Transformation of a Methylotrophic Bacterium, Methylobacterium extorquens, with a Broad-Host-Range Plasmid by Electroporation

An efficient method for the transformation of the methylotrophic bacterium Methylobacterium extorquens NR-2 with a broad-host-range plasmid, pLA2917, by electroporation was examined. Transformants of M. extorquens NR-2 expressing resistance to kanamycin were obtained after electric pulse. These transformants were found to harbor a single plasmid which was electrophoretically identical and homologous to pLA2917 obtained from Escherichia coli. Several factors which determined the transformation efficiency were optimized, resulting in a transformation efficiency of up to 8 × 10³ transformants per μg of plasmid DNA by 10 pulses at a field strength of 10 kV/cm and a pulse duration of 300 μs.     

Characteristics of batch culture of a recombinant bacillus brevis excreting a foreign esterase

The productivity of extracellular enzyme was evaluated in batch culture using a protein hyperexcreting host, Bacillus brevis HPD31(5) harboring pHSC131, which carried a gene (est) encoding esterase activity from Bacillus stearother mophilus. Optimum temperature and pH for the bacterial growth and the production of extracellular esterase were found to be 35 degrees C and pH 6.5, by using the standard medium (GPY) containing neomycin as a selective pressure, Under the cultivation condition employed, cell growth reached 5 g dry cell weight/L, while the extracellular esterase activity amounted to 4.5 U/mL. Most (79%-92%) of the esterase produced was excreted into the medium. pHSC131 was stably retained in the host cell during cultivation in the presence of neomycin. However, in the absence of neomycin, the plasmid was completely lost from the host after 12-h cultivation accompanied by decreases in both esterase activity and production of total extracellular protein. The copy number of the plasmid was estimated to be approximately 7 throughout the cultivation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the excreted proteins showed the presence of a protein having an apparent molecular weight of 32,000, which equals to the value predicted from the DNA sequence of the est gene.     

Biochemistry and physiology of monoamine oxidase (MAO) activity in the ring dove (Streptopelia risoria). I. Characteristics of MAO activity in vitro

Monoamine oxidase (MAO) activity was measured in ring dove (Streptopelia risoria) tissues using a fluorometric assay with kynuramine as substrate. Harmaline inhibited MAO activity in a time-dependent manner, and preincubation of enzyme with the drug did not affect its activity. Pargyline produced a slow-onsetting inhibition of activity which was enhanced by preincubation of enzyme and inhibitor. Harmaline displayed reversible non-competitive inhibition of MAO activity. Oxygen is also a substrate for dove MAO, and the reaction apparently involves "ping-pong", double-displacement kinetics. Dove MAO activity is temperature-dependent, with an activation energy of 13.1 kcal/mole.     

Liquid Exfoliated Co(OH)2 Nanosheets as Low‐Cost,Yet High‐Performance,Catalysts for the Oxygen Evolution Reaction

Identifying cheap, yet effective, oxygen evolution catalysts is critical to the advancement of water splitting. Using liquid exfoliated Co(OH)₂ nanosheets as a model system, a simple procedure is developed to maximize the activity of any oxygen evolution reaction nanocatalyst. First the nanosheet edges are confirmed as the active areas by analyzing the catalytic activity as a function of nanosheet size. This allows the authors to select the smallest nanosheets (length ≈50 nm) as the best performing catalysts. While the number of active sites per unit electrode area can be increased via the electrode thickness, this is found to be impossible beyond ≈10 µm due to mechanical instabilities. However, adding carbon nanotubes increases both toughness and conductivity significantly. These enhancements mean that composite electrodes consisting of small Co(OH)₂ nanosheets and 10 wt% nanotubes can be made into freestanding films with thickness of up to 120 µm with no apparent electrical limitations. The presence of diffusion limitations results in an optimum electrode thickness of 70 µm, yielding a current density of 50 mA cm^?2 at an overpotential of 235 mV, close to the state of the art in the field. Applying this procedure to a high‐performance catalyst such as NiFeO_x should significantly surpass the state of the art.     

Origin of tendon stem cells <Emphasis Type="Italic">in situ</Emphasis>

Background

Adult stem cells are surveillance repositories capable of supplying a renewable source of progenitors for tissue repair and regeneration to maintain tissue homeostasis throughout life. Many tissue-resident stem cells have been identified in situ, which lays the foundation for studying them in their native microenvironment, i.e. the niche. Within the musculoskeletal system, muscle stem cells have been unequivocally identified in the mouse, which have led to considerable advances in understanding their role in muscle homeostasis and regeneration. On the other hand, for bone and tendon progenitor cells, mesenchymal stem cells have been used as the main in vitro cell model as they can differentiate into osteogenic, chondrogenic and tenogenic fates. Despite considerable efforts and employment of modern tools, the in vivo origins of bone and tendon stem cells remain debated. Tendon regeneration via stem cells is understudied and deserves attention as tendon damage is noted for a bleak, time-consuming recovery and the repaired tendon seldom regains the structural integrity and strength of the native, uninjured state.

Objective

Here we review the past efforts and recent studies toward defining adult tendon stem cells and understanding tendon regeneration instead of tendon development. The focus is on adult tendon resident cells in situ and the uncertainty of their roles in regeneration.

Methods

A systematic literature search using the Pubmed search engine was conducted encompassing the seminal papers in the tendon field.

Conclusion

Investigation of tendon stem cells in situ is in its infancy mainly due to lack of necessary tools and standardized injury model. We propose a concerted effort toward establishing a comprehensive cell atlas of the tendon, making genetic tools and choosing a reliable injury model for coordinated studies among different laboratories. Increasing our basic understanding should aid future therapeutic innovations to shorten and enhance the tendon repair/regeneration process.

