipids of adults. 1-oleoyl glycerol was detected in all of the examined species of flies. It was most abundant in the internal extracts isolated from the larvae of C. vicina and the pupae of C. vomitoria. Suberic acid dimethyl ester was found in the larval and pupal internal lipids of C. vicina and S. carnaria in low amounts. Butyl stearate was identified only in the internal lipids of the larvae and adults of houseflies., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

In this work, the self-organization and the behavior of the surfactant-like peptides in the presence of biological membrane models were studied. The studies were focused on synthetic palmitic acid-containing lipopeptides, C16-KK-NH2 (I), C16-KGK-NH2 (II) and C16-KKKK-NH2 (III). The self-assembly was explored by molecular dynamics simulations using a coarse-grained force field. The critical micellar concentration was estimated by the surface tension measurements. The thermodynamics of the peptides binding to the anionic and zwitterionic lipids were established using isothermal titration calorimetry (ITC). The influence of the peptides on the lipid acyl chain ordering was determined using FTIR spectroscopy. The compounds studied show surface-active properties with a distinct CMC over the millimolar range. An increase in the steric and electrostatic repulsion between polar head groups shifts the CMC toward higher values and reduces the aggregation number. An analysis of the peptide-membrane binding revealed a unique interplay between the initial electrostatic and the subsequent hydrophobic interactions enabling the lipopeptides to interact with the lipid bilayer. In the case of C16-KKKK-NH2 (III), compensation of the electrostatic and hydrophobic interactions upon binding to the anionic membrane has been suggested and consequently no overall binding effects were noticed in ITC thermograms and FTIR spectra., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Cationic amino acid-based surfactants were synthesized via solid phase peptide synthesis and terminal acylation of their α and ε positions with saturated fatty acids. Five new lipopeptides, N-α-acyl-N-ε-acyl lysine analogues, were obtained. Minimum inhibitory concentration and minimum bactericidal (fungicidal) concentration were determined on reference strains of bacteria and fungi to evaluate the antimicrobial activity of the lipopeptides. Toxicity to eukaryotic cells was examined via determination of the haemolytic activities. The surface-active properties of these compounds were evaluated by measuring the surface tension and formation of micelles as a function of concentration in aqueous solution. The cationic surfactants demonstrated diverse antibacterial activities dependent on the length of the fatty acid chain. Gram-negative bacteria and fungi showed a higher resistance than Gram-positive bacterial strains. It was found that the haemolytic activities were also chain length-dependent values. The surface-active properties showed a linear correlation between the alkyl chain length and the critical micelle concentration.

Fatty acids as components of cuticular lipids of insects play a significant role in antifungal in protection against fungal infection. The chemical composition of cuticular and internal extracts obtained from all developmental stages of flesh flies Sarcophaga carnaria was identified. The fatty acids were detected using gas chromatography coupled with mass spectrometry and the most abundant for all examined stages were: 18:1 > 16:0 > 16:1 > 18:0 > 18:2. Polyunsaturated fatty acids (PUFA) C20 were found in both, cuticular and internal extracts. GC-MS analysis showed higher relative content of PUFA in adults than in preimaginal stages. Fatty acids alone as well as their cuticular and internal extracts obtained from larvae, pupae male and female of S. carnaria were tested according to their potential antimicrobial activity against entomopathogenic fungi: Paecilomyces lilacinus, Paecilomyces fumosoroseus, Lecanicillium lecanii, Metarhizium anisopliae, Beauveria bassiana (Tve-N39) and B. bassiana (Dv-1/07). FA presented diverse antimicrobial activity depending on the length of the chain and the presence of unsaturated bonds. Short chain and unsaturated FA (6:0, 11:0, 13:0) have shown significantly stronger activity against fungi but they were detected in lower concentrations. PUFA inhibit fungal growth more effectively than unsaturated long chain fatty acids. Cuticular and internal extracts of all living forms of S. carnaria exhibited approximately equal activity against tested entomopathogenic fungi. We presumed that the most abundant saturated long chain FA and additionally PUFA founded in our analysis are involved in protecting the flies against fungal infection., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Recently, the topic of biofilm has met a huge interest of researchers owing to a significant role played by this microbial life form in severe infections. These well organised three-dimensional microbial communities are characterized by a strong resistance to antimicrobials. Biofilms significantly contribute to morbidity and mortality as related infections are very difficult to treat due to their tendency to relapse after the withdrawal of antibiotics. According to the literature, antimicrobial peptides (AMPs) have a high potential as future antibiofilm agents. AMPs can influence various stages of biofilm formation and exhibit antimicrobial activity against a broad spectrum of microorganisms including multi-drug resistant strains. The purpose of the present study was to determine the activity of antimicrobial peptides against biofilms formed by a variety of bacterial strains. To do this, the following antimicrobial peptides were synthesized: Citropin 1.1, Lipopeptides Palm-KK-NH2 and Palm-RR-NH2, Omiganan, Pexiganan and Temporin A. Antimicrobial activity of the compounds and conventional antibiotics was determined for planktonic cells and biofilms formed by reference strains of Gram-positive (Staphylococcus aureus, S. epidermidis, Streptococcus pneumoniae, Streptococcus pyogenes) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis) bacteria. AMPs exhibited a strong antibacterial activity against Gram-positive strains, while Gram-negative bacteria were less susceptible. Antimicrobial activity of the tested peptides against biofilms formed by Gram-positive organisms was significantly stronger as compared to that of conventional antimicrobials.

