Algorithm for Improved Stereoscopic Cloud-Top Height Retrieval Based on Visible and Infrared Bands for Himawari-8 and FY-4A

Stereoscopic cloud-top height (CTH) retrieval from two geostationary (GEO) satellites is usually realized through a visible (VIS) band with a high horizontal resolution. A stereoscopic-based CTH retrieval algorithm (prototype dual-GEO CTH algorithm) proposed in our previous study also adopts this approach. Although this approach can retrieve accurate stereoscopic CTHs, the heights of optically thin upper clouds overlying the lower clouds are challenging to retrieve because the parallax difference between two GEOs is determined by the lower clouds owing to the low reflectance from the upper clouds. To address this problem, this paper proposes an improved stereoscopic CTH retrieval algorithm, named the improved dual-GEO CTH algorithm, for Himawari-8 and FengYun (FY)-4A GEOs. The proposed algorithm employs an infrared (IR) band in addition to a VIS band. A seamless image cloning technique is adopted to blend the VIS and IR images, which are then used to retrieve the stereoscopic CTHs. The retrieved CTHs are compared with the Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) and Cloud Profiling Radar (CPR) CTHs for three occasions involving upper clouds overlying lower clouds. Results show that the proposed algorithm outperforms the prototype dual-GEO CTH algorithm in the case of upper clouds overlying lower clouds. Notably, although the proposed algorithm is intended for Himawari-8 and FY-4A GEOs, it can be easily extended to any combination of two GEOs.

Cloning of Bacillus subtilis phytase gene construct in Escherichia coli

Background and Objectives: Phytase has a hydrolysis function of phytic acid, which yields inorganic phosphate. Bacillus species can produce thermostable alkaline phytase. The aim of this study was to isolate and clone a Phytase gene (Phy) from Bacillus subtilis in Escherichia coli. Materials and Methods: In this study, the extracellular PhyC gene was isolated from Bacillus subtilis Phytase C. After purification of the bands, DNA fragment of Phy gene was cloned by T/A cloning technique, and the clone was transformed into Escherichia coli. Afterward, the pGEM-Phy was transferred into E. coli Top-10 strain and the recombinants were plated on LB agar containing 100 µg/ml ampicillin. The colonization of 1171 bp of gene Phytase C was confirmed by PCR. The presence of gene-targeting in vector was confirmed with enzymatic digestion by XhoI and XbaI restriction enzymes. Results: The Phytase gene was successfully cloned in E. coli. The result of cloning of 1171 bp Phytase gene was confirmed by PCR assay. Conclusion: Our impression of this article is that several methods, such as using along with microbial, plant phytase reproduction, or low-phytic acid corn may be the better way from a single phytase.

Simplified plasmid cloning with a universal MCS design and bacterial in vivo assembly

Background The ability to clone DNA sequences quickly and precisely into plasmids is essential for molecular biology studies. The recent development of seamless cloning technologies has made significant improvements in plasmid construction, but simple and reliable tools are always desirable for time- and labor-saving purposes. Results We developed and standardized a plasmid cloning protocol based on a universal MCS (Multiple Cloning Site) design and bacterial in vivo assembly. With this method, the vector is linearized first by PCR (Polymerase Chain Reaction) or restriction digestion. Then a small amount (10 ~ 20 ng) of this linear vector can be mixed with a PCR-amplified insert (5× molar ratio against vector) and transformed directly into competent E. coli cells to obtain the desired clones through in vivo assembly. Since we used a 36-bp universal MCS as the homologous linker, any PCR-amplified insert with ~ 15 bp compatible termini can be cloned into the vector with high fidelity and efficiency. Thus, the need for redesigning insert-amplifying primers according to various vector sequences and the following PCR procedures was eliminated. Conclusions Our protocol significantly reduced hands-on time for preparing transformation reactions, had excellent reliability, and was confirmed to be a rapid and versatile plasmid cloning technique. The protocol contains mostly mixing steps, making it an extremely automation-friendly and promising tool in modern biology studies.