     

Systematic studies in Neotropical <Emphasis Type="Italic">Rinorea</Emphasis> (Violaceae): Two new sections and a new generic segregate

Rinorea, the second most species-rich genus in the Violaceae, has been shown to be polyphyletic with four separate clades recovered in phylogenetic studies. Among these clades is the Rinorea crenata group, which is composed of three Neotropical species. This group has been shown in family- and genus-level molecular phylogenies to be resolved outside of a large clade representing Rinorea s.str. Based on phylogenetic, morphological, and anatomical evidence, Bribria, a new genus, is segregated from Rinorea s.str. and described, with new combinations made for its three species: Bribria apiculata, Bribria crenata, and Bribria oraria. In addition, two new sections in Rinorea s.str. are described to accommodate the remaining Neotropical species: Rinorea sect. Rinorea and Rinorea sect. Pubiflora, which correspond to Group IIa Rinorea and Group IIc Pubiflora, respectively, in W. H. A. Hekking’s monograph of Neotropical Rinorea.     

Phenetic and cladistic studies help clarify species assemblages in <Emphasis Type="Italic">Passiflora</Emphasis> section <Emphasis Type="Italic">Dysosmia</Emphasis> (Passifloraceae)

One of the most enigmatic and unusual groups in the passionflower genus, Passiflora section Dysosmia (Passifloraceae), stands out as a group that is notoriously taxonomically complicated. Phenetic and cladistic analyses of Dysosmia were carried out with representatives from all 21 species currently recognized in the section, in order to delineate morphological groups above the species level. The study was based mainly on vegetative morphological characters and included principal coordinates, non-metric multidimensional scaling, cluster, and cladistic analyses. Results from each analysis reveals that three major species assemblages exist within Dysosmia, corresponding largely to vegetative pubescence and fruit color. The results presented here set the stage for future systematic and phylogenetic studies in Passiflora sect. Dysosmia.     

Small carbonaceous fossils (SCFs) from the Terreneuvian (lower Cambrian) of Baltica

We describe a new assemblage of small carbonaceous fossils (SCFs) from diagenetically minimally altered clays and siltstones of Terreneuvian age from the Lontova and Voosi formations of Estonia, Lithuania and Russia. This is the first detailed account of an SCF assemblage from the Terreneuvian and includes a number of previously undocumented Cambrian organisms. Recognizably bilaterian‐derived SCFs include abundant protoconodonts (total‐group Chaetognatha), and distinctive cuticular spines of scalidophoran worms. Alongside these metazoan remains are a range of protistan‐grade fossils, including Retiranus balticus gen. et sp. nov., a distinctive funnel‐shaped or sheet‐like problematicum characterized by terminal or marginal vesicles, and Lontohystrichosphaera grandis gen. et sp. nov., a large (100–550 μm) ornamented vesicular microfossil. Together these data offer a fundamentally enriched view of Terreneuvian life in the epicratonic seas of Baltica, from an episode where records of non‐biomineralized life are currently sparse. Even so, the recovered assemblages contain a lower diversity of metazoans than SCF biotas from younger (Stage 4) Baltic successions that represent broadly equivalent environments, echoing the diversification signal recorded in the coeval shelly and trace‐fossil records. Close comparison to the biostratigraphical signal from Fortunian small shelly fossils supports a late Fortunian age for most of the Lontova/Voosi succession, rather than a younger (wholly Stage 2) range.     

A Docosahexaenoic Acid-Derived Pro-resolving Agent,Maresin 1, Protects Motor Neuron Cells Death

Maresin 1 is a novel pro-resolving mediator derived from docosahexaenoic acid (DHA), with potent anti-inflammation effects against several animal models, including brain ischemia, sepsis, and lung fibrosis. However, its effect against motor neuron cell death is still not investigated. Therefore, we investigated the effects of maresin 1 on several stress-induced motor neuron cell death. Maresin 1 suppressed combinatorial stress which was evoked by superoxide dismutase 1 (SOD1)^G93A and serum-free, -induced motor neuron cells death in a concentration-dependent manner, and had a stronger neuroprotective effective than DHA. Maresin 1 also had neuroprotective effects against transactivation response DNA-binding protein (TDP)-43^A315T and serum-free stress, H₂O₂, and tunicamycin-induced cell death. Maresin 1 reduced the reactive oxygen species (ROS) production caused by SOD1^G93A or TDP-43^A315T. Moreover, maresin 1 suppressed the NF-κB activation induced by SOD1^G93A and serum-free stress. These data indicate that maresin 1 has motor neuron protective effects against several stresses by reduction of ROS production or attenuation of the NF-κB activation. Maresin 1 also had neuroprotective effects against H₂O₂, and tunicamycin-induced cell death in a concentration-dependent manner. Finally, maresin 1 ameliorated the motor function deficits of spinal muscular atrophy model in which endoplasmic reticulum stress was upregulated. Thus, maresin 1 may be beneficial to protect against motor neuron diseases.     

Protein-restricted maternal diet during lactation decreases type I and type III tropocollagen synthesis in the skin of mice offspring

We investigated the effects of a low protein (LP) maternal diet during lactation on type I and III tropocollagen synthesis in infant mouse skin. The LP diet decreased the levels of type I and III tropocollagen proteins and COL1A1 and COL3A1 mRNA. Thus, the protein composition of the maternal perinatal diet may influence the skin health of offspring.