Large net cage for captive breeding and behavioural studies of damselfly Lestes sponsa (Hansemann, 1823) (Odonata: Lestidae): submerged oviposition as a model behaviour

Odonata have a strong potential as model organisms for testing ecological and evolutionary hypotheses because of their short life history, relative ease and cost-effectiveness of care. Unfortunately, very few studies have examined how to create a semi-natural environment for odonates, limiting the biological validity of laboratory manipulation. To better study odonate life cycle and behaviour under controlled conditions, we designed a large net cage that imitated the natural terrestrial as well as aquatic habitat of the damselfly Lestes sponsa (Hansemann, 1823). This species is thought to be capable of submerged oviposition, an unusual behaviour in odonates. We compared multiple variables across natural conditions and the net cage. We demonstrated that between-year variability under natural conditions was generally greater than variability between natural and artificial environments. Overall, semi-natural conditions did not substantially change the L. sponsa life cycle (including the unique behaviour of submerged oviposition), suggesting that results from the net cage are likely generalisable to the field.     

Technical comment on Condamine et al. (2019): a cautionary note for users of linear diversification dependencies

Condamine et al. (2019; 22: 1900–1912) fitted linear and exponential functions of time‐dependent, diversity‐dependent and temperature‐dependent diversification to investigate diversification dynamics of tetrapod families. Here I highlight potential misinterpretations when using linear diversification dependencies and provide some clarifications.     

Technical note: out-of-plane angular correction based on a trigonometric function for use in two-dimensional kinematic studies

In two-dimensional (2D) kinematic studies, limb positions in three-dimensional (3D) space observed in lateral view are projected onto a 2D film plane. Elbow and knee-joint angles that are less than 20 degrees out-of-plane of lateral-view cameras generally exhibit very little measurable difference from their 3D counterparts (Plagenhoef 1979 Environment, Behavior, and Morphology; New York: Gustav Fisher, p. 95-118). However, when limb segment angles are more than 20 degrees out-of-plane, as is often the case in locomotor studies of arboreal primates, elbow and knee angles can appear significantly more extended than they actually are. For this reason, a methodology is described that corrects 2D out-of-plane angular estimates using a series of trigonometric transformations.     

Chemical modification of the small intestinal Na+/d-glucose cotransporter by amino group reagents: Evidence for a role of amino group(s) in the binding of the sugar

Some amino group reagents inactivate the small-intestinal Na⁺/d-glucose cotransporter, as measured either as a catalyst of Na⁺-dependent d-glucose transport or as a Na⁺-dependent phlorizin ligand. The amino group(s) studied in this paper are not identical with those investigated previously (Biber, J., Weber, J. and Semenza, G. (1983) Biochim. Biochim. Acta 728, 429–437): these are protected from inactivation by the simultaneous presence of Na⁺ plus sugar substrates. They are thus likely to be located within the substrate-binding site.     

Mechanistic studies on carboxypeptidase a from goat pancreas — part II: Evidence for carboxyl group

Studies with substrate analogues and the pH optimum indicated the involvement of carboxyl group in the active site of goat carboxypeptidase A. Chemical modification of the enzyme with 1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide methoI -p-toluene sulphonate, a carboxyl specific reagent, led to loss of both esterase and peptidase activities. Protection studies showed that this carboxyl group was in the active site and was protected by Βp-phenylpropionic acid and glycyl-L-tyrosine. Kinetic studies also confirmed the involvement of carboxylic group because the enzyme modification with water soluble carbodiimide was a two step reaction which excluded the possibility of tyrosine or lysine which are known to give a one step reaction with this reagent     

Coffee husk: an inexpensive substrate for production of citric acid by Aspergillus niger in a solid-state fermentation system

Aspergillus niger CFTRI 30 produced 1.3 g citric acid/10 g dry coffee husk in 72 h solid-state fermentation when the substrate was moistened with 0.075 M NaOH solution. Production was increased by 17% by adding a mixture of iron, copper and zinc to the medium but enrichment of the moist solid medium with (NH₄)₂SO₄, sucrose or any of four enzymes did not improve production. The production of about 1.5 g citric acid/10 g dry coffee husk at a conversion of 82% (based on sugar consumed) under standardized conditions demonstrates the commercial potential of using the husk in this way.The authors are with the Department of Microbiology and Bioengineering, Central Food Technological Research Institute, Mysore-570 013, India;     

Positional behavior and body size of arboreal primates: a theoretical framework for field studies and an illustration of its application.

