Improving product quality and productivity of bispecific molecules through the application of continuous perfusion principles.

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.

Perfusion CHO cell culture applied to lower aggregation and increase volumetric productivity for a bispecific recombinant protein.

Secreted recombinant proteins can aggregate during cell culture. We studied a poorly-behaved bispecific scaffold that increasingly aggregated (up to 62% high molecular weight species, HMW) as a function of culture time in a fed-batch and intensified cell culture processes. We identified that protein aggregates increased with accumulated protein concentration inside the bioreactor.

Failure to Thrive: An Expanded Differential Diagnosis.

The patient is a term 6-month-old male, who presented with failure to thrive since birth. History was remarkable for suspected milk and soy protein allergy, gastroesophageal reflux, constipation, and abdominal distension that was present since birth. He was losing weight despite oral intake of over 100 kcal/kg per day.

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology.

Generating a highly productive cell line is resource intensive and typically involves long timelines because of the need to screen large numbers of candidates in protein production studies. This has led to miniaturization and automation strategies to allow for reductions in resources and higher throughput. Current approaches rely on the use of standard cell culture vessels and bulky liquid handling equipment.

Analysis of Tubespins as a suitable scale-down model of bioreactors for high cell density CHO cell culture.

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.

Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease.

Background: Bapineuzumab, a humanized anti-amyloid-beta monoclonal antibody, is in clinical development for the treatment of Alzheimer's disease.

Methods: We conducted two double-blind, randomized, placebo-controlled, phase 3 trials involving patients with mild-to-moderate Alzheimer's disease--one involving 1121 carriers of the apolipoprotein E (APOE) ε4 allele and the other involving 1331 noncarriers. Bapineuzumab or placebo, with doses varying by study, was administered by intravenous infusion every 13 weeks for 78 weeks.

Monoclonal antibody disulfide reduction during manufacturing: Untangling process effects from product effects.

Manufacturing-induced disulfide reduction has recently been reported for monoclonal human immunoglobulin gamma (IgG) antibodies, a widely used modality in the biopharmaceutical industry. This effect has been tied to components of the intracellular thioredoxin reduction system that are released upon cell breakage. Here, we describe the effect of process parameters and intrinsic molecule properties on the extent of reduction.

A voxel-based analysis of FDG-PET in traumatic brain injury: regional metabolism and relationship between the thalamus and cortical areas.

The objective was to study the correlations and the differences in glucose metabolism between the thalamus and cortical structures in a sample of severe traumatic brain injury (TBI) patients with different neurological outcomes. We studied 49 patients who had suffered a severe TBI and 10 healthy control subjects using 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET). The patients were divided into three groups: a vegetative or minimally-conscious state (MCS&VS) group (n=17), which included patients who were in a vegetative or a minimally conscious state; an In-post-traumatic amnesia (In-PTA) group (n=12), which included patients in PTA; and an Out-PTA group (n=20), which included patients who had recovered from PTA.

Voxel-based statistical analysis of thalamic glucose metabolism in traumatic brain injury: relationship with consciousness and cognition.

Objective: To study the relationship between thalamic glucose metabolism and neurological outcome after severe traumatic brain injury (TBI).

Methods: Forty-nine patients with severe and closed TBI and 10 healthy control subjects with (18)F-FDG PET were studied. Patients were divided into three groups: MCS&VS group (n = 17), patients in a vegetative or a minimally conscious state; In-PTA group (n = 12), patients in a state of post-traumatic amnesia (PTA); and Out-PTA group (n = 20), patients who had emerged from PTA.

[Thalamic metabolism and neurological outcome after traumatic brain injury. A voxel-based morphometric FDG-PET study].

Objective: to study the relationship between thalamic metabolism and neurological outcome in patients who had sustained a traumatic brain injury (TBI).

Methods: nineteen patients who had sustained a severe TBI and ten control subjects were included in this study. Six of the 19 patients had a low level of consciousness (vegetative state or minimally conscious state), while thirteen showed normal consciousness.

A randomized, placebo-controlled study to assess the efficacy and safety of 3 doses of paliperidone palmitate in adults with acutely exacerbated schizophrenia.

This study assessed the efficacy and the safety of a dosing regimen that was revised from earlier studies for the investigational injectable atypical antipsychotic paliperidone palmitate (approved in the USA, August 2009) for adult patients with acutely exacerbated schizophrenia. The patients (N = 652) were randomly assigned (1:1:1:1) to paliperidone palmitate at 25, 100, or 150 mg eq. or placebo in this 13-week double-blind study.

Efficacy and safety of paliperidone palmitate in adult patients with acutely symptomatic schizophrenia: a randomized, double-blind, placebo-controlled, dose-response study.

This 13-week, double-blind study evaluated the efficacy and safety of the atypical antipsychotic paliperidone palmitate (recently approved in the United States) versus placebo administered as monthly gluteal injections (after two initial doses given 1 week apart) in acutely symptomatic patients with schizophrenia. Patients (N=388) were randomly assigned (1 : 1 : 1 : 1) to paliperidone palmitate 50, 100, or 150 mg eq. or placebo.

Increased amygdala and parahippocampal gyrus activation in schizophrenic patients with auditory hallucinations: an fMRI study using independent component analysis.

Objective: Hallucinations in patients with schizophrenia have strong emotional connotations. Functional neuroimaging techniques have been widely used to study brain activity in patients with schizophrenia with hallucinations or emotional impairments. However, few of these studies have investigated the association between hallucinations and emotional dysfunctions using an emotional auditory paradigm.