Over the last decade, the growing number of multidrug resistant strains limits the use of many of the currently available chemotherapeutic agents. Furthermore, bacterial biofilm, due to its complex structure, constitutes an effective barrier to conventional antibiotics. The in vitro activities of naturally occurring peptide (Citropin 1.1), chemically engineered analogue (Pexiganan), newly-designed, short amino-acid derivatives (Pal-KK-NH2, Pal-KKK-NH2, Pal-RRR-NH2) and six clinically used antimicrobial agents (Gatifloxacin, Ampicilin, Cefotaxime, Ceftriaxone, Cefuroxime and Cefalexin) were investigated against planktonic cells and mature biofilm of multidrug-resistant Providencia stuartii strains, isolated from urological catheters. The MICs, MBCs values were determined by broth microdilution technique. Inhibition of biofilm formation by antimicrobial agents as well as biofilm susceptibility assay were tested using a surrogate model based on the Crystal Violet method. The antimicrobial activity of amino-acids derivatives and synthetic peptides was compared to that of clinically used antibiotics. For planktonic cells, MICs of peptides and antibiotics ranged between 1 and 256 μg/ml and 256 and ≥ 2048 μg/ml, respectively. The MBCs values of Pexiganan, Citropin 1.1 and amino-acids derivatives were between 16 and 256 μg/ml, 64 and 256 μg/ml and 16 and 512 μg/ml, respectively. For clinically used antibiotics the MBCs values were above 2048 μg/ml. All of the tested peptides and amino-acids derivatives, showed inhibitory activity against P. stuartii biofilm formation, in relation to their concentrations. Pexiganan and Citropin 1.1 in concentration range 32 and 256 μg/ml caused both strong and complete suppression of biofilm formation. None of the antibiotics caused complete inhibition of biofilm formation process. The biofilm susceptibility assay verified the extremely poor antibiofilm activity of conventional antibiotics compared to synthetic peptides. The obtained results showed that synthetic peptides are generally more potent and effective than clinically used antibiotics.

Antimicrobial peptides (AMPs) are an essential part of the innate immunity of the skin and mucosal surfaces. They have a broad spectrum of antimicrobial activity: antibacterial, antifungal, antiviral as well as antiprotozoal. Numerous studies using AMPs as potential agents against different microbes has been performed during the last two decades. Here we investigated antistaphylococcal activity and safety of following AMPs: camel, citropin, protegrin, temporin A and lipopeptide Palm-KK-NH2. The susceptibility of the strains of Staphylococcus aureus isolated from the patients with erythrodermia to conventional antibiotics and AMPs was determined by the broth dilution method. The cytotoxicity assay was performed on HaCaT keratinocytes. Tested peptides turned out to be very effective against all clinical isolates, including strains resistant to conventional antibiotics. The majority of the examined peptides as well as conventional antimicrobials do not exert any toxic effect on HaCaT cells in minimal inhibitory concentration. Peptides are very promising agents for the topical treatment of staphylococcal skin infections. The most promising seem to be citropin 1.1 and temporin A, as they were toxic only in two highest concentration (50 and 100 microg/mL), with relatively low MIC values.

