Novel Natural Compounds and Their Anatomical Distribution in the Stinging Fireworm Hermodice carunculata (Annelida)

Increasing evidence in the field of bioprospection fosters the necessity of studying poorly investigated poisonous marine invertebrates to expand knowledge on animal venom biology. Among marine annelids, amphinomid fireworms are notorious for their bearded trunk equipped with a powerful stinging capacity. Here, a methodological workflow based on analytical chemistry techniques (compound isolation followed by mass spectrometry and spectroscopy analyses) was applied to gain new insights, leading to the identification and structural elucidation of an array of natural products from Mediterranean specimens of Hermodice carunculata. Eight betaine-derived unprecedented compounds, named “carunculines”, were detected, bearing two terminal ammonium groups tri-and disubstituted at the Cα (A, B) and a series of different alkyl chains (I–VIII). The mixture of chemicals was found in all the body parts of H. carunculata, supporting a mechanism of action triggered by their vehiculation inside the dorsal chaetae, and subsequent injection when chaetae break off on contact. Preliminary investigations to understand adaptive features were also performed, showing a trend in carunculine abundance that fits into the evolutionary history of these worms. These findings shed light on the chemical ecology of amphinomids, giving reasons for the success of H. carunculata in benthic environments and providing promising novel metabolites for biotechnological implications.     

Complexing the Marine Sesquiterpene Euplotin C by Means of Cyclodextrin-Based Nanosponges: A Preliminary Investigation

Euplotin C is a sesquiterpene of marine origin endowed with significant anti-microbial and anti-tumor properties. Despite the promising functional profile, its progress as a novel drug candidate has failed so far, due to its scarce solubility and poor stability in aqueous media, such as biological fluids. Therefore, overcoming these limits is an intriguing challenge for the scientific community. In this work, we synthesized β-cyclodextrin-based nanosponges and investigated their use as colloidal carriers for stably complex euplotin C. Results obtained proved the ability of the carrier to include the natural compound, showing remarkable values of both loading efficiency and capacity. Moreover, it also allowed us to preserve the chemical structure of the loaded compound, which was recovered unaltered once extracted from the complex. Therefore, the use of β-cyclodextrin-based nanosponges represents a viable option to vehiculate euplotin C, thus opening up its possible use as pharmacologically active compound.     

Computed tomography of testicular torsion in a juvenile dog with unilateral cryptorchidism

A 14-week-old male unilaterally cryptorchid Clumber spaniel was presented for acute lethargy. Physical examination revealed abdominal pain, and a single testis was palpated in the scrotum. Abdominal ultrasound and computed tomography (CT) revealed a poorly vascularized, ovoid structure immediately caudal to the left kidney with scant regional peritoneal effusion. Left intra-abdominal testicular torsion was confirmed at surgery, and routine cryptorchidectomy was performed. The patient recovered uneventfully from anesthesia and surgery.Key clinical message:The most common CT characteristics of testicular torsion were present in this case and correlated well with sonographic findings to allow for rapid, accurate diagnosis and surgical planning of unilateral, non-neoplastic, intra-abdominal cryptorchid testicular torsion in a juvenile dog. Contrast enhanced CT facilitated accurate localization of the undescended testis and evaluation of testicular perfusion and may be a useful alternative to ultrasound for diagnosing testicular torsion, especially in indeterminate cases.     

Cloning,Expression, and Purification of a GDSL-like Lipase/Acylhydrolase from a Native Lipase-Producing Bacterium,Lactobacillus fermentum

Background: Lipase enzymes are of great importance in various industries. Currently, extensive efforts have been focused on exploring new lipase producer microorganism as well as genetic and protein engineering of available lipases to improve their functional features. Methods: For screening lipase-producing lactobacilli, isolated strains were inoculated onto tributyrin agar plates. Molecular identification of lipase-producing Lactobacilli was performed by sequencing the 16Sr DNA gene, and a phylogenetic tree was constructed. The LAF_RS05195 gene, encoding lipase protein in L. fermentum isolates, was identified using specific primers, amplified by PCR (918 bp) and cloned into the pET28a (+) vector. The recombinant proteins were expressed 2, 4, 6, and 12 hours after induction with IPTG and assessed using the SDS-PAGE. Enzymatic activity of the purified recombinant protein was measured at 410 nm in the presence of ρ-NPA and ρ-NPP. Results:Among five identified native lipase-producing isolates, one isolate showed 98% similarity with Enterococcus species. The other four isolates indicated 98% similarity to L. fermentum. After purification steps with Ni-NTA column, a single protein band of about 34 kDa was detected on SDS- PAGE gel. The enzymatic activity of purified recombinant protein alongside ρ-NPA and ρ-NPP was measured to be 0.6 U/ml and 0.2 U/ml, respectively. Conclusion:In the present research, a novel lipase/esterase from L. fermentum was cloned and expressed. The novel lipase/esterase has the merit to be further studied due to its substrate specificity. Key Words: Escherichia coli, Gene expression, Lactobacillus, Lipase, Phylogeny     

