Flocculation and pentadecane production of a novel filamentous cyanobacterium <Emphasis Type="Italic">Limnothrix</Emphasis> sp. strain SK1-2-1

Objective

A novel filamentous cyanobacterium, a photosynthesizing microorganism, was isolated from a river, and its unique features of flocculation and pentadecane production were characterized.

Results

Microscopic observations and a phylogenetic analysis with 16S rDNA revealed that this strain was a Limnothrix species denoted as the SK1-2-1 strain. Auto cell-flocculation was observed when this strain was exposed to a two-step incubation involving a standing cultivation following a shaking preincubation. Flocculation was enhanced by blue light at a wavelength at 470 nm and irradiation for several hours to 1 day. Moreover, the strain exhibiting exponential cell growth may preferentially accumulate alkanes as pentadecane C₁₅H₃₂ alkane, which may be used as jet fuel, at a range of approximately 1% in the dry cell weight of flocculated cells.

Conclusion

This is the first study on biofuel production using flocculated cells in which the specific manner of production may be regulated by cultivation conditions.

     

Interactions between climatic and production factors on returns of female pigs to service during summer in Japanese commercial breeding herds

The objectives of this study were to determine the associations between climatic factors and production factors for returns to service of female pigs during summer and to quantify the associations between these factors and occurrences of returns to service. The factors that were assessed were as follows: maximum temperature (HT), relative humidity, age of gilts at first mating, parity, weaning-to-first-mating interval (WMI), and lactation length. The study analyzed records of 18,307 gilts in 99 herds and 78,135 parity records of 56,322 sows in 103 herds; all the females were first-serviced between June and September, 2007 to 2009. Average daily HT and relative humidity for 15 days post-service of a female were obtained from 21 local weather stations and coordinated with the performance data of the respective local herds. The returns to service were categorized into three groups: regular returns (RRs: 18–24 days), irregular returns (IRs: 25–38 days), and late returns to service (LRs: 39 days or later). Two-level mixed-effects models were applied to the data by using a herd at level 2 and an individual record at level 1. In mated gilts, the occurrences (%) of RRs, IRs, and LRs were 4.8%, 1.8%, and 5.3%, respectively. In mated sows, the respective occurrences were 3.3%, 1.8%, and 4.2%. Mean values (ranges) of HT and relative humidity were 28.4 °C (13.6 °C–39.8 °C) and 73.4% (35.0%–98.0%), respectively. In gilts, as HT increased from 25 °C to 30 °C, the occurrence of RR increased from 3.7% to 4.4% (P < 0.05). However, there was no association between the occurrence of RR and either relative humidity (P = 0.17) or age at first mating (P = 0.23). In addition, there were no associations between the occurrences of either gilt IR or LR and HT (P ≥ 0.05), relative humidity (P ≥ 0.46), or age at first mating (P ≥ 0.32). In sows, greater occurrences of RRs, IRs, and LRs were associated with higher HT, lower parity, and a WMI of 7 days or longer (P < 0.05), but they were not associated with relative humidity (P ≥ 0.62) or lactation length (P ≥ 0.13). The occurrence of RRs in sows of all WMI groups increased 1.22 (1.04⁵) times for each 5 °C increase in HT. For sows with WMI 0 to 6 days, each 5 °C rise in HT increased the occurrence of IRs and LRs by 1.36 (1.06⁵) and 1.27 (1.05⁵) times, respectively. However, there was no association between increased HT and occurrences of IRs or LRs for sows with WMI 7 days or longer (P ≥ 0.38). Therefore, in order to prevent returns to service, it is recommended that producers apply cooling management for females during the post-service periods in summer.     

