Rapid and flexible scale-down media development and optimization for perfusion culture

Perfusion culture is steadily gaining interest due to advantages such as higher cell densities, increased flexibility, higher volumetric output, decreased product retention times, and more consistent product. Perfusion culture systems are drastically different than fed-batch cultures as they allow bioreactors to continuously run for prolonged periods of time by constantly feeding fresh media while removing spent media. This drastic process difference means that typical fed-batch medium is not optimal for perfusion culture. Instead, perfusion culture media is most optimal when it fits within culture process constraints such as vessel volume exchanges per day. Irvine Scientific has rapidly developed custom media for a specific perfusion culture process that resulted in a significant increase in viable cell density and antibody titer. The custom media was developed in the following steps: (1) identification of initial base media, (2) development and verification of a scale-down perfusion model, (3) optimization of media components, and (4) parsing down extraneous components while maintaining growth and titer. First, a media survey was conducted in batch cultures to determine an initial basal media composition to further refine. From a panel of over 20 different media, top candidate media were selected based on improved viable cell density, viability, titer, and glycan profile over the control medium. Next, a scale-down perfusion model was developed and validated using the control medium. Mini bioreactor tubes were used as culture vessels and daily perfusion was conducted by centrifugation, aspiration, and replenishment with fresh medium. Antibody glycan profiles from the scale-down perfusion model matched profiles from large-scale bioreactor perfusion cultures, thus validating the use of this scale-down model. Although this system is not controlled, the pH was monitored offline and stayed within range. The candidate media were evaluated in the scale-down perfusion model and the top prospects in terms of titer and viable cell density were chosen for further development by changing the composition of amino acids, metals, and other components. Lastly, amino acid concentrations were further optimized while parsing down extraneous components according to spent media analysis from the previous experiment. The top medium resulted in a significant increase in viable cell density and titer compared to the original control medium. This process created a custom medium for perfusion culture that significantly increased viable cell density and antibody titer while maintaining a desired glycan profile. The media development process in scale-down perfusion cultures was rapid and took about 10 weeks. Development in controlled, scale-down bioreactor systems may be more costly and take longer as procedures, protocols, and methods first need to be transferred and verified. This scale-down perfusion model, while manual, is an easy and flexible way to develop perfusion medium with efficient throughput. Currently, the medium is being verified in perfusion bioreactors and can be further improved. In the future, we will investigate increasing the productivity by concentrating the culture medium while decreasing the volume vessel exchanges

Chromospheric Activity of HAT-P-11: an Unusually Active Planet-Hosting K Star

Kepler photometry of the hot Neptune host star HAT-P-11 suggests that its spot latitude distribution is comparable to the Sun's near solar maximum. We search for evidence of an activity cycle in the CaII H & K chromospheric emission $S$-index with archival Keck/HIRES spectra and observations from the echelle spectrograph on the ARC 3.5 m Telescope at APO. The chromospheric emission of HAT-P-11 is consistent with a $\gtrsim 10$ year activity cycle, which plateaued near maximum during the Kepler mission. In the cycle that we observed, the star seemed to spend more time near active maximum than minimum. We compare the $\log R^\prime_{HK}$ normalized chromospheric emission index of HAT-P-11 with other stars. HAT-P-11 has unusually strong chromospheric emission compared to planet-hosting stars of similar effective temperature and rotation period, perhaps due to tides raised by its planet.Comment: 16 pages, 8 figures; accepted to the Astrophysical Journa

Aksesibiliti Terhadap Fasiliti Bagi Penjawat Awam Dalam Golongan Orang Kelainan Upaya (OKU) Di Bangunan Kerajaan, Kuching

Kajian ini dijalankan untuk meneroka pengalaman tiga pegawai OKU yang berkhidmat dengan kerajaan negeri Sarawak. Seramai tiga orang pegawai berlainan jenis kecacatan dan berkhidmat di tiga bangunan kerajaan yang berlainan telah di temubual secara semi-berstruktur dan data telah dianalisis menggunakan Analisis Bertema. Hasil dapatan menunjukkan tema utama adalah berkaitan dengan persepsi terhadap kemudahan sedia ada, penerimaan majikan dan rakan sekerja serta cabaran terhadap kemudahan dan fasiliti di tempat kerja. Implikasi serta cadangan terhadap penambahbaikan aksesibiliti fasiliti untuk golongan kurang upaya di bangunan kerajaan turut dibincangkan

