Tentative indicators of malaria in archaeological skeletal samples, a pilot study testing different methods.

OBJECTIVE: This study attempts to integrate multiple methods to investigate the presence of malaria in human skeletal samples from an archaeological context. MATERIALS: 33 well preserved human remains originating from a 17th-century archaeological site in southeastern Romania. METHODS: The human bone samples were analyzed using rapid diagnostic tests for malaria antigens and PCR amplification of Plasmodium falciparum apical membrane antigen 1. A preliminary test was performed to identify and briefly characterize the presence of hemozoin using a combination of TEM imaging and diffraction. RESULTS: The rapid diagnostic tests indicated that more than half of the examined samples were positive for Plasmodium antigens, but no traces of the parasites' genetic material were detected despite repeated attempts. The TEM images indicated that hemozoin might be a promising diagnostic marker of malaria in ancient bones. CONCLUSIONS: The indisputable identification of malaria in the analyzed archaeological population was not possible as none of the applied methodological strategies turned out to be straightforward. SIGNIFICANCE: This study reinforces the intricacy and limitations of unequivocally identifying malaria in past populations and sets the stage for future studies on such life-threatening infectious disease in a geographical space, which is currently underrepresented in the bioarchaeological literature. LIMITATIONS: The low sample size and the lack of consistency across all assays hindered understanding the role of malaria in the studied population. SUGGESTIONS FOR FURTHER RESEARCH: Further thorough multidisciplinary approaches on malaria detection in ancient settlements would be appropriate to inform our knowledge of its origins, frequency, and pathogen changes over centuries.

Characterization of S-layer proteins produced by lactobacilli isolated from Romanian artisan fermented products.

AIMS: To characterize S-layer proteins produced by four lactobacilli isolated from Romanian artisan fermented products. METHODS AND RESULTS: Four lactobacilli strains have been shown to produce S-layer proteins, both under optimal and stressfull conditions. The presence of S-layer proteins was confirmed by transmission electron microscopy. Removal of S-layer proteins caused a loss of the bacterial resistance to stress conditions and of the autoaggregation ability. Liquid chromatography-mass spectrometry analysis identified peptides corresponding to Slp M sequence in case of Levilactobacillus brevis 403, and peptides corresponding to Slp A sequence in case of Lactobacillus helveticus 34.9. The analysis confirmed molecular masses of ∼51 and 48 kDa, respectively, for the two proteins, and gave information about their pI, of about 9.4-9.6. A specific PCR amplification was obtained for the genome of Lact. helveticus 34.9 with slpA primers, and the amplicon sequence was 95.31% identical to slpA gene. CONCLUSIONS: Our findings indicate that certain environmental stress conditions can induce the S-layer production, which helps the producing cells to survive under unfavorable conditions.

Phyto-mediated metallic nano-architectures via Melissa officinalis L.: synthesis, characterization and biological properties.

The development of methods for obtaining new materials with antimicrobial properties, based on green chemistry principles has been a target of research over the past few years. The present paper describes the phyto-mediated synthesis of metallic nano-architectures (gold and silver) via an ethanolic extract of Melissa officinalis L. (obtained by accelerated solvent extraction). Different analytic methods were applied for the evaluation of the extract composition, as well as for the characterization of the phyto-synthesized materials. The cytogenotoxicity of the synthesized materials was evaluated by Allium cepa assay, while the antimicrobial activity was examined by applying both qualitative and quantitative methods. The results demonstrate the synthesis of silver nanoparticles (average diameter 13 nm) and gold nanoparticles (diameter of ca. 10 nm); the bi-metallic nanoparticles proved to have a core-shell flower-like structure, composed of smaller particles (ca. 8 nm). The Ag nanoparticles were found not active on nuclear DNA damage. The Au nanoparticles appeared nucleoprotective, but were aggressive in generating clastogenic aberrations in A. cepa root meristematic cells. Results of the antimicrobial assays show that silver nanoparticles were active against most of the tested strains, as the lowest MIC value being obtained against B. cereus (approx. 0.0015 mM).

S-layer production by Lactobacillus acidophilus IBB 801 under environmental stress conditions.

The ability of microorganisms to synthesize S-layer, the outermost structure of the microbial cell envelope composed of non-covalently bound proteins, has been ascribed to help microorganisms to exert their probiotic properties in the host. In this work, formation of S-layer by the potentially probiotic strain Lactobacillus acidophilus IBB 801 under different stress culture conditions (high incubation temperatures, presence of bile salts or NaCl, and acidic pH) was assayed. A marked S-layer synthesis by L. acidophilus IBB 801 was detected when the strain was grown at 42 °C and in the presence of 0.05 % bile salts or 2.0 % NaCl. The presence of S-layer proteins was further confirmed by transmission electron microscopy and protein identification by MS/MS. The differential expression of the proteome of this strain at 42 °C, when a marked formation of S-layer was detected, revealed the overexpression of six proteins mainly related to general stress and protein biosynthesis and translation, while four proteins detected in lower amounts were involved in DNA repair and energy metabolism. As L. acidophilus IBB 801 produces both a bacteriocin and S-layer proteins, the strain could be of interest to be used in the formulation of functional food products with specific properties.