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Treatment for acute kidney injury (AKI) is suboptimal. A better understanding of the pathogenesis of AKI may lead to new therapeutic approaches. Kidney transcriptomics of folic acid‐induced AKI (FA‐AKI) in mice identified Runx1 as the most upregulated RUNX family gene. We then examined the expression of RUNX1 in FA‐AKI, in bacterial lipopolysaccharide (LPS)‐induced cytokine storm‐AKI (CS‐AKI), and in human AKI. In cultured mouse tubule cells, we explored the expression and role of RUNX1 in response to the cytokine TWEAK or LPS. A chemical inhibitor of RUNX1 (Ro5‐3335) was used in animal models of AKI to test its potential as a therapeutic target. RUNX1 overexpression in FA‐AKI was validated at the mRNA and protein levels and localized mainly to tubule cell nuclei. CS‐AKI also upregulated kidney RUNX1. Increased tubule and interstitial RUNX1 expression were also observed in human AKI. In cultured mouse tubule cells, the pro‐inflammatory cytokine TWEAK and LPS increased RUNX1 and IL‐6 expression. Mechanistically, RUNX1 bound to the Il6 gene promoter and RUNX1 targeting with the chemical inhibitor Ro5‐3335, or a specific small interfering RNA (siRNA), prevented the TWEAK‐ and LPS‐induced upregulation of IL6 through a RUNX1/NFκB1 p50 pathway. In vivo, preventive Ro5‐3335 improved kidney function and reduced inflammation in FA‐AKI and CS‐AKI. However, Ro5‐3335 administration after the insult only improved kidney function in CS‐AKI. Kidney transcriptomics identified inflammatory genes and transcription factor mRNAs such as Yap1 and Trp53 as key targets of Ro5‐3335 in CS‐AKI. In conclusion, RUNX1 contributes to AKI by driving the expression of genes involved in inflammation and represents a novel therapeutic target in AKI. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland. [ABSTRACT FROM AUTHOR]

We report results from 8 h of JWST /MIRI low resolution spectroscopic (LRS) monitoring directly followed by 7 h of JWST /NIRSpec prism spectroscopic monitoring of the benchmark binary brown dwarf WISE 1049AB, the closest, brightest brown dwarfs known. We find water, methane, and CO absorption features in both components, including the 3.3  μ m methane absorption feature and a tentative detection of small grain (⁠|$\lt $| 1 μ m) silicate absorption at |$\gt $| 8.5 μ m in WISE 1049A. Both components vary significantly (⁠|$\gt 1~{{\rm per\ cent}}$|⁠), with WISE 1049B displaying larger variations than WISE 1049A. Using K -means clustering, we find three main transition points in wavelength for both components of the binary: (1) change in behaviour at |$\sim$| 2.3 μ m coincident with a CO absorption bandhead, (2) change in behaviour at 4.2 μ m, close to the CO fundamental band at |$\lambda \gt $|  4.4 µm, and (3) change in behaviour at 8.3–8.5 µm, potentially corresponding to silicate absorption. We interpret the light curves observed with both NIRSpec and MIRI as likely stemming from (1) a deep pressure level driving the double-peaked variability seen in WISE 1049B at wavelengths |$\lt $| 2.3 and |$\gt $| 8.5 µm, (2) an intermediate pressure level shaping the light-curve morphology between 2.3 and 4.2 µm, and (3) a higher altitude pressure level producing single-peaked and plateaued light-curve behaviour between 4.2 and 8.5 µm. [ABSTRACT FROM AUTHOR]