The rationale for most field studies of the positional behavior of arboreal primates has been the need to document natural behaviors quantitatively in order to infer the functional significance of morphological configurations. This focus on interactions of morphology with behavior is justifiable, but there exists another important level of biological relationships, that of the animal with its structural habitat, which it must negotiate to find food and avoid being preyed on. Recently it has become apparent that body size is likely to affect relationships of positional behavior with habitat structure, as well as with morphology. Here I offer a framework for research on functional relationships of positional behavior, body size, and habitat structure, with the ultimate objective of elucidating the aptive significance of the great diversity exhibited by arboreal primates. This approach specifies several distinct problems that animals solve, and indicates how research might be directed at revealing the relative effectiveness with which different primates solve them. A preliminary application of the framework examines sympatric north Sumatran primates.     

Technical note: GAMMORA,a free,open-source,and validated GATE-based model for Monte-Carlo simulations of the Varian TrueBeam

PurposeMonte Carlo (MC) is the reference computation method for medical physics. In radiotherapy, MC computations are necessary for some issues (such as assessing figures of merit, double checks, and dose conversions). A tool based on GATE is proposed to easily create full MC simulations of the Varian TrueBeam STx.MethodsGAMMORA is a package that contains photon phase spaces as a pre-trained generative adversarial network (GAN) and the TrueBeam’s full geometry. It allows users to easily create MC simulations for simple or complex radiotherapy plans such as VMAT. To validate the model, the characteristics of generated photons are first compared to those provided by Varian (IAEA format). Simulated data are also compared to measurements in water and heterogeneous media. Simulations of 8 SBRT plans are compared to measurements (in a phantom). Two examples of applications (a second check and interplay effect assessment) are presented.ResultsThe simulated photons generated by the GAN have the same characteristics (energy, position, and direction) as the IAEA data. Computed dose distributions of simple cases (in water) and complex plans delivered in a phantom are compared to measurements, and the Gamma index (3%/3mm) was always superior to 98%. The feasibility of both clinical applications is shown.ConclusionsThis model is now shared as a free and open-source tool that generates radiotherapy MC simulations. It has been validated and used for five years. Several applications can be envisaged for research and clinical purposes.     

Technical note: Hapten synthesis,antibody production and development of an enzyme-linked immunosorbent assay for detection of the natural steroidal alkaloid Dendrogenin A

We have recently discovered the existence of 5α-Hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-ol, called Dendrogenin A (DDA), as the first endogenous steroidal alkaloid ever described in mammals. We found that the DDA content of tumors and cancer cell lines was low or absent compared with normal cells showing that a deregulation in DDA biosynthesis was associated with cancer and therefore suggesting that DDA could represent a metabolomic cancer biomarker. This prompted us to produce antibodies that selectively recognize DDA. For this purpose, the hapten 5α-hydroxy-6β-[2-(1H-imidazol-4-yl)ethylamino]cholestan-3β-o-hemisuccinate with a carboxylic spacer arm attached to the 3β-hydroxyl group of DDA was synthesized. The hapten was coupled to bovine serum albumin and keyhole limpet hemocyanin for antibody production to develop an enzyme-linked immunosorbent assay (ELISA). The protein conjugates were injected into BALB/c mice to raise antibodies. The monoclonal antibodies that were secreted from the hybridoma cell lines established were assessed with indirect ELISA by competitive assays using dilutions of a DDA standard. The antibodies from the selected hybridomas had an IC₅₀ value ranging from 0.8 to 425 ng/ml. Three antibodies showed no cross-reactivity with structurally related compounds including histamine, cholesterol, ring B oxysterols and a regio-isomer of DDA. In this study, high-affinity and selective antibodies against DDA were produced for the first time, and a competitive indirect ELISA was developed.     

Rhesus macaque (Macaca mulatta) group formation and housing: wounding and reproduction in a specific pathogen free (SPF) colony.

In the present report, we examined the effects of group formation strategy and corral design on wounding and reproduction rates in rhesus macaques. Specifically, we examined group formation using a staged strategy, in which small groups of animals were introduced incrementally over a period of weeks, and a rapid formation strategy, in which all animals were introduced in 1 day. We also examined group formation using a divided corral design that facilitated visual and social separation of individuals, and an undivided corral design that did not facilitate visual or social separation. Dependent measures were wounding and reproductive rates over each of the 2 years that followed group formation. Results indicate that incrementally releasing subgroups of animals, and using a corral design that provides for visual and social separation of individuals, are effective strategies for reducing rates of traumatic wounding when forming multimale-multifemale rhesus macaque breeding groups. However, it must be noted that differences in formation strategy and corral design did not lead to higher reproductive rates. We conclude that incrementally releasing animals in hierarchical subgroups, and using a divided vs. undivided housing design, reduced intra-group wounding and associated demands on veterinary and animal management resources following formation of rhesus macaque breeding groups.     

