Allantoin from Valuable Romanian Animal and Plant Sources with Promising Anti-Inflammatory Activity as a Nutricosmetic Ingredient

Helix aspersa (HA), Helix pomatia (HP) and Symphytum officinale are common organisms in Romania’s biosphere, widely known for their allantoin content and their therapeutic properties. Herein, the allantoin was separated and quantified from the aqueous extracts of Romanian comfrey root and the secretions of HA and HP snails. This study also focused on determining the antioxidant and anti-inflammatory activities of these Romanian allantoin-rich samples. The plant extracts were obtained through two methods: ultrasonic extraction and enzymatic ultrasonic extraction. A microplate method was used for the quantitative determination of allantoin content. The antioxidant activity was measured by using the DPPH radical scavenging method. The antioxidant capacity of the samples was studied in order to observe the type of interactions generated by the chemical complex present in their composition. High concentrations of allantoin were obtained by enzymatic ultrasonic extraction method (EUE—102 ± 0.74 μg/mL), and also in the water-soluble fraction of the snail secretion (FS1—22.051 μg/mL). The antioxidant screening suggests that Symphytum officinale and snail mucus extracts could be used as promising natural substitutes for synthetic antioxidants in products used for therapeutic purposes. The evaluation of anti-inflammatory activity was also investigated, allantoin-rich samples showing a promising action (FS1—81.87 ± 2.34%). In future, the inclusion of allantoin-rich extracts in various novel pharmaceutical forms for new therapeutic applications could be achieved. The study will continue with the formulation of a nutricosmetic product with snail mucus and Symphytum officinale extract as principal bioactive ingredients.

A comparative genetic study on exploited vs. Unperturbed wild populations of Helix Pomatia (L., 1758): preliminary results

Helix pomatia is one of the most well known species of land snails across Europe and it presents economical importance due to its consumption as food. The preliminary results of the genetic comparison between two populations, one of them under the pressure of exploitation, are presented here. As expected, most indices revealed a disequilibrium in the exploited population, with the exeption of the allelic pattern which was similar among the two studied populations.

The T/Tn-Specific Helix pomatia Lectin Induces Cell Death in Lymphoma Cells Negative for T/Tn Antigens

Morniga G is a T/Tn-specific lectin, inducing cell death in Tn-positive leukemias but not in healthy lymphocytes. Helix pomatia lectin (HPA) is another T/Tn-specific lectin, currently used as tool for cancer diagnostics. The HPA-mediated tumor cell death was evaluated on human leukemia and mouse lymphoma cells, and compared to the effect of Morniga G. Both lectins induced an equivalent percentage of cell death in Tn-positive Jurkat human leukemia. In contrast, EL4 mouse lymphoma resisted Morniga G-mediated cytotoxicity but were killed by HPA at concentrations of 2.5 μg/mL (0.032 nM) and higher. In both malignant cells, HPA-mediated cell death showed features compatible with apoptosis (annexin-externalization, caspase-activation, mitochondrial membrane depolarization, and ROS production). Cytometry analysis indicated that EL4 cells are T/Tn-negative. Because previous results showed a high amount of N-acetylgalactosamine (GalNAc, sugar present in Tn antigen) on EL4 cell surface, this GalNAc could be involved in the formation of truncated O-glycans other than the T/Tn residues. When compared to Morniga G, bioinformatic analysis suggested that HPA benefits from an extended carbohydrate-binding site, better adapted than Morniga G to the accommodation of more complex branched and truncated O-glycans (such as core 2). Finally, HPA killed EL4 cells but not healthy lymphocytes in a mixture of lymphoma cells + lymphocytes, suggesting that HPA selectively triggers tumor cell death.

Experimental indications of gardeners’ anecdotes that snails interfere with invasive slugs

The invasive Spanish slug (Arion vulgaris) is an important pest species in agriculture and horticulture in Europe. In the last decades it has spread across the continent where it outcompetes native slug and snail species, thus posing a threat for biodiversity. A popular anecdote suggests to promote Roman snails (Helix pomatia) in gardens because they are able to control A. vulgaris. We examined a potential interrelationship between these two species using a mesocosm experiment with lettuce plants. 13C-15N stable isotope labelling of lettuce allowed us to investigate interactions between Helix and Arion on weight gain/loss and herbivory. Additionally, we wanted to know whether different watering regimes (daily vs. every 3rd day watering of weekly amount) and earthworms alter these interactions. Egg predation of Helix on Arion eggs was further tested in a food-choice experiment. Arion showed a five times higher herbivory per body mass than Helix in a single-species setting. However, in mesocosms containing both species percentage of herbivory per body mass was significantly lower than in Arion-only mesocosms, especially when watered every three days. Overall isotope uptake via eaten lettuce was unaffected by the presence of the other species. Only very little predation (three out of 200 eggs) of Helix on Arion eggs was observed. Our results provide no evidence for a clear dismissal or confirmation of the popular gardener’s anecdote that Helix snails have a negative effect on Arion abundance or herbivory.