Planar cell polarity (PCP) proteins coordinate tissue morphogenesis by governing cell patterning and polarity. Asymmetrically localized on the plasma membrane of cells, transmembrane PCP proteins are trafficked by endocytosis, suggesting they may have intracellular functions that are dependent or independent of their extracellular role, but whether these functions extend to transcriptional control remains unknown. Here, we show the nuclear localization of transmembrane, PCP protein, VANGL2, in the HCC1569 breast cancer cell line, and in undifferentiated, but not differentiated, HC11 cells that serve as a model for mammary lactogenic differentiation. The loss of Vangl2 function results in upregulation of pathways related to STAT5 signaling. We identify DNA binding sites and a nuclear localization signal in VANGL2, and use CUT&RUN to demonstrate recruitment of VANGL2 to specific DNA binding motifs, including one in the Stat5a promoter. Knockdown (KD) of Vangl2 in HC11 cells and primary mammary organoids results in upregulation of Stat5a, Ccnd1 and Csn2, larger acini and organoids, and precocious differentiation; phenotypes are rescued by overexpression of Vangl2, but not Vangl2ΔNLS. Together, these results advance a paradigm whereby PCP proteins coordinate tissue morphogenesis by keeping transcriptional programs governing differentiation in check. [ABSTRACT FROM AUTHOR]

Fatigue and musculoskeletal pain are also frequent in patients with psoriasis (PsO) without arthritis (PsA). The current study aimed to assess the impact of an intervention program based on aerobic training to reduce fatigue and musculoskeletal pain in patients with PsO without PsA. A total of 118 male patients with PsO volunteered in the current interventional study and were randomly allocated to the experimental (n = 59) or control group (n = 59). The intervention consisted of a 16-week aerobic training program on a treadmill, three sessions per week, consisting of a warm-up, 35–50 min treadmill exercise (increasing 5 min/4 weeks) at a work intensity of 50–65% of peak heart-rate (increasing 5%/4 weeks), and cooling-down. The functional assessment of chronic illness therapy fatigue scale (FACIT-Fatigue), health assessment questionnaire disability index (HAQ-DI), and visual analog scale (VAS) were compared pre and post intervention. Nutritional intake, maximal aerobic power, lipid profile, serum markers of muscle damage, and body composition were also assessed. When compared to baseline, FACIT-Fatigue, HAQ-DI, and VAS scores were significantly improved without increasing markers of muscle damage. Fat mass percentage, lipid profile, and maximal oxygen consumption were also improved. In conclusion, a 16-week aerobic training program at moderate intensity was safe, well tolerated, and effective in psoriatic patients without PsA. Long-term follow-up studies are required to examine whether these promising results may improve clinical outcomes. [ABSTRACT FROM AUTHOR]

Purpose: To analyze the clinical findings, management, and results of oncological treatments and to identify prognostic factors in patients diagnosed with head and neck sarcoma. Methods: We performed a retrospective analysis including 48 adult patients with primary head and neck sarcomas, treated between 2006 and 2018 in a referral center of Argentina. Results: The median follow-up time was 44 months (range: 4–146 months). Five-year overall survival was 68%. On univariate analysis, radiation-induced sarcomas (p=0.038) had worse survival. Five-year disease-free survival was 58% and local recurrence at 2 years was 22.7%. On multivariate analysis, positive/close resection margins (p=0.031), radiation-induced sarcomas (p=0.037), and mandibular and oral cavity location (p=0.002) were independent prognostic factors associated to local recurrence and shorter disease-free survival. Conclusion: Head and neck sarcomas are a rare entity. Surgery is feasible in more than 80% of patients, with an acceptable overall and disease-free survival. Radiation-induced sarcomas, location in the mandible and oral cavity, and close margins were significant prognostic factors in our population. [ABSTRACT FROM AUTHOR]

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Cell line development (CLD) represents a critical, yet time‐consuming, step in the biomanufacturing process as significant resources are devoted to the scale‐up and screening of several hundreds to thousands of single‐cell clones. Typically, transfected pools are fully recovered from selection and characterized for growth, productivity, and product quality to identify the best pools suitable for single‐cell cloning (SCC) using limiting dilution or fluorescence‐activated cell sorting (FACS). Here we report the application of the Berkeley Lights Beacon Instrument (BLI) in an early SCC process to accelerate the CLD timeline. Transfected pools were single‐cell cloned when viabilities reached greater than 85% or during selection when viabilities were less than 30%. Clones isolated from these accelerated processes exhibited comparable growth, productivity, and product quality to those derived from a standard CLD process and fit into an existing manufacturing platform. With these approaches, up to a 30% reduction in the overall CLD timeline was achieved. Furthermore, early process‐derived clones demonstrated equivalent long‐term stability compared with standard process‐derived clones over 50 population doubling levels (PDLs). Taken together, the data supported early SCC on the BLI as an attractive approach to reducing the standard CLD timeline while still identifying clones with acceptable manufacturability. [ABSTRACT FROM AUTHOR]