SUMMARY The composition of the fatty acid methyl ester (FAME) and alcohol fractions of the cuticular and internal lipids of Calliphora vomitoria larvae, pupae and male/female adults was obtained by separating these two fractions by HPLC-LLSD and analysing them quantitatively using GC-MS. Analysis of the cuticular lipids of the worldwide, medically important ectoparasite C. vomitoria revealed 6 FAMEs with odd-numbered carbon chains from C15:0 to C19:0 in the larvae, while internal lipids contained 9 FAMEs ranging from C15:1 to C19:0. Seven FAMEs from C15:0 to C19:0 were identified in the cuticular lipids of the pupae, whereas the internal lipids of the pupae contained 10 FAMEs from C13:0 to C19:0. The cuticular lipids of males and females and also the internal lipids of males contained 5, 7 and 6 FAMEs from C15:0 to C19:0 respectively. Seven FAMEs from C13:0 to C19:0 were identified in the internal lipids of females, and 7, 6, 5 and 3 alcohols were found in the cuticular lipids of larvae, pupae, males and females respectively. Only saturated alcohols with even-numbered carbon chains were present in these lipids. Only 1 alcohol (C22:0) was detected in the internal lipids of C. vomitoria larvae, while just 4 alcohols from - C18:0 to C24:0 - were identified in the internal lipids of pupae, and males and females. We also identified glycerol and cholesterol in the larvae, pupae, males and females of C. vomitoria. The individual alcohols and FAMEs, as well as their mixtures isolated from the cuticular and internal lipids of larvae, pupae, males and females of C. vomitoria, demonstrated antimicrobial activity against entomopathogenic fungi.

The cuticular and internal lipid composition in Calliphora vomitoria larvae, pupae, and male and female adults was studied. The free fatty acid (FA) compositions of the lipids were chemically characterized using gas chromatography (GC) and gas chromatography-electron impact mass spectrometry (GC-MS). Analyses of cuticular extracts from larvae, pupae, and male and female adults revealed that the carbon numbers of the acids ranged from C7:0 to C22:0, from C8:0 to C24:0, from C7:0 to C24:0 and from C7:0 to C22:0 respectively. The internal lipids of C. vomitoria larvae, pupae, male and female adults contained FAs ranging from C8:0 to C20:0, from C9:0 to C22:0, from C8:0 to C24:0 and from C9:0 to C22:0 respectively. Nine FAs with odd-numbered carbon chains from C7:0 to C21:0 were identified in the cuticular lipids of the larvae. The internal lipids of C. vomitoria larvae contained 8 odd-numbered FAs ranging from C9:0 to C19:0. Eight odd-numbered FAs from C9:0 to C21:0 were identified in the cuticular and internal lipids of pupae, while nine such FAs were found in the cuticular lipids of male and female adults. The internal lipids of adult males and females respectively contained nine and seven odd-numbered FAs, while both larvae and pupae contained eight such compounds. Eight unsaturated FAs were identified in the cuticular lipids of larvae, adult males and females and also in the internal lipids of females. Seven unsaturated FAs were identified in the cuticular lipids of pupae. The internal lipids of larvae, pupae and males contained 10, 11 and 12 unsaturated FAs respectively. Developmental changes were found both in the amounts of extracted cuticular and internal FAs and in their profiles. Four cuticular FAs (C7:0, C9:0, C10:0 and C15:1), identified as being male-specific, were either absent in the female cuticle or present there only in trace amounts. Cuticular and internal extracts obtained from larvae, pupae, adult males and females were tested for their potential antimicrobial activity. The minimal inhibitory concentrations of extracts against reference strains of bacteria and fungi were determined. Antimicrobial activity was the strongest against Gram-positive bacteria; Gram-negative bacteria, on the other hand, turned out to be resistant to all the lipids tested. Overall, the activities of the internal lipids were stronger. All the lipid extracts were equally effective against all the fungal strains examined. In contrast, crude extracts containing both cuticular and internal lipids displayed no antifungal activity against the entomopathogenic fungus Conidiobolus coronatus, which efficiently killed adult flies, but not larvae or pupae., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Information on the stimulatory and inhibitory effects of cuticular alcohols on growth and virulence of insecticidal fungi is unavailable. Therefore, we set out to describe the content of cuticular and internal alcohols in the body of housefly larvae, pupae, males and females. The total cuticular alcohols in larvae, males and females of Musca domestica were detected in comparable amounts (4.59, 3.95 and 4.03 μg g(-1) insect body, respectively), but occurred in smaller quantities in pupae (2.16 μg g(-1)). The major free alcohol in M. domestica larvae was C(12:0) (70.4%). Internal alcohols of M. domestica larvae were not found. Among cuticular pupae alcohols, C(12:0) (31.0%) was the most abundant. In the internal lipids of pupae, only five alcohols were identified in trace amounts. The most abundant alcohol in males was C(24:0) (57.5%). The percentage content of cuticular C(24:0) in males and females (57.5 and 36.5%, respectively) was significantly higher than that of cuticular lipids in larvae and pupae (0.9 and 5.6%, respectively). Only two alcohols were present in the internal lipids of males in trace amounts (C(18:0) and C(20:0)). The most abundant cuticular alcohols in females were C(24:0) (36.5%) and C(12:0) (26.8%); only two alcohols (C(18:0) and C(20:0)) were detected in comparable amounts in internal lipids (3.61±0.32 and 5.01±0.42 μg g(-1), respectively). For isolated alcohols, antimicrobial activity against 10 reference strains of bacteria and fungi was determined. Individual alcohols showed approximately equal activity against fungal strains. C(14:0) was effective against gram-positive bacteria, whereas gram-negative bacteria were resistant to all tested alcohols. Mixtures of alcohols found in cuticular lipids of larvae, pupae, males and females of M. domestica generally presented higher antimicrobial activity than individual alcohols. In contrast, crude extracts containing both cuticular and internal lipids showed no antifungal activity against the entomopathogenic fungus Conidiobolus coronatus, which efficiently kills adult house flies.