Chromosome-Level Genome Assembly of Chinese Sucker (Myxocyprinus asiaticus) Reveals Strongly Conserved Synteny Following a Catostomid-Specific Whole-Genome Duplication

publishedVersio

Idiopathic ileal volvulus with multiple concomitant infections in a starving man

Background: Small bowel volvulus is defined as the torsion of the small intestine, potentially leading to bowel obstruction, gangrene and perforation. It is a rare condition, especially in adults. Case presentation: A 30-year-old man was retrieved from the jungle with severe weight loss and abdominal symptoms. He succumbed to death despite 22 days of intensive medical treatment. An autopsy revealed a ruptured gangrenous ileal volvulus with peritonitis and subdiaphragmatic abscess. Further laboratory analysis detected systemic Candida tropicalis and intestinal gramnegative bacterial sepsis, systemic Zika virus viremia, leptospirosis complicating rhabdomyolysis and disseminated intravascular coagulopathy, Type I Herpes Simplex virus infection of the tongue and upper gastrointestinal tract. The cause of death was the ruptured ileal volvulus, complicated with upper gastrointestinal bleeding due to Herpes simplex virus esophagitis in a malnourished patient with resolving leptospirosis and underlying Zika virus co-infection. Conclusion: Rare clinical scenarios of adult-onset intestinal volvulus with concomitant multiple infections precludes clinical diagnosis and early treatment, leading to devastating consequences of clinical outcome. The positive clinical and postmortem correlation is a good learning lesson in many disciplines of medicine and science

Wars2 is a determinant of angiogenesis.

Coronary flow (CF) measured ex vivo is largely determined by capillary density that reflects angiogenic vessel formation in the heart in vivo. Here we exploit this relationship and show that CF in the rat is influenced by a locus on rat chromosome 2 that is also associated with cardiac capillary density. Mitochondrial tryptophanyl-tRNA synthetase (Wars2), encoding an L53F protein variant within the ATP-binding motif, is prioritized as the candidate at the locus by integrating genomic data sets. WARS2(L53F) has low enzyme activity and inhibition of WARS2 in endothelial cells reduces angiogenesis. In the zebrafish, inhibition of wars2 results in trunk vessel deficiencies, disordered endocardial-myocardial contact and impaired heart function. Inhibition of Wars2 in the rat causes cardiac angiogenesis defects and diminished cardiac capillary density. Our data demonstrate a pro-angiogenic function for Wars2 both within and outside the heart that may have translational relevance given the association of WARS2 with common human diseases

Roles of Small GTPase Rac1 in the Regulation of Actin Cytoskeleton during Dengue Virus Infection

An important clinical characteristic of dengue hemorrhagic fever/dengue shock syndrome is increased vascular permeability. Actin cytoskeleton is a significant element of endothelial barrier function regulation. In vitro study showed that dengue virus infection could induce redistributions of actin cytoskeleton. It is not precisely clear the roles of actin and the mechanisms of its reorganization during the infection. Using immunochemical assays, drug inhibition assays and protein interaction profiling methods, we aimed to identify the ways in which dengue virus serotype 2 interacts with actin cytoskeleton. The study showed that dynamic treadmilling of actin is necessary for dengue virus entry, production and release, while small GTPase Rac1 also plays multiple roles during these processes. In addition, we demonstrated the association of viral E protein with actin, indicating a direct effect of viral protein on the structural modifications of actin cytoskeleton. Our results provide evidence for the participation of Rac1 signaling pathways in viral protein-induced actin reorganizations, which may be a mechanism involved in the etiology of dengue hemorrhagic fever