Background: Disorders of the temporomandibular joint (TMJ) are frequent and are usually associated with other disorders of the facial skeleton. Surgery might be needed to correct TMJ anatomy and function and, in cases where pathologies coexist, a two-stage corrective surgery might be needed. However, the current fashion of single-stage procedures is feasible with the aid of new technologies such as computer-assisted surgery (CAS). This is a step forward toward performing complex procedures such as a TMJ replacement with simultaneous orthognathic surgery. CAS allows designing patient-fitted prosthesis and more predictable and accurate surgeries. Moreover, intraoperative development can be controlled in real time with intraoperative navigation, and postoperative results can be measured and compared afterwards. Aims: The primary purpose of this article is to present the protocol used in our institution for orthognathic surgery associated with unilateral and bilateral TMJ replacement with patient-fitted prostheses guided with CAS. Materials and methods: We present two cases to illustrate our protocol and its results. Results: In the first case, the difference in millimeters between planning and surgical outcomes was 1.72 mm for the glenoid component and 2.16 mm for the condylar prosthesis; for the second case, differences in the right side were 2.59 mm for the glenoid component and 2.06 mm for the ramus, and in the left side, due to the anatomy the difference was a little greater, without clinical significance Conclusion: Combined surgery of the midface and mandible with total TMJ replacement is feasible and beneficial for the patient. CAS facilitates the planning and design of custom-fit prosthesis and execution of these procedures. [ABSTRACT FROM AUTHOR]

Bispecific protein scaffolds can be more complex than traditional monoclonal antibodies (MAbs) because two different sites/domains for epitope binding are needed. Because of this increased molecular complexity, bispecific molecules are difficult to express and can be more prone to physical and chemical degradation compared to MAbs, leading to higher levels of protein aggregates, clipped species, or modified residues in cell culture. In this study, we investigated cell culture performance for the production of three types of bispecific molecules developed at Amgen. In particular, we cultured a total of six CHO cell lines in both an approximately 12‐day fed‐batch process and an approximately 40‐day high‐density perfusion process. Harvested cell culture fluid from each process was purified and analyzed for product quality attributes including aggregate levels, clipped species, charge variants, individual amino acid modifications and host cell protein (HCP) content. Our studies showed that in average, the intensified perfusion process increased 15‐fold the integrated viable cell density and the total harvested product (and fivefold the daily volumetric productivity) compared to fed‐batch. Furthermore, bispecific product quality improved in perfusion culture (as analyzed in affinity‐capture pools) with reduction in levels of aggregates (up to 72% decrease), clipped species (up to 75% decrease), acidic variants (up to 76% decrease), deamidated/isomerized species in complementarity‐determining regions, and HCP (up to 84% decrease). In summary, the intensified perfusion process exhibited better productivity and product quality, highlighting the potential to use it as part of a continuous manufacturing process for bispecific scaffolds. [ABSTRACT FROM AUTHOR]