Multicomponent MR Image Denoising.

Magnetic Resonance images are normally corrupted by random noise from the measurement process complicating the automatic feature extraction and analysis of clinical data. It is because of this reason that denoising methods have been traditionally applied to improve MR image quality. Many of these methods use the information of a single image without taking into consideration the intrinsic multicomponent nature of MR images.

Large scale refolding and purification of the catalytic domain of human BACE-2 produced in E. coli.

Evidence for a key role of beta-amyloid (Abeta) in Alzheimer's disease has led to considerable interest in potential therapeutic strategies targeting enzymes involved in processing the amyloid precursor protein (APP). Beta-site APP Cleaving Enzyme (BACE or beta-secretase) is a membrane bound aspartyl protease that has been shown to be directly involved in Abeta production and, therefore, is at the forefront of therapeutic targets in the treatment of Alzheimer's disease. BACE-2, an enzyme closely related to BACE, regulates Abeta production in a manner antagonistic to BACE, suggesting that non-selective inhibition of BACE-2 by BACE inhibitors might impair the lowering of Abeta.

MR and genetics in schizophrenia: focus on auditory hallucinations.

Although many structural and functional abnormalities have been related to schizophrenia, until now, no single biological marker has been of diagnostic clinical utility. One way to obtain more valid findings is to focus on the symptoms instead of the syndrome. Auditory hallucinations (AHs) are one of the most frequent and reliable symptoms of psychosis.

Robust MRI brain tissue parameter estimation by multistage outlier rejection.

This article addresses the problem of the tissue type parameter estimation in brain MRI in the presence of partial volume effects. Automatic MRI brain tissue classification is hampered by partial volume effects that are caused by the finite resolution of the acquisition process. Due to this effect intensity distributions in brain MRI cannot be well modeled by a simple mixture of Gaussians and therefore more complex models have been developed.

MRI denoising using non-local means.

Magnetic Resonance (MR) images are affected by random noise which limits the accuracy of any quantitative measurements from the data. In the present work, a recently proposed filter for random noise removal is analyzed and adapted to reduce this noise in MR magnitude images. This parametric filter, named Non-Local Means (NLM), is highly dependent on the setting of its parameters.

High yield expression of human BACE constructs in Eschericia coli for refolding, purification, and high resolution diffracting crystal forms.

BACE (beta-site APP cleaving enzyme) or beta-secretase, the enzyme responsible for processing APP to give the N-terminal portion of the Abeta peptide, is a membrane bound aspartyl protease consisting of an ectodomain catalytic unit, a C-terminal transmembrane segment and a cytoplasmic domain. Three BACE constructs, pET11a-BACE, pQE80L-BACE, and pQE70-BACE were designed to terminate at a position just before the transmembrane domain (Ser(432)) and are described schematically below. (1) pET11a-T7.

Schizophrenia with auditory hallucinations: a voxel-based morphometry study.

Many studies have shown widespread but subtle pathological changes in gray matter in patients with schizophrenia. Some of these studies have related specific alterations to the genesis of auditory hallucinations, particularly in the left superior temporal gyrus, but none has analysed the relationship between morphometric data and a specific scale for auditory hallucinations. The present study aims to define the presence and characteristics of structural abnormalities in relation with the intensity and phenomenology of auditory hallucinations by means of magnetic resonance voxel-based morphometry (MR-VBM) method applied on a highly homogeneous group of 18 persistent hallucinatory patients meeting DSM-IV criteria for schizophrenia compared to 19 healthy matched controls.

Chronic auditory hallucinations in schizophrenic patients: MR analysis of the coincidence between functional and morphologic abnormalities.

Purpose: To prospectively evaluate if functional magnetic resonance (MR) imaging abnormalities associated with auditory emotional stimuli coexist with focal brain reductions in schizophrenic patients with chronic auditory hallucinations.

Materials And Methods: Institutional review board approval was obtained and all participants gave written informed consent. Twenty-one right-handed male patients with schizophrenia and persistent hallucinations (started to hear hallucinations at a mean age of 23 years +/- 10, with 15 years +/- 8 of mean illness duration) and 10 healthy paired participants (same ethnic group [white], age, and education level [secondary school]) were studied.

A nonparametric MRI inhomogeneity correction method.

Magnetic resonance images are commonly affected by intensity inhomogeneities which make it difficult to obtain any quantitative measures from them. We present a new method of automatically correcting this artifact using a nonparametric coarse to fine approach which allows bias fields to be modeled with different frequency ranges without user supervision. We also propose a new entropy-related cost function based on the combination of intensity and gradient image features for more robust homogeneity measurement.

Emotional words induce enhanced brain activity in schizophrenic patients with auditory hallucinations.

Neuroimaging studies of emotional response in schizophrenia have mainly used visual (faces) paradigms and shown globally reduced brain activity. None of these studies have used an auditory paradigm. Our principal aim is to evaluate the emotional response of patients with schizophrenia to neutral and emotional words.

Emotional auditory paradigm in neuroimaging: a base for the study of psychosis.

Introduction: Since the arrival of neuroimaging numerous studies have tried to analyze the differences between emotional and non-emotional response. The majority of these studies use visual approach (faces) and begin with data in normal subjects. The present study introduces a new paradigm for the study of emotional response based on auditory approach and designed specifically for the study of psychoses.