THE GENERAL COMPLEXITY OF THE PROBLEM TO RECOGNIZE HAMILTONIAN PATHS

Generic-case approach to algorithmic problems has been offered by A. Miasnikov, V. Kapovich, P. Schupp, and V. Shpilrain in 2003. This approach studies an algorithm behavior on typical (almost all) inputs and ignores the rest of inputs. In this paper, we study the generic complexity of the problem of recognition of Hamiltonian paths in finite graphs. A path in graph is called Hamiltonian if it passes through all vertices exactly once. We prove that under the conditions P 6= NP and P = BPP for this problem there is no polynomial strongly generic algorithm. A strongly generic algorithm solves a problem not on the whole set of inputs, but on a subset, the sequence of frequencies of which exponentially quickly converges to 1 with increasing size. To prove the theorem, we use the method of generic amplification, which allows to construct generically hard problems from the problems hard in the classical sense. The main component of this method is the cloning technique, which combines the inputs of a problem together into sufficiently large sets of equivalent inputs. Equivalence is understood in the sense that the problem is solved similarly for them.

TRACING THE KINSHIP OF BRUCELLA SP. BY USINGA POLYMERASE CHAIN REACTION TECHNIQUE WITH A SHORT PRIMER

Brucellosis is a zoonotic disease that has spread throughout the world and has an important impacton both human and animal health. The four species of Brucella that cause disease in humans are Brucellaabortus, B. suis, B. melitensis and B. canis, and B. melitensis as the most pathogenic species. This Researchused 46 samples were collected from Government Small Ruminants Abattoir in Bogor Regency. Thirty twospleen samples were examined by previous research and showed a positive result when were tested withCFT and PCR techniques, but sequencing has not yet been done. Fourteen serum and spleen samples wereexamined by the similar techniques. The Research aimed to determined the genetic relationship of Brucellasp. using a PCR technique with a specific short primer to B. mellitensis. Cloning technique was appliedpreviously to five PCR positive samples before sequencing. Cloning and sequencing result of the Sample91 showed higher homology to B. melitensis and B. abortus for 127 nucleotide lengths, 97.6% -100% and99.2% -100% respectively. In the phylogenic tree, the Sample 91 was part of B melitensis sequences 1, 2,and 3 with accession numbers LT962930.1 and LT962936.1, B abortus sequences 1 and 2 with accessionnumbers CP033079.1 and B. abortus sequence 1 with accession number CP034695.1. Sample of 95, 97, 7,and 13 have lower homologies than Sample 91.

Pathological features of cloned calves that died in the neonatal period

ABSTRACT: Somatic-cell nuclear transfer is a cloning technique that enables the creation of a viable embryo from a donor adult to produce a genetically identical individual. This technique opens numerous potential possibilities for medicine and animal reproduction. However, several reports have documented cloning-related issues. Embryo and fetal losses remain significantly higher than in other techniques, and there is a high incidence of dystocia and hydrops, which decreases efficiency and increases costs. Animals delivered at term often exhibit a syndrome known as macrosomia and experience difficulties in adapting to life outside the uterus, and death is a common outcome. In the present study, 41 cloned calves that died in the neonatal period were subjected to gross and histopathological examination. Most important gross lesions were found in the liver (enlargement, congestion, yellowish color), kidneys (brownish color at surface and cut, and cysts), lungs (atelectasis, parenchymal consolidation, and secretions in bronchi and bronchioles), and heart (concentric and eccentric hypertrophy, hematic cysts, persistence of ductus arteriosus). Primary microscopic findings were seen in the liver, kidneys, and lungs from neonatal calves. In the liver, 85% of the animals exhibited hepatic degeneration. The presence of a brownish pigment within the cortical tubules of the kidneys was found in approximately 90% of the samples; the presence of this pigment has not been previously reported in cloned calves. In the lungs, a large number of animals exhibiting lesions characteristic of pneumonia (55%). These changes were the pivotal causes of death, mainly due to problems in adapting to life outside the uterus and opportunistic infections in the neonatal period. Further investigation focusing on pathological anatomical changes is necessary to map these abnormalities in cloned animals.

An Integrative Synthetic Biology Approach to Interrogating Cellular Ubiquitin and Ufm Signaling

Global identification of substrates for PTMs (post-translational modifications) represents a critical but yet dauntingly challenging task in understanding biology and disease pathology. Here we presented a synthetic biology approach, namely ‘YESS’, which coupled Y2H (yeast two hybrid) interactome screening with PTMs reactions reconstituted in bacteria for substrates identification and validation, followed by the functional validation in mammalian cells. Specifically, the sequence-independent Gateway® cloning technique was adopted to afford simultaneous transfer of multiple hit ORFs (open reading frames) between the YESS sub-systems. In proof-of-evidence applications of YESS, novel substrates were identified for UBE3A and UFL1, the E3 ligases for ubiquitination and ufmylation, respectively. Therefore, the YESS approach could serve as a potentially powerful tool to study cellular signaling mediated by different PTMs.