Polysaccharide-polynucleotide complexes (15): thermal stability of schizophyllan (SPG)/poly(C) triple strands is controllable by alpha-amino acid modification

Schizophyllan (SPG), a beta-1,3-glucan polysaccharide which is known to form macromolecular complexes with certain polynucleotides, was modified by a reductive amination method with alpha-amino acids (Arg, Lys, and Ser). The thermal stability of the complexes as estimated by T(m) was enhanced in SPG-Arg and SPG-Lys conjugates which have pI values higher than the pH of the medium (8.0). The T(m) shift increased with the increase in the percentage of alpha-amino acid introduced and the highest T(m) values attained were 64 degrees C for SPG-Arg conjugate and 62 degrees C for SPG-Lys conjugate, which are higher by 13 and 11 degrees C, respectively, than those of the unmodified SPG+poly(C) complex. In the SPG-Ser conjugate with a pI lower than the medium pH (8.0), the T(m) values decreased with an increase in the percentage of Ser. Formation of the macromolecular complex was no longer detected above 13.2% Ser. The findings indicate that the T(m) values are easily controllable by the type and percentage of the introduced alpha-amino acids. We believe, therefore, that the present conjugates, consisting of naturally originated SPG and alpha-amino acids, provide an important lead for developing nontoxic artificial vectors and to control the affinity with polynucleotides in response to medium pH and temperature.     

Production of embryonic and fetal-like red blood cells from human induced pluripotent stem cells

We have previously shown that human embryonic stem cells can be differentiated into embryonic and fetal type of red blood cells that sequentially express three types of hemoglobins recapitulating early human erythropoiesis. We report here that we have produced iPS from three somatic cell types: adult skin fibroblasts as well as embryonic and fetal mesenchymal stem cells. We show that regardless of the age of the donor cells, the iPS produced are fully reprogrammed into a pluripotent state that is undistinguishable from that of hESCs by low and high-throughput expression and detailed analysis of globin expression patterns by HPLC. This suggests that reprogramming with the four original Yamanaka pluripotency factors leads to complete erasure of all functionally important epigenetic marks associated with erythroid differentiation regardless of the age or the tissue type of the donor cells, at least as detected in these assays. The ability to produce large number of erythroid cells with embryonic and fetal-like characteristics is likely to have many translational applications.     

ClC-3 chloride channels facilitate endosomal acidification and chloride accumulation

We investigated the involvement of ClC-3 chloride channels in endosomal acidification by measurement of endosomal pH and chloride concentration [Cl-] in control versus ClC-3-deficient hepatocytes and in control versus ClC-3-transfected Chinese hamster ovary cells. Endosomes were labeled with pH or [Cl-]-sensing fluorescent transferrin (Tf), which targets to early/recycling endosomes, or alpha2-macroglobulin (alpha2M), which targets to late endosomes. In pulse label-chase experiments, [Cl-] was 19 mM just after internalization in alpha2M-labeled endosomes in primary cultures of hepatocytes from wild-type mice, increasing to 58 mM over 45 min, whereas pH decreased from 7.1 to 5.4. Endosomal acidification and [Cl-] accumulation were significantly impaired in hepatocytes from ClC-3 knock-out mice, with [Cl-] increasing from 16 to 43 mM and pH decreasing from 7.1 to 6.0. Acidification and Cl- accumulation were blocked by bafilomycin. In Tf-labeled endosomes, [Cl-] was 46 mM in wild-type versus 35 mM in ClC-3-deficient hepatocytes at 15 min after internalization, with corresponding pH of 6.1 versus 6.5. Approximately 4-fold increased Cl- conductance was found in alpha2M-labeled endosomes isolated from hepatocytes of wild-type versus ClC-3 null mice. In contrast, Golgi acidification was not impaired in ClC-3-deficient hepatocytes. In transfected Chinese hamster ovary cells expressing ClC-3A, endosomal acidification and [Cl-] accumulation were enhanced. [Cl-] in alpha2M-labeled endosomes was 42 mM (control) versus 53 mM (ClC-3A) at 45 min, with corresponding pH 5.8 versus 5.2; [Cl-] in Tf-labeled endosomes at 15 min was 37 mM (control) versus 49 mM (ClC-3A) with pH 6.3 versus 5.9. Our results provide direct evidence for involvement of ClC-3 in endosomal acidification by Cl- shunting of the interior-positive membrane potential created by the vacuolar H+ pump.     

Simple method of designing a bilobed flap

In the present study, we devised a new method of designing bilobed flaps. This method makes the flap easy to draw and has the merits of diffusing the tension on the flap and minimizing the dog-ear. We used this flap in 16 patients with face and head skin defects and obtained good results.