Comparison of three different farrowing systems: skin lesions and behaviour of sows with special regard to nursing behaviour in a group housing system for lactating sows

While group housing (GH) is mandatory in the European Union for the greater part of pregnancy, single housing in farrowing crates (FCs) during lactation that restrict sows in most of their natural behaviour patterns is still practised on a large scale. Research is urgently needed to develop alternative farrowing systems that improve sows’ welfare. Therefore, sows in three different farrowing systems – pens with FC, loose housing (LH) pens and GH for six sows – were compared regarding the level of skin injuries and their active and resting behaviour. A skin injury score was assessed for 15 body parts of 102 sows in six batches on 3 days (days 1, 14 and 34). In total, the active and resting behaviour of 77 sows in six batches was examined on 3 days (days 18, 25 and 32) between 0700 h and 1900 h by means of a scan sampling method. The suckling behaviour and the level of cross-suckling were analysed in GH by means of direct observation in four batches during three 4-h sampling periods (days 17, 24 and 31). No significant differences were found in total skin injuries when the sows entered the systems (day 1), but GH sows showed significantly higher total skin injuries compared to FC and LH sows in the middle (day 14) and at the end (day 34) of the lactation period. A significant difference between FC and LH sows was never seen. Differences were found for the proportion of different body postures between the three systems. The odds for lying in lateral recumbency versus standing and sitting versus standing were significantly higher for FC and LH sows compared to GH sows. Additionally, sows were significantly more likely to be standing as opposed to lying in lateral recumbency as the lactation period progressed. Cross-suckling was a frequent behaviour in GH, seen in 35.0% of all successful suckling bouts. However, only an average of 0.56 piglets per successful suckling bout was observed cross-suckling, suggesting only a few piglets were engaged in cross-suckling. In conclusion, the skin injury score was only moderately increased in GH compared to FC and LH and comparable to pregnant group-housed sows, both free farrowing systems seemed to be an environmental enrichment for lactating sows and good management cannot prevent the occurrence of cross-suckling in a GH system, but can probably reduce it.     

Synthesis and characterization of a novel reagent containing dansyl group, which specifically alkylates sulfhydryl group: an example of application for protein chemistry

We have synthesized a novel reagent containing dansyl group, iodoacethyl dansylcadaverine (IADC), which specifically alkylates sulfhydryl groups. The carboxyl group of iodoacetic acid was activated with dicyclohexylcarbodiimide and was condensed with amino group of dansylcadaverine. Purity and chemical structure of IADC was confirmed with mass spectrometry (MS) and NMR. IADC alkylated GSH but not GSSG, which was confirmed by MS. The reactivity of IADC with proteins was also investigated with Western blotting using anti-dansyl antibody. IADC reacted only with sulfhydryl-containing proteins. The specificity of the interaction of IADC with sulfhydryl groups in proteins was confirmed by adding excessive amount of a well-known sulfhydryl-specific reagent, 5, 5'-dithiobis(2-nitrobenzoic acid), which led to a complete inhibition. To show the usefulness of IADC, the cysteines in glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from chicken muscle were modified with this reagent, and GAPDH was then digested by lysyl endopeptidase. The peptides generated from digestion of IADC-incorporated GAPDH were applied to an anti-dansyl immunoaffinity column. The peptide fragments bound and eluted from the column were separated by HPLC, and the amino acid sequence of each peptide was analyzed, and peptide was identified as the one containing a Cys residue(s). These data showed that IADC is a useful reagent to specifically identify the positions of a Cys residue(s) in proteins.     

Staphostatins: an expanding new group of proteinase inhibitors with a unique specificity for the regulation of staphopains,Staphylococcus spp. cysteine proteinases

A novel type of cysteine proteinase inhibitor (SspC) has been recently recognized in Staphylococcus aureus (Massimi, I., Park, E., Rice, K., Muller-Esterl, W., Sauder, D.N., and McGavin, M.J. (2002) J Biol Chem 277: 41770-41777). In this paper we have identified homologous proteins encoded in the genome of S. aureus and other coagulase-negative Staphylococci. Collectively we refer to these proteins as staphostatins as they specifically inhibit cysteine proteinases (staphopains) from Staphylococcus spp. The primary structure of staphostatins seems to be unique, although they resemble cystatins in size (105-108 residues). Recombinant staphostatin A, a product of the scpB gene and staphostatin B (SspC) from S. aureus have been characterized in details. Similar to the cystatins, the staphostatins interact specifically with their target proteinases forming tight and stable non-covalent complexes, staphostatin A with staphopain A and staphostatin B with staphopain B. However, in contrast to the cystatins, each of which inhibits broad range of cathepsins, complex formation between staphostatin and staphopain appears to be exclusive, with no cross interaction observed. In addition, the activities of several tested cysteine proteinases of prokaryotic- and eukaryotic-origin were not affected by staphostatins. Such narrow specificity limited to staphopains is presumed to be required to protect staphylococcal cytoplasmic proteins from being degraded by prematurely activated/folded prostaphopains. This function is guaranteed through the unique co-expression of the secreted proteinase and the intracellular inhibitor from the same operon, and represents a unique mechanism of regulation of proteolytic activity in Gram-positive bacteria.