Reconstitution and expression of mcy gene cluster in the model cyanobacterium Synechococcus 7942 reveals a role of MC-LR in cell division.

Cyanobacterial blooms pose a serious threat to public health due to the presence of cyanotoxins. Microcystin-LR (MC-LR) produced by Microcystis aeruginosa is the most common cyanotoxins. Due to the limitation of isolation, purification, and genetic manipulation techniques, it is difficult to study and verify in situ the biosynthetic pathways and molecular mechanisms of MC-LR. We reassembled the biosynthetic gene cluster (mcy cluster) of MC-LR in vitro by synthetic biology, designed and constructed the strong bidirectional promoter biPpsbA2 , transformed it into Synechococcus 7942, and successfully expressed MC-LR at a level of 0.006-0.018 fg cell-1 d-1 . We found the expression of MC-LR led to abnormal cell division and cellular filamentation, further using various methods proved that by irreversibly competing its GTP-binding site, MC-LR inhibits assembly of the cell division protein FtsZ. The study represents the first reconstitution and expression of the mcy cluster and the autotrophic production of MC-LR in model cyanobacterium, which lays the foundation for resolving the microcystins biosynthesis pathway. The discovered role of MC-LR in cell division reveals a mechanism of how blooming cyanobacteria gain a competitive edge over their nonblooming counterparts.

A machine learning-based approach for low-density lipoprotein cholesterol calculation using age, and lipid parameters.

BACKGROUND: Low-density lipoprotein cholesterol (LDL-C) is a critical biomarker for cardiovascular disease. However, no consensus exists on the best method for estimating LDL-C in Chinese laboratories. This study aimed to develop a machine learning (ML) method for LDL-C estimation. METHODS: An extensive data set of 111,448 samples were randomized into five equal subsets. ML-based equations were developed using age, sex, and lipid parameters based on five-fold cross-validation. The trained ML equations were externally validated in three different data sets. The performance of the ML equations was compared with the Friedewald, Martin/Hopkins, and Sampson equations. RESULTS: The selected ML equations showed less bias with direct LDL-C than other LDL-C equations in the Chinese population, including those with triglycerides (TG) ≥ 400 mg / dL and LDL-C < 40 mg / dL. The performance of the ML equations was less susceptible to age. External validation showed the generalization of the ML equations. CONCLUSIONS: This study highlights the potential of integrating sex, age, and lipid parameters into the ML equations to obtain a more robust and reliable LDL-C calculation.

Regulation and Functional Complexity of the Chlorophyll-Binding Protein IsiA.

As the oldest known lineage of oxygen-releasing photosynthetic organisms, cyanobacteria play the key roles in helping shaping the ecology of Earth. Iron is an ideal transition metal for redox reactions in biological systems. Cyanobacteria frequently encounter iron deficiency due to the environmental oxidation of ferrous ions to ferric ions, which are highly insoluble at physiological pH. A series of responses, including architectural changes to the photosynthetic membranes, allow cyanobacteria to withstand this condition and maintain photosynthesis. Iron-stress-induced protein A (IsiA) is homologous to the cyanobacterial chlorophyll (Chl)-binding protein, photosystem II core antenna protein CP43. IsiA is the major Chl-containing protein in iron-starved cyanobacteria, binding up to 50% of the Chl in these cells, and this Chl can be released from IsiA for the reconstruction of photosystems during the recovery from iron limitation. The pigment-protein complex (CPVI-4) encoded by isiA was identified and found to be expressed under iron-deficient conditions nearly 30years ago. However, its precise function is unknown, partially due to its complex regulation; isiA expression is induced by various types of stresses and abnormal physiological states besides iron deficiency. Furthermore, IsiA forms a range of complexes that perform different functions. In this article, we describe progress in understanding the regulation and functions of IsiA based on laboratory research using model cyanobacteria.

Effects of type I collagen degradation on the durability of three adhesive systems in the early phase of dentin bonding.

OBJECTIVE: This study was designed to evaluate the effects of type I collagen degradation on the durability of three adhesive systems in the early phase of dentin bonding. METHODS: Bonded dentin specimens were prepared using three different types of adhesive systems. Micro-tensile bond strength and degradation of collagen were tested before, and after 1 month or 4 months of aging in artificial saliva. The relationship between micro-tensile bond strength and collagen degradation was analyzed by calculating their Pearson's correlation coefficient. RESULTS: Aging induced time-dependent reduction in micro-tensile bond strengths for all the tested adhesive systems, although such reduction for the single-step self-etching adhesive G-Bond (GB) was not statistically significant. The bond strength of the two-step self-etching primer adhesive system Clearfil SE Bond (SEB) was similar to that of the two-step etch-and-rinse self-priming adhesive system Single Bond 2 (SB), and they were both significantly reduced after one or four months of aging. A negative correlation was found between the degree of collagen degradation and magnitude of micro-tensile bond strength (r = -0.65, p = 0.003). The Pearson's correlation coefficient was 0.426, indicating that 42.6% of the aging-induced reduction in bond strength can be explained by the degradation of collagen. CONCLUSIONS: In the early phase of dentin bonding, there was a negative correlation between the degree of collagen degradation and the magnitude of micro-tensile bond strength. The reduction of bond strength was accompanied by the degradation of collagen. These results provide evidence for the causative relationship between the degradation of collagen and the deterioration of dentin-adhesive interface.

[Comparison of fracture resistance of pulpless teeth restored with fiber reinforced composite posts and three kinds of resin core material].

OBJECTIVE: To compare the fracture resistances of pulpless teeth restored with FRC (Fiber Reinforced Composite) posts and three kinds of resin core material. METHODS: A total of 42 recently extracted upper incisors were randomly divided into 3 groups. Group A was restored with prefabricated glass-fiber posts and Artglass polymer core; group B with prefabricated glass-fiber posts and Charisma composite resin core; and group C with prefabricated glass-fiber posts and AB composite resin core. In every group, the core material was processed by hot-press and non hot-press respectively. The posts size and shape were identical in the 3 groups. All teeth were fully covered with polycarbonate resin crowns. Fracture resistance was measured by applying point force at 130 degrees to the long axis of the teeth on an universal testing machine. RESULTS: Mean fracture threshold was 505.4 N +/- 42.0 N and 564.1 N +/- 41.7 N in group A, 411.3 N +/- 23.3 N and 315.3 N +/- 19.1 N in group B and 358.4 N +/- 36.1 N and 423.4 N +/- 47.5 N in group C. In all groups, there was no posts fracture and polycarbonate resin crowns fragmentation. CONCLUSION: The composite restoration of FRC posts combined with resin core and resin crown can improve the fracture resistance of the pulpless roots. The strength of resin core material can be increased by hot-press methods.