Therapeutic bispecific antibodies are formed by assembly of multichain polypeptides. In general, a bispecific antibody has two different light chains and two different heavy chains that fold and correctly pair via engineered interchain interactions. Because of some incorrect assembly, product‐related impurities can be prevalent (e.g., half molecules, mispaired light chains, homodimers), requiring its removal during subsequent purification. In this study, we investigated the modulation of impurity levels in a stable Chinese hamster ovary cell line X expressing a bispecific antibody A formed by two light chains (LC1 and LC2) and two heavy chains (HC1 and HC2) that assembled intracellularly into a heterodimer (LC1–HC1 + LC2–HC2) via engineered charged residues. Cell line X exhibited the best volumetric productivity, growth, and viability in culture compared with other clones but also showed higher levels of half antibody species (>10%); therefore, to minimize process yield loss, better understanding, and control of impurity formation was pursued. We found this cell line decreased half antibody levels from 16% to 1% when temperature changed from 36°C to 32.5°C or 31.5°C. However, lower temperature also increased high‐molecular‐weight (HMW) species from 4% to 12%. To determine the impurity species composition, we characterized enriched fractions with half antibody or HMW. Intact mass spectrometry analysis revealed half antibody was LC2–HC2, whereas HMW was a mixture with ~50% as LC1–HC1 homodimer. Results suggested LC2–HC2 was easily folded and could be secreted as half antibody, especially at 36°C. On the contrary, LC1–HC1 was more susceptible to misfold or aggregate, a phenomenon more acute for cell line X at lower culture temperature because of 60% increased LC1 and HC1 messenger RNA levels. Although temperature modulation was cell line X‐specific, the propensity of LC2–HC2 to form half antibodies and LC1–HC1 to aggregate appeared in other cell lines also expressing bispecific antibody A, suggesting an amino‐acid sequence‐dependent mechanism. In summary, impurity formation in cell line X was temperature‐dependent and was influenced by different molecule characteristics between the LC1–HC1 and LC2–HC2 parts. Ultimately, we selected a biphasic cell culture process with a growth phase followed by a lower temperature phase to improve product quality and purification yield. Gomez and coworkers studied the formation of product‐related impurities in a CHO cell line expressing a bispecific antibody formed by two light chains (LC1, LC2) and two heavy chains (HC1, HC2). In particular, cell culture temperature modulated formation of half bispecific antibody (half B‐Ab) and high molecular weight (HMW) species. More uniquely, both impurity levels were inversely correlated with increasing 1 to 16% half B‐Ab and decreasing 12 to 4% HMW with increasing temperature. [ABSTRACT FROM AUTHOR]

The Renewable Energies for Galápagos (ERGAL) project was launched in Ecuador as part of the “Zero fossil fuels on the Galápagos Islands” initiative. The ERGAL project strategy includes the “Jatropha for Galápagos” pilot project, which seeks to generate electricity from jatropha oil (JO) extracted from wild jatropha bushes, which are harvested by small-scale jatropha suppliers. Using life-cycle analysis, this study assessed the environmental impact of electricity produced from this biofuel under three systems: a blend system (BS) with 20% JO and 80% diesel, a reference system (RS) consisting of 100% diesel and a jatropha system (JS) made up of JO. The results show that the BS had slightly better environmental performance than RS, but worse than JS. Under the BS, the main environmental loads resulted from diesel combustion in electricity generation and the transportation of the raw material. An advantage of the BS is the low impact resulting from jatropha production, given that fertilizers are not used. However, this production strategy is highly dependent upon the presently limited yields from jatropha bushes. To increase the environmental sustainability of the BS, the logistics of transporting the fruits and improving their yields are key issues that should be considered. Finally, the low environmental impact of JS is due to diesel not being used in electricity production. However, fuel use for transportation of the high volumes of raw material and the refined product, coupled with the use of the purification substance as part of the refining are major contributors to environmental loads of JS. [ABSTRACT FROM AUTHOR]

Lysophosphatidic acid (LPA) affects several female reproductive functions through G-protein-coupled receptors. LPA contributes to embryo implantation via the lysophospholipid LPA3 receptor. In the present study we investigated the participation of endogenous LPA signalling through the LPA3 receptor in vascularisation and decidualisation, two crucial events at the maternal-fetal interface. Pregnant rats were treated with diacylglycerol pyrophosphate (DGPP), a highly selective antagonist of LPA3 receptors, on Day 5 of gestation. Pregnant rats received intrauterine (i.u.) injections of single doses of DGPP (0.1 mg kg-1) in a total volume of 2 µL in the left horn (treated horn) in the morning of GD5. DGPP treatment produced aberrant embryo spacing and increased embryo resorption. The LPA3 receptor antagonist decreased the cross-sectional length of the uterine and arcuate arteries and induced histological anomalies in the decidua and placentas. Marked haemorrhagic processes, infiltration of immune cells and tissue disorganisation were observed in decidual and placental tissues from sites of resorption. The mRNA expression of three vascularisation markers, namely interleukin 10 (Il10), vascular endothelial growth factor (Vegfa) and vascular endothelial growth factor receptor 1 (Vegfr1), was reduced at sites of resorption from Day 8. The results show that the disruption of endogenous LPA signalling by blocking the LPA3 receptor modified the development of uterine vessels with consequences in the formation of the decidua and placenta and in the growth of embryos. [ABSTRACT FROM AUTHOR]