Analysis of the adventitious fauna of terrestrial molluscs in the centre of the Russian Plain

The terrestrial mollusc fauna of the central part of the Russian Plain has been analysed with respect to its species composition and the proportion of adventitious species. The classification of adventitious species follows that accepted in Russia. Considerable changes in the fauna took place in the period 2016–2020. Helix lucorum Linnaeus and Cornu aspersum (O. F. Müller) were added to the terrestrial malacofauna of the area. Within the last four years, Arion vulgaris Moquin-Tandon colonised anthropogenic habitats and its spread continues. Krynickillus melanocephalus Kaleniczenko, Arianta arbustorum (Linnaeus), and Helix pomatia Linnaeus are now firmly established in natural ecosystems. Populations of A. vulgaris, K. melanocephalus, and A. arbustorum have become so abundant that now they cause significant damage to agricultural crops. In both natural and anthropogenic habitats K. melanocephalus and A. arbustorum displace Succinea putris (Linnaeus), Deroceras laeve (O. F. Müller), D. sturanyi (Simroth), D. invadens Reise, Hutchinson, Schunack et Schlitt, 2011, D. agreste (Linnaeus), D. reticulatum (O. F. Müller), Fruticicola fruticum (O. F. Müller) and Euomphalia strigella (Draparnaud). Interactions between invasive species are discussed. The proportion of alien species in several European countries and in Central Asia is compared.

The evolution of hemocyanin genes in Tectipleura: a multitude of conserved introns in highly diverse gastropods

Background Hemocyanin is the oxygen transporter of most molluscs. Since the oxygen affinity of hemocyanin is strongly temperature-dependent, this essential protein needs to be well-adapted to the environment. In Tectipleura, a very diverse group of gastropods with > 27,000 species living in all kinds of habitats, several hemocyanin genes have already been analyzed. Multiple independent duplications of this gene have been identified and may represent potential adaptations to different environments and lifestyles. The aim of this study is to further explore the evolution of these genes by analyzing their exon–intron architectures. Results We have reconstructed the gene architectures of ten hemocyanin genes from four Tectipleura species: Aplysia californica, Lymnaea stagnalis, Cornu aspersum and Helix pomatia. Their hemocyanin genes each contain 53 introns, significantly more than in the hemocyanin genes of Cephalopoda (9–11), Vetigastropoda (15) and Caenogastropoda (28–33). The gene structures of Tectipleura hemocyanins are identical in terms of intron number and location, with the exception of one out of two hemocyanin genes of L. stagnalis that comprises one additional intron. We found that gene structures that differ between molluscan lineages most probably evolved more recently through independent intron gains. Conclusions The strict conservation of the large number of introns in Tectipleura hemocyanin genes over 200 million years suggests the influence of a selective pressure on this gene structure. While we could not identify conserved sequence motifs within these introns, it may be simply the great number of introns that offers increased possibilities of gene regulation relative to hemocyanin genes with less introns and thus may have facilitated habitat shifts and speciation events. This hypothesis is supported by the relatively high number of introns within the hemocyanin genes of Pomacea canaliculata that has evolved independently of the Tectipleura. Pomacea canaliculata belongs to the Caenogastropoda, the sister group of Heterobranchia (that encompass Tectipleura) which is also very diverse and comprises species living in different habitats. Our findings provide a hint to some of the molecular mechanisms that may have supported the spectacular radiation of one of Metazoa’s most species rich groups.

ПЕРСПЕКТИВА ВИКОРИСТАННЯ СЛИЗУ РІЗНИХ ВИДІВ РАВЛИКІВ У МЕДИЦИНІ

Резюме. Здавна відомо про терапевтичні властивості слизу різних видів равликів. У світі зростає інтерес науковців щодо можливостей застосування слизу равликів різних видів, зокрема Helix pomatia, Helix aspersa maxima, Helix aspersa muller у медицині. Мета дослідження – узагальнити дані літературних джерел щодо перспективності використання слизу равликів як активного фармацевтичного інгредієнта у медичній галузі та у фармацевтичній промисловості. Матеріали і методи. Проаналізовано та узагальнено дані понад 50 джерел, із яких у даному огляді використано 30 джерел сучасної іноземної та вітчизняної літератури щодо доцільності використання слизу равликів у медицині, фармації та косметології. У дослідженні опрацьовано наукові публікації за останнє десятиліття, які доступні у мережі «Інтернет», ключовими словами були муцин, слиз, равлик, ліофілізація, технологія ліків. Результати. На основі представлених літературних даних можна зробити висновок, що слиз равлика має велике практичне значення для різних галузей медицини (дерматологія, ортопедія, терапія), широко використовується у технології лікарських засобів як активний фармацевтичний інгредієнт та основа для трансдермальних терапевтичних систем. Фармакологічна активність слизу равликів забезпечується збагаченим хімічним складом, адже активними компонентами є протеоглікани, глікозаміноглікани (зокрема, гіалуронова кислота), глікопротеїнові ферменти, амінокислоти, пептиди (мідні, антимікробні), вітаміни (A, C, E, B1 та B6), макро- та мікроелементи (Na, Ca, K, Mg, P, Cu, Zn, Fe, Mn), муцин, колаген, гліколева кислота, еластин та алантоїн. Доведено, що слиз равликів має антиоксидантні, антибактеріальні, регенеративні, протизапальні властивості, а також при його застосуванні забезпечується зволожувальна, очищувальна та захисна дії. Висновки. На сьогодні слиз равликів широко використовують у дерматології та косметології, що підтверджують результати фармакологічних досліджень. Актуальним та перспективним є використання слизу як основи-носія для створення лікарських засобів, а також як активного фармацевтичного інгредієнта з широким спектром фармакологічної дії.