Affectus Hispaniae en la historiografía del Alto Imperio

This paper analyses texts written by Greek and Latin High Empire historians dealing with Hispania. Some of the authors have a very positive view (Florus, Iustinus, Appian) while others are clearly negative (Veleius Paterculus, Valerius Maximus) though most of them show little interest, indifference or variety of opinions. When there is interest in the region or praise, it is because the author comes from Hispania or he is trying to please an emperor born in Hispania, but it could also be due to a universal conception of history revealing a critical attitude towards Roman imperialism, as in Appian. The praise found in Iustinus’s epitome should be attributed to the author of the epitome rather than to Pompeius Trogus. This can be taken as evidence for situating Iustinus’s life and work in the 2nd century A.D. Loathing of Hispania seems to have its origins in conservative, ‘optimate’ nationalist circles, who perceive the province as the ‘popular’ region that acclaimed and welcomed ‘seditious’ individuals such as Tiberius Gracchus and Sertorius.Se estudian en este trabajo los textos de historiadores del Alto Imperio, latinos y griegos, que tratan sobre Hispania. En algunos autores encontramos una visión muy positiva (Floro, Justino, Apiano) y en otros claramente negativa (Veleyo Patérculo, Valerio Máximo), aunque en la mayoría de los casos hay escasa atención, indiferencia o diversidad de opiniones. El interés por la región y los elogios pueden estar motivados por el origen hispánico del autor o su voluntad de agradar a algún emperador oriundo de Hispania, pero también por una concepción universal de la historia que denota en ocasiones una posición crítica con el imperialismo romano, como es el caso de Apiano. La alabanza que hallamos en el epítome de Justino creemos que debe atribuirse más al epitomador que a Pompeyo Trogo, lo que apoyaría una datación temprana de la vida y la obra de Justino (s. II d.C.). La aversión hacia Hispania parece haber surgido en medios conservadores, “optimates” nacionalistas, que ven la provincia como el territorio “popular”, que encumbró y acogió a “sediciosos” como Tiberio Graco y Sertorio

TESS spots a mini-neptune interior to a hot saturn in the TOI-2000 system

Hot jupiters (P 60 $\mathrm{M}_\oplus$) are almost always found alone around their stars, but four out of hundreds known have inner companion planets. These rare companions allow us to constrain the hot jupiter's formation history by ruling out high-eccentricity tidal migration. Less is known about inner companions to hot Saturn-mass planets. We report here the discovery of the TOI-2000 system, which features a hot Saturn-mass planet with a smaller inner companion. The mini-neptune TOI-2000 b ($2.70 \pm 0.15 \,\mathrm{R}_\oplus$, $11.0 \pm 2.4 \,\mathrm{M}_\oplus$) is in a 3.10-day orbit, and the hot saturn TOI-2000 c ($8.14^{+0.31}_{-0.30} \,\mathrm{R}_\oplus$, $81.7^{+4.7}_{-4.6} \,\mathrm{M}_\oplus$) is in a 9.13-day orbit. Both planets transit their host star TOI-2000 (TIC 371188886, V = 10.98, TESS magnitude = 10.36), a metal-rich ([Fe/H] = $0.439^{+0.041}_{-0.043}$) G dwarf 174 pc away. TESS observed the two planets in sectors 9-11 and 36-38, and we followed up with ground-based photometry, spectroscopy, and speckle imaging. Radial velocities from CHIRON, FEROS, and HARPS allowed us to confirm both planets by direct mass measurement. In addition, we demonstrate constraining planetary and stellar parameters with MIST stellar evolutionary tracks through Hamiltonian Monte Carlo under the PyMC framework, achieving higher sampling efficiency and shorter run time compared to traditional Markov chain Monte Carlo. Having the brightest host star in the V band among similar systems, TOI-2000 b and c are superb candidates for atmospheric characterization by the JWST, which can potentially distinguish whether they formed together or TOI-2000 c swept along material during migration to form TOI-2000 b.Comment: v3 adds RV frequency analysis; 25 pages, 11 figures, 14 tables; revision submitted to MNRAS; machine-readable tables available as ancillary files; posterior samples available from Zenodo at https://doi.org/10.5281/zenodo.7683293 and source code at https://doi.org/10.5281/zenodo.798826