High cell density (HCD) culture increases recombinant protein productivity via higher biomass. Compared to traditional fed-batch cultures, HCD is achieved by increased nutrient availability and removal of undesired metabolic components via regular medium replenishment. HCD process development is usually performed in instrumented lab-scale bioreactors (BR) that require time and labor for setup and operation. To potentially minimize resources and cost during HCD experiments, we evaluated a 2-week 50-mL Tubespin (TS) simulated HCD process where daily medium exchanges mimic the medium replacement rate in BR. To best assess performance differences, we cultured 13 different CHO cell lines in simulated HCD as satellites from simultaneous BR, and compared growth, metabolism, productivity and product quality. Overall, viability, cell-specific productivity and metabolism in TS were comparable to BR, but TS cell growth and final titer were lower by 25 and 15% in average, respectively. Peak viable cell densities were lower in TS than BR as a potential consequence of lower pH, different medium exchange strategy and dissolved oxygen limitations. Product quality attributes highly dependent on intrinsic molecule or cell line characteristics (e.g., galactosylation, afucosylation, aggregation) were comparable in both scales. However, product quality attributes that can change extracellularly as a function of incubation time (e.g., deamidation, C-terminal lysine, fragmentation) were in general lower in TS because of shorter residence time than HCD BR. Our characterization results and two case studies show that TS-simulated HCD cultures can be effectively used as a simple scale-down model for relative comparisons among cell lines for growth or productivity (e.g., clone screening), and for investigating effects on protein galactosylation. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:490-499, 2017 [ABSTRACT FROM AUTHOR]

Sarcoidosis is a systemic disease of unknown etiology characterized by the presence of noncaseating granulomas in any organ, most commonly the lungs and intrathoracic lymph nodes. A diagnosis of sarcoidosis should be suspected in any young or middle-aged adult presenting with unexplained cough, shortness of breath, or constitutional symptoms, especially among blacks or Scandinavians. Diagnosis relies on three criteria: (1) a compatible clinical and radiologic presentation, (2) pathologic evidence of noncaseating granulomas, and (3) exclusion of other diseases with similar findings, such as infections or malignancy. An early and accurate diagnosis of sarcoidosis remains challenging, because initial presentations may vary, many patients are asymptomatic, and there is no single reliable diagnostic test. Prognosis is variable and depends on epidemiologic factors, mode of onset, initial clinical course, and specific organ involvement. The optimal treatment for sarcoidosis remains unclear, but corticosteroid therapy has been the mainstay of therapy for those with significantly symptomatic or progressive pulmonary disease or serious extrapulmonary disease. Refractory or complex cases may require immunosuppressive therapy. Despite aggressive treatment, some patients may develop life-threatening pulmonary, cardiac, or neurologic complications from severe, progressive disease. End-stage disease may ultimately require lung or heart transplantation for eligible patients. [ABSTRACT FROM AUTHOR]

Video streaming services have been developed using different approaches. Peer-to-peer (P2P) streaming is a competitive technique in the distribution of video contents, thanks to its collaborative and sharing features. Nowadays, several applications distribute television contents over a P2P platform, broadcasting plenty of channels to an increasing audience. The performance of such applications over wireless networks is still an open issue, considering the highly variable environment, and the scarce resources. Most of these applications are developed to work properly over wired networks. This paper presents a performance evaluation model of the traffic behavior through an analytical representation, resembling the network interactions during a video transmission. The model, useful in determining the throughput parameter, does accurately evaluate the network performance. The analytical model is validated through the emulation mode in NS-3 simulator. [ABSTRACT FROM PUBLISHER]

