Critical quality attributes (CQAs) of a therapeutic antibody produced from integrated continuous bioprocessing

The integrated continuous bioprocess provides an innovative way to produce protein drugs with flexibility and efficiency. However, during the long-term cultivation and complicated production, how to ensure the process stability and product quality is critically important. In this study, the monoclonal antibody (mAb) was produced in a bioreactor operated in a perfusion mode utilizing the ATF cell retention system for up to 32 days. The 2L harvest per day starting at day 10 was continuously purified using the 3-column periodic counter-current (PCC) chromatography system. The first protein A capture purification was performed with the dynamic binding capacity of 50% breakthrough around 60 mg mAb/mL of resin (vs 20 mg/mL resin for batch purification) for 120 cycles or 360 column operations followed by a polishing step of mixed mode chromatography for 20 cycles. The process and quality attributes were monitored daily. The results demonstrate consistency in both the purification process and the mAb qualities (in the aspects of product integrity, aggregates, and glycan profile) between PCC and batch purifications. Culture-related charge heterogeneity was observed accompanied by an increase of bioreactor harvest time using both batch and PCC purification processes. In addition, the impurities such as endotoxin and HCP were also monitored while under this high capacity utilization of chromatography resins. By sharing the insights of process and quality attributes, we hope to provide better understanding on the process-related heterogeneity between batch and continuous production and/or purification

Differences of Various Region-of-Interest Methods for Measuring Dopamine Transporter Availability Using T

This study was to investigate whether various region-of-interest (ROI) methods for measuring dopamine transporter (DAT) availabilities by single photon emission computed tomography (SPECT) are statistically different, whether results of medical research are thereby influenced, and causes of these differences. Eighty-four healthy adults with Tc99m-TRODAT-1 SPECT and magnetic resonance imaging (MRI) scans were included. Six major analysis approaches were compared: (1) ROI drawn on the coregistered MRI; (2) ROIs drawn on the SPECT images; (3) standard ROI templates; (4) threshold-ROIs; (5) atlas-based mappings with coregistered MRI; and (6) atlas-based mappings with SPECT images. Using the atlas-based approaches we assessed the influence of striatum ROIs by slice-wise and voxel-wise comparisons. In (5) and (6), three partial-volume correction (PVC) methods were also explored. The results showed that DAT availabilities obtained from different methods were closely related but quite different and leaded to significant differences in determining the declines of DAT availability per decade (range: 5.95–11.99%). Use of 3D whole-striatum or more transverse slices could avoid biases in measuring the striatal DAT declines per decade. Atlas-based methods with PVC may be the preferable methods for medical research

Application and comparison of scoring indices to predict outcomes in patients with healthcare-associated pneumonia

Introduction: Healthcare-associated pneumonia HCAP is a relatively new category of pneumonia. It refers to infections that occur prior to hospital admission in patients with specific risk factors following contact or exposure to a healthcare environment. There is currently no scoring index to predict the outcomes of HCAP patients. We applied and compared different community acquired pneumonia CAP scoring indices to predict 30-day mortality and 3-day and 14-day intensive care unit ICU admission in patients with HCAP. Methods: We conducted a retrospective cohort study based on an inpatient database from six medical centers, recruiting a total of 444 patients with HCAP between 1 January 2007 and 31 December 2007. Pneumonia severity scoring indices including PSI pneumonia severity index, CURB 65 confusion, urea, respiratory rate, blood pressure , age 65, IDSA/ATS Infectious Diseases Society of America/American Thoracic Society, modified ATS rule, SCAP severe community acquired pneumonia, SMART-COP systolic blood pressure, multilobar involvement, albumin, respiratory rate, tachycardia, confusion, oxygenation, pH, SMRT- CO systolic blood pressure, multilobar involvement, respiratory rate, tachycardia, confusion, oxygenation, and SOAR systolic blood pressure, oxygenation, age, respiratory rate were calculated for each patient. Patient characteristics, co-morbidities, pneumonia pathogen culture results, length of hospital stay LOS, and length of ICU stay were also recorded. Results: PSI > 90 has the highest sensitivity in predicting mortality, followed by CURB-65 >= 2 and