Blooming phenology and self-incompatibility of some commercial cherry (Prunus avium L.) cultivars in Iran

Self-sterility in cherry (Prunus avium L.) is one of the most important problems in commercial cherry orchards in Iran and many other countries. The main objective of this research was to determine self-fertility rate and blooming phenology of 25 commercial cherry (P. avium L.) cultivars in Iran. This study was conducted during 2006–2007.Results revealed that the stage of bud swell in all cultivars began late second decade of March, which lasts 8–11 days. In all cultivars the duration end bloom to petal fall stage took 5 days. Cherry cultivars were classified into three groups considering their overlapping pollination. The cultivars each group had a great overlapped pollination toward each other. The results also showed that there was a significant difference among cultivars in respect of the fruit set (P < 1%). The fruit set of open pollination treatment among the cultivars varied in range of 7.36–41.58%. However fruit set was not observed in isolated pollination (artificial and natural self-pollination). In open pollination, the cultivars were also classified into four groups, i.e. weak fertility (Moreau and Ferracida), mean fertility (Siah Mashhad, Napelon, Proteva, Haj usefy, Lambert, Roshun, Sefid Ghermeze Baghno, Arak, Belamarka, Siah Shabestar, Ghermeze Rezaeyeh, Abardeh, Soraty Lavisan and Victoria), good fertility (Peshrase Mashhad, Dovoumras Mashhad, Bing, Shesheiy Mashhad, Siah Ghazvin and Sefid Rezaeyeh), very good fertility (Zarde Daneshkadeh, Dirrase Italia and No. 1 karaj). All cultivars were found to need cross-pollination and included in self-sterile group.     

Conserving indigenous crayfish: stock assessment and habitat requirements in the threatened Austropotamobius italicus

• 1. As part of the Austropotamobius pallipes species complex, the crayfish Austropotamobius italicus is a species of community interest whose preservation requires the designation of Special Areas of Conservation (SACs) (Annex II, EU Habitats Directive). This study aimed at (1) assessing the conservation status of this threatened indigenous species by stock assessment in central Italy and (2) identifying some aspects of its elective habitat.
• 2. Surveys were conducted in nine streams harbouring A. italicus (streams WI) and in 10 streams where crayfish populations became extinct at least 5 years before the study (streams WO).
• 3. The results confirmed that A. italicus is a K‐selected species, with a relatively slow growth rate (males: 0.34; females: 0.37) and a long life expectancy (males: 8.2 years; females: 7.8 years). The extant populations are healthy, showing balanced sex‐ratios and well structured age‐class compositions. Mortality is mainly due to fishing, which is illegal in Tuscany.
• 4. Principal Components Analyses showed that the streams WI and WO differ in the abundance of allochthonous plant detritus but not in the taxonomic composition of their macroinvertebrate communities. Age classes were found to be spatially segregated, juveniles mainly using cobbles as substrates and adults seemingly avoiding them.
• 5. The loss of the pristine riverine landscape seems to have been responsible, together with illegal fishing, for the local extinction of the species. As a consequence, retaining, enhancing, and restoring the habitat and its complexity are required for the preservation of A. italicus.
• 6. The designation of SACs might help in this endeavour if accompanied by programmes aimed at publicizing the need for conservation of this species. Unfortunately, crayfish‐focused projects supported by LIFE in Italy since 1992 (4%) and the SACs involved (1.4%) have been relatively few, despite the poor conservation status of this species and its well recognized ecological role.

Copyright © 2008 John Wiley & Sons, Ltd.