The hygroscopic secretion produced by the secretory setae of terrestrial larvae of the biting midge Forcipomyia nigra (Winnertz) was analysed using gas chromatography coupled with mass spectrometry (GC-MS). The viscous secretion is stored at the top of each seta and absorbs water from moist air. GC-MS analyses (four independent tests) showed that the secretion contained 12 free fatty acids, the most abundant of which were oleic (18:1), palmitic (16:0), palmitoleic (16:1) and linoleic (18:2). Other acids identified were valeric (5:0), enanthic (7:0), caprylic (8:0), pelargonic (9:0), capric (10:0), lauric (12:0), myristic (14:0) and stearic (18:0). Two other compounds, glycerol and pyroglutamic acid, were also found. The antibacterial activity of the fatty acids and pyroglutamic acid was tested using the agar disc diffusion method and targeted Gram positive (Bacillus cereus, Bacillus subtilis, Enterococcus faecalis) and Gram negative bacterial strains (Citrobacter freundii, Pseudomonas aeruginosa, Pseudomonas fluorescens). The antifungal activity was tested by determining minimal inhibitory concentration (MIC) of examined compounds. Fatty acids were tested against enthomopathogenic fungi (Paecilomyces lilacinus, Paecilomyces fumosoroseus, Lecanicillium lecanii, Metarhizium anisopliae, Beauveria bassiana (Tve-N39), Beauveria bassiana (Dv-1/07)). The most effective acids against bacterial and fungal growth were C(9:0), C(10:0) and C(16:1), whereas C(14:0), C(16:0,) C(18:0) and C(18:1) demonstrated rather poor antifungal activity and did not inhibit the growth of bacteria. The antimicrobial assay investigated mixtures of fatty and pyroglutamic acids (corresponding to the results of each GC-MS test): they were found to be active against almost all the bacteria except P. fluorescens and also demonstrated certain fungistatic activity against enthomopathogenic fungi. The hygroscopic secretion facilitates cuticular respiration and plays an important role in the antimicrobial protection of F. nigra larvae living in moist terrestrial habitats., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

LL37 and histatin 5 are antimicrobial peptides. LL37 exhibits killing activity against a broad spectrum of pathogens, whereas histatin 5 is primarily an antifungal agent. Head-to-tail cyclization of histatin 5 did not affect its antimicrobial and haemolytic activity. The cyclic LL37 exhibits identical antifungal and haemolytic activity as does LL37. Its antimicrobial activity varied in one dilution depending on the kind of bacteria. The structure of cyclic peptides was studied by circular dichroism spectroscopy. Both peptides undergo a conformational change leading to stabilisation of their α-helical structure in the presence of negatively charged sodium dodecyl sulfate micelles. However, with cyclic histatin 5, the presence of Zn(2+) ions is also necessary to fuse the peptide to the micelle. The specific action of the Zn(2+) ions is attributed to the presence of a zinc-binding motif, His-Glu-X-X-His. It has been speculated that this zinc complexing may be related to the well-established anticandidal activity. In the case of cyclic LL37, also the presence of a zwitterionic dodecylphosphocholine micelle induces formation of the helical structure. A microwave-assisted procedure for the cleavage of a peptide from the 2-chlorotrityl chloride resin was, for the first time, successfully used to obtain protected peptide fragments that can be applied to the preparation of head-to-tail cyclopeptides or to condensation of peptidic fragments., (Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.)