Anti-mitotic therapies have been considered a hallmark in strategies against abnormally proliferating cells. Focusing on the extensively studied family of thiosemicarbazone (TSC) compounds, we have previously identified 4,4’-dimethoxybenzophenone thiosemicarbazone (T44Bf) as a promising pharmacological compound in a panel of human leukemia cell lines (HL60, U937, KG1a and Jurkat). Present findings indicate that T44Bf-mediated antiproliferative effects are associated with a reversible chronic mitotic arrest caused by defects in chromosome alignment, followed by induced programmed cell death. Furthermore, T44Bf selectively induces apoptosis in leukemia cell lines when compared to normal peripheral blood mononuclear cells. The underlying mechanism of action involves the activation of the mitochondria signaling pathway, with loss of mitochondrial membrane potential and sustained phosphorylation of anti-apoptotic protein Bcl-xL as well as increased Bcl-2 (enhanced phosphorylated fraction) and pro-apoptotic protein Bad levels. In addition, ERK signaling pathway activation was found to be a requisite for T44Bf apoptotic activity. Our findings further describe a novel activity for a benzophenone thiosemicarbazone and propose T44Bf as a promising anti-mitotic prototype to develop chemotherapeutic agents to treat acute leukemia malignancies. [ABSTRACT FROM AUTHOR]

Cyclic AMP represents one of the most studied signaling molecules and its role in proliferation and differentiation processes has been well established. Intracellular cAMP levels are tightly regulated where the MRP4 transporter plays a major role. In the present study, we sought to establish whether cAMP modulated MRP4 expression in pancreatic adenocarcinoma cell lines. Quantitative PCR and western blot studies showed that cAMP-increasing agents enhanced MRP4 transcripts and protein levels in PANC-1 cells. Reporter luciferase experiments carried out in pancreatic AR42J cells showed that intracellular cAMP up-regulates MRP4 through an Epac2- and Rap1- mediated mechanism whereas extracellular cAMP reduced MRP4 promoter activity by a MEK/ERK-mediated pathway. Present results show that cAMP regulates MRP4 promoter activity, and further indicate that the balance between intracellular and extracellular cAMP levels determines MRP4 expression. [ABSTRACT FROM AUTHOR]

Copper concentration can impact lactate metabolism in Chinese Hamster ovary (CHO) cells. In our previous study, a 20-fold increase in initial copper concentration enabled CHO cultures to shift from net lactate production to net lactate consumption, and achieve higher cell growth and productivity. In this follow-up study, we used transcriptomics to investigate the mechanism of action (MOA) of copper that mediates this beneficial metabolism shift. From microarray profiling (days 0-7), the number of differentially expressed genes increased considerably after the lactate shift (>day 3). To uncouple the effects of copper at early time points (days 0-3) from that of lactate per se (>day 3), and to validate microarray hits, we analyzed samples before the lactate shift by RNA-Seq. Out of 6,398 overlapping genes analyzed by both transcriptomic methods, only the early growth response 1 gene-coding for a transcription factor that activates signaling pathways in response to environmental stimuli-satisfied the differential expression criteria (fold change ≥1.5; P < 0.05). Gene expression correlation and biological pathway analyses further confirmed that copper differences exerted minimal transcriptional impact on the CHO cultures before the lactate shift. By contrast, genes associated with hypoxia network and oxidative stress response were upregulated after the lactate shift. These upregulations should boost cell proliferation and survival, but do not account for the preceding shift in lactate metabolism. The findings here indicate that the primary MOA of copper that enabled the shift in lactate metabolism is not at the transcriptional level. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:429-442, 2014 [ABSTRACT FROM AUTHOR]

During production of therapeutic monoclonal antibodies (mAb), it is highly desirable to remove and control antibody aggregates in the manufacturing process to minimize the potential risk of immunogenicity to patients. During process development for the production of a recombinant IgG in a CHO cell line, we observed atypical high variability from 1 to 20% mAb aggregates formed during cell culture that negatively impacted antibody purification. Analytical characterization revealed the IgG aggregates were mediated by hydrophobic interactions likely caused by misfolded antibody during intracellular processing. Strikingly, data analysis showed an inverse correlation of lower cell culture temperature producing higher aggregate levels. All cultures at 37°C exhibited ≤5% aggregates at harvest. Aggregate levels increased 4-12-fold in 33°C cultures when compared to 37°C, with a corresponding 2-4-fold increase in heavy chain (HC) and light chain (LC) mRNA. Additionally, 37°C cases showed a greater excess of LC to HC mRNA levels. Endoplasmic reticulum (ER) chaperone expression and ER size also increased 25-75% at 33°C versus 37°C but to a lesser extent than LC and HC mRNA, consistent with a potential limiting ER folding capacity at 33°C for this cell line. Finally, we identified a 2-5-fold increase in mAb aggregate formation at 33°C compared to 37°C cultures for three additional CHO cell lines. Taken together, our observations indicate that low culture temperature can increase antibody aggregate formation in CHO cells by increasing LC and HC transcripts coupled with limited ER machinery. Biotechnol. Bioeng. 2012;109: 125-136. © 2011 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

