Diversity and Bioprospective Potential (Cold-Active Enzymes) of Cultivable Marine Bacteria from the Subarctic Glacial Fjord, Kongsfjorden

The diversity and abundance of culturable bacteria in Kongsfjorden water (15 stations) and sediments (12 stations) were studied. Viable numbers ranged between 10⁵–10⁶ CFU l^?1 in water and 10²–10⁴ CFU g^?1 in the sediments. A total of 291 and 43 bacterial isolates were retrieved from the water (KJF) and sediments (FS), respectively. Based on 16S rRNA gene sequence similarities, the KJF and FS isolates were grouped into 49 and 23 phylotypes, respectively. The KJF and FS phylotypes represented three phyla namely, Actinobacteria, Bacteroidetes, and Proteobacteria. At the genus level, Flavobacterium and Shewanella and at the species level, Pseudoaltermonas arctica and Colwellia psychrerythraea were dominant in the water and sediments, respectively. Most phylotypes were psychrotolerant with upper growth temperature limit of 25–37 °C and tolerated 0.3–2.5 M NaCl and pH values of 5.0–11.0. Majority of the phylotypes produced one or more of the extracellular hydrolytic enzymes amylase, lipase, caseinase, urease, gelatinase, and DNase at 4 and 18 °C, while none were chitinolytic. Few of the FS phylotypes exhibited extracellular activity only at 4 or 18 °C. Nine FS and 21 KJF isolates were pigmented. The predominant cellular fatty acids were unsaturated, branched, and modified fatty acids, which are unique to cold-adapted bacteria.     

Methanopyrus kandleri topoisomerase V contains three distinct AP lyase active sites in addition to the topoisomerase active site

Topoisomerase V (Topo-V) is the only topoisomerase with both topoisomerase and DNA repair activities. The topoisomerase activity is conferred by a small alpha-helical domain, whereas the AP lyase activity is found in a region formed by 12 tandem helix-hairpin-helix ((HhH)₂) domains. Although it was known that Topo-V has multiple repair sites, only one had been mapped. Here, we show that Topo-V has three AP lyase sites. The atomic structure and Small Angle X-ray Scattering studies of a 97 kDa fragment spanning the topoisomerase and 10 (HhH)₂ domains reveal that the (HhH)₂ domains extend away from the topoisomerase domain. A combination of biochemical and structural observations allow the mapping of the second repair site to the junction of the 9th and 10th (HhH)₂ domains. The second site is structurally similar to the first one and to the sites found in other AP lyases. The 3rd AP lyase site is located in the 12th (HhH)₂ domain. The results show that Topo-V is an unusual protein: it is the only known protein with more than one (HhH)₂ domain, the only known topoisomerase with dual activities and is also unique by having three AP lyase repair sites in the same polypeptide.     

Pathogenic LMNA variants disrupt cardiac lamina-chromatin interactions and de-repress alternative fate genes

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Preclinical pharmacokinetic characterization of an adipose tissue-targeting monoclonal antibody in obese and non-obese animals

Target receptor levels can influence pharmacokinetics (PK) or pharmacodynamics (PD) of monoclonal antibodies (mAbs), and can affect drug development of this class of molecules. We generated an effector-less humanized bispecific antibody that selectively activates fibroblast growth factor receptor (FGFR)1 and βKlotho receptor, a FGF21 receptor complex highly expressed in both white and brown adipocytes. The molecule shows cross-species binding with comparable equilibrium binding affinity (Kd) for human, cynomolgus monkey, and mouse FGFR1/βKlotho. To understand the PK/PD relationship in non-obese and obese animals, we evaluated the adipose tissue distribution of the antibody, serum exposures, and an associated PD marker (high-molecular-weight adiponectin), in both non-obese and obese mice and monkeys. Antibody uptake into fat tissue was found to be higher on a per gram basis in non-obese animals compared to obese animals. Since obesity has been reported to be associated with reduced expression of FGFR1 and βKlotho receptor in white adipose tissues in mice, our results suggest that the distribution in adipose tissues was influenced by target expression levels. Even so, the overall dose-normalized serum exposures were comparable between non-obese and obese mice and monkeys, suggesting that adipose tissue uptake plays a limited role in overall systemic PK determination. It remains to be determined if and how obesity and receptor expression in humans influence the PK and PD profile of this novel therapeutic candidate.     

Distribution of metal ions in the subcellular fractions of several rat brain areas.

The levels of Cu, Fe, Zn, Mg and Ca in the synaptosomal myelin and mitochondrial fractions of the rat brain were determined quantitatively by means of the atomic absorption spectroscopic method. In the whole brain the concentrations of the metals were present in the three different fractions in the following range; Cu, 0.46-1.76; Fe, 0.24-3.51; Zn, 0.12-0.93; Mg, 1.08-1.41 and Ca, 1.28-3.56 μg/mg tissue protein. Analyses of the subcellular organelles prepared from different areas of the brain indicated that the concentrations of Cu and Zn were relatively larger in the hypothalamus and lower in cerebellum in comparison with other areas. On treatment of the rats with the metal chelating agents i.e., EDA, EDDHA and HBED intracisternally, it was found that the distribution of the subcellular metals was markedly affected.