Generating and characterizing a comprehensive panel of CHO cells glycosylation mutants for advancing glycobiology and biotechnology research.

This report describes the development and characterization of a comprehensive collection of CHO cell glycosylation mutants with significant potential for advancing glycobiology and biotechnology. EPO-Fc and trastuzumab, two model molecules, were produced using these mutants to assess the effects of mutated glycogenes, and LC-MS/MS analysis was employed to quantitatively analyse their N-glycans. EPO-Fc exhibited exclusively homogeneous Man9 glycans only when nearly all α-mannosidases in the genome were inactivated, except lysosomal MAN2B1.

Development of a Machine Learning Algorithm to Correlate Lumbar Disc Height on X-rays with Disc Bulging or Herniation.

Lumbar disc bulging or herniation (LDBH) is one of the major causes of spinal stenosis and related nerve compression, and its severity is the major determinant for spine surgery. MRI of the spine is the most important diagnostic tool for evaluating the need for surgical intervention in patients with LDBH. However, MRI utilization is limited by its low accessibility.

Improving Antibody Production in Stably Transfected CHO Cells by CRISPR-Cas9-Mediated Inactivation of Genes Identified in a Large-Scale Screen with Chinese Hamster-Specific siRNAs.

The Chinese hamster ovary (CHO) cell line is commonly used for the production of biotherapeutics. As cell productivity directly affects the cost of production, methods are developed to manipulate the expression of specific genes that are known to be involved in protein synthesis, folding, and secretion to increase productivity. However, there are no large-scale CHO-specific functional screens to identify novel gene targets that impact the production of secreted recombinant proteins.

Attenuated glutamine synthetase as a selection marker in CHO cells to efficiently isolate highly productive stable cells for the production of antibodies and other biologics.

Chinese hamster ovary (CHO) cells are the biopharmaceutical industry's primary means of manufacturing therapeutic proteins, including monoclonal antibodies. The major challenge in cell line development for the production of recombinant biopharmaceuticals lies in generating and isolating rare high-producing stable clones, amongst thousands of low-producing or unstable clones, in a short period of time. One approach to accomplish this is to use the glutamine synthetase (GS) selection system, together with the GS inhibitor, methionine sulfoximine (MSX).

Acute Infarction in the Artery of Percheron Distribution during Cerebral Angiography: A Case Report and Literature Review.

Improvements in techniques, contrast agents, and catheter design have significantly decreased angiography-related neurological deficits and complications. This article reports a case involving an angiographic total obliteration arteriovenous malformation (AVM) in a patient with an acute infarction in the artery of Percheron (AOP) distribution following angiography. Furthermore, imaging of an AOP acute infarction in cerebral angiography is presented.

The "less-is-more" in therapeutic antibodies: Afucosylated anti-cancer antibodies with enhanced antibody-dependent cellular cytotoxicity.

Therapeutic monoclonal antibodies are the fastest growing class of biological therapeutics for the treatment of various cancers and inflammatory disorders. In cancer immunotherapy, some IgG1 antibodies rely on the Fc-mediated immune effector function, antibody-dependent cellular cytotoxicity (ADCC), as the major mode of action to deplete tumor cells. It is well-known that this effector function is modulated by the N-linked glycosylation in the Fc region of the antibody.

Comprehensive MDCT evaluation of patients with suspected May-Thurner syndrome.

Objective: The purpose of this essay is to introduce the MDCT protocol and interpretation techniques for optimal evaluation of patients with suspected May-Thurner syndrome.

Conclusion: May-Thurner syndrome is always the working diagnosis when a patient presents with unilateral left lower limb swelling without signs of infection. MDCT is useful for fast, comprehensive evaluation of the vascular system to determine whether May-Thurner syndrome or an alternative condition is present.

Collapsin response mediator proteins (CRMPs) are a new class of microtubule-associated protein (MAP) that selectively interacts with assembled microtubules via a taxol-sensitive binding interaction.

Collapsin response mediator proteins are ubiquitously expressed from multiple genes (CRMPs 1-5) and play important roles in dividing cells and during semaphorin 3A (Sema3A) signaling. Nonetheless, their mode of action remains opaque. Here we carried out in vivo and in vitro assays that demonstrate that CRMPs are a new class of microtubule-associated protein (MAP).

Comprehensive evaluation of patients suspected with deep vein thrombosis using indirect CT venography with multi-detector row technology: from protocol to interpretation.

With the increased availability of multi-detector row CT, indirect CT venography has become an important non-invasive image modality for patients with suspected deep vein thrombosis. Use of indirect CT venography can not only diagnose/exclude deep vein thrombosis, but can also determine if there are other anomalies or diseases which might contribute to the patient's symptoms. In this pictorial essay, we introduce the scanning protocol, post-processing techniques, and interpretation algorithm used in widely available 64 multi-detector row technology.

Arteriovenous fistula and graft evaluation in hemodialysis patients using MDCT: a primer.

Objective: Patent arteriovenous fistula (AVF) is related to better prognosis and quality of life for patients on long-term dialysis. When AVF dysfunction is suspected, MDCT is a good noninvasive tool for evaluating the entire AVF structure and determining reversible conditions for treatment. The aim of this article is to introduce the scanning and interpretation techniques and to illustrate the conditions related to early and late fistula failures.

Comprehensive evaluation of CT pulmonary angiography for patients suspected of having pulmonary embolism.

Recently, CT pulmonary angiography (CTPA), especially performed with multi-detector row CT, has become a key imaging modality for pulmonary embolism. However, CTPA that was performed under clinical suspicion of pulmonary embolism has been shown to lead to high prevalence of alternative diagnosis, up to 25.4%.

Cytoplasmic ACK1 interaction with multiple receptor tyrosine kinases is mediated by Grb2: an analysis of ACK1 effects on Axl signaling.

ACK1 (activated Cdc42-associated kinase 1), a cytoplsmic tyrosine kinase, is implicated in metastatic behavior, cell spreading and migration, and epidermal growth factor receptor (EGFR) signaling. The function of ACK1 in the regulation of receptor tyrosine kinases requires a C-terminal region that demonstrates a significant homology to the EGFR binding domain of MIG6. In this study, we have identified additional receptor tyrosine kinases, including Axl, leukocyte tyrosine kinase, and anaplastic lymphoma kinase, that can bind to the ACK1/MIG6 homology region.

Comprehensive evaluation of patients with suspected renal hypertension using MDCT: from protocol to interpretation.

Objective: The objectives of this article are to, first, describe the reasons for and details of the MDCT protocol for patients with suspected renal hypertension; second, explain the importance of comprehensive evaluation by MDCT in patients with suspected renal hypertension; third, review the image appearances of important conditions that may be encountered in the reader's clinical practice; and, fourth, explain what information should be included in a comprehensive MDCT report for patients with suspected renal hypertension.

Conclusion: MDCT is widely used for renal artery evaluation in patients with resistant hypertension. Because the regions outside the renal arteries might also have diseases that contribute to the symptoms, a comprehensive interpretation including the renal arteries, renal parenchyma, adrenal glands, and scanned abdomen is very important.

MDCT evaluation of congenital mitral-aortic intervalvular fibrosa aneurysm: implications for the aetiology and differential diagnosis.

Mitral-aortic intervalvular fibrosa aneurysm is a rare disease whose aetiology remains a matter of debate. Here we present the youngest reported patient with the disease, a 6-month-old boy, without a history of infection, which supports a congenital origin as initially proposed. Multidetector-row CT (MDCT) surpassed echocardiography in delineating the intracardiac anatomical details with high spatial resolution, confirming the important problem-solving role of MDCT in the diagnosis of congenital heart disease.

Neonatal cardiac multidetector row CT: why and how we do it.

Neonatal congenital heart disease is a most difficult area of diagnostic radiology because of the small patient body size and fast resting heart rate. Recently, the spatial and temporal resolution of multidetector-row CT (MDCT) has evolved so that neonatal congenital heart disease can be precisely diagnosed. We describe the role of MDCT in neonatal congenital heart disease and offer tips for the scanning procedure to familiarize radiologists with this developing field.

Vertebral and carotid artery anomalies in patients with aberrant right subclavian arteries.

Background: There is little published evidence regarding the patterns and prevalence of vertebral artery (VA) and common carotid artery (CCA) anomalies in patients with an aberrant right subclavian artery (ARSCA).

Objective: To study the patterns and prevalence of VA and CCA anomalies in patients with ARSCA.

Materials And Methods: In a 2-year period we reviewed the children referred with suspected vascular ring who had undergone multidetector-row CT.

Homogeneous enhancement in pediatric thoracic CT aortography using a novel and reproducible method: contrast-covering time.

Objective: This study compares the empiric setting and contrast-covering time (CCT) concept for i.v. contrast injection in pediatric thoracic CT aortography.

Kinectin-dependent assembly of translation elongation factor-1 complex on endoplasmic reticulum regulates protein synthesis.

Kinectin is an integral membrane protein with many isoforms primarily found on the endoplasmic reticulum. It has been found to bind kinesin, Rho GTPase, and translation elongation factor-1delta. None of the existing models for the quaternary organization of the elongation factor-1 complex in higher eukaryotes involves kinectin.

An engineered microenvironment for multidimensional microscopy of live cells.

Multidimensional imaging (MD) of live cells is gaining importance in biomedical research as the commercial availability of confocal, nonlinear optical microscopes, environmental chambers, and specific fluorescence probes grows. One crucial aspect of the MD live cell imaging involves the proper immobilization of cells, which refers to the rapid and sufficient immobilization of cells on the microscope stage, neither disrupting the cellular structure and functions nor affecting the optical properties of the cells and the environments. Conventional cell immobilization methods glue the anchoring cells to coated surfaces, but such methods require centrifugation or extended incubation and are not suitable for cells in suspension.

Engineering microenvironment for expansion of sensitive anchorage-dependent mammalian cells.

Tissue engineering involves ex vivo seeding of anchorage-dependent mammalian cells onto scaffolds, or transplanting cells in vivo. The cell expansion currently requires repeated cell detachment from solid substrata by enzymatic, chemical or mechanical means. The report here presents a high yield three-dimensional culture and harvest system circumventing the conventional detachment requirements.

TGF-beta1 regulation in hepatocyte-NIH3T3 co-culture is important for the enhanced hepatocyte function in 3D microenvironment.

Co-culture of hepatocytes or hepatocyte spheroids with the supporting NIH3T3 in a 3D microcapsule formed with a hybrid natural/synthetic matrix has led to enhanced hepatocyte functions. We investigated the mechanism of the functional enhancement in co-culture with respect to the contributions of soluble factors and direct cell-cell interactions. The conditioned media from the co-culture induced higher P450 cytochrome oxidase activity (indicated by EROD assay) in the microencapsulated hepatocytes than the conditioned media from the NIH3T3- or the hepatocytes-alone controls.