THIOMABs are recombinant antibodies with reactive cysteine residues used for forming THIOMAB-drug conjugates (TDCs). We recently reported a new impurity associated with THIOMABs: one of the engineered cysteines forms a disulfide bond with an extra light chain (LC) to generate a triple light chain antibody (3LC). In our previous investigations, increased LC expression increased 3LC levels, whereas increased glutathione (GSH) production decreased 3LC levels. In this work, on three stably transfected CHO cell lines, we investigated the effects of temperature, pH, dissolved oxygen (DO), and hydrolysate on 3LC formation during THIOMAB fed-batch cell culture production. Although pH between 6.8 and 7.0 had no significant impact on 3LC formation, temperature at 35°C instead of 33 or 31°C generated the lowest 3LC values for two cell lines. The decreased 3LC level correlated with increased GSH production. We implemented a 35°C temperature process for large-scale (2,000 L) production of a THIOMAB. This process reduced 3LC levels by ∼50% compared with a 33°C temperature process. By contrast, DO and hydrolysate had modest effect on 3LC levels for the model cell line studied. Overall, we did not find significant changes in LC expression under the conditions tested, whereas changes in GSH production were more evident. By investigating the impact of bioreactor process and medium conditions on 3LC levels, we identified strategies that reduced 3LC levels. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [ABSTRACT FROM AUTHOR]

The article presents a case study of a 64-year-old man who encountered thromboembolic events (TE) associated with antitumor necrosis factor-alpha treatment.

Investigates how anthropogenic edges may alter bird-mediated seed dispersal from edge to forest interior as a function of edge age and presence of treefall gaps in a montane tropical forest in southwestern Colombia. Fruit abundance; Capture rates of frugivorous birds; Potential seed movement; Implications for conservation.

Presents a study which looked at the influence of anthropogenic edges on the distribution of understory birds in a neotropical montane forest. Detailed information on the methodology used to conduct the study; Discussion based on the results of the study.

Dysregulation of histone deacetylases (HDAC) has been proposed as a potential contributor to aberrant transcriptional profiles that can lead to changes in cognitive functions. It is known that METH negatively impacts the prefrontal cortex (PFC) leading to cognitive decline and addiction whereas modafinil enhances cognition and has a low abuse liability. We investigated if modafinil (90 mg/kg) and methamphetmine (METH) (1 mg/kg) may differentially influence the acetylation status of histones 3 and 4 (H3ac and H4ac) at proximal promoters of class I, II, III, and IV HDACs. We found that METH produced broader acetylation effects in comparison with modafinil in the medial PFC. For single dose, METH affected H4ac by increasing its acetylation at class I Hdac1 and class IIb Hdac10, decreasing it at class IIa Hdac4 and Hdac5. Modafinil increased H3ac and decreased H4ac of Hdac7. For mRNA, single-dose METH increased Hdac4 and modafinil increased Hdac7 expression. For repeated treatments (4 d after daily injections over 7 d), we found specific effects only for METH. We found that METH increased H4ac in class IIa Hdac4 and Hdac5 and decreased H3/H4ac at class I Hdac1, Hdac2, and Hdac8. At the mRNA level, repeated METH increased Hdac4 and decreased Hdac2. Class III and IV HDACs were only responsive to repeated treatments, where METH affected the H3/H4ac status of Sirt2, Sirt3, Sirt7, and Hdac11. Our results suggest that HDAC targets linked to the effects of modafinil and METH may be related to the cognitive-enhancing vs cognitive-impairing effects of these psychostimulants. [ABSTRACT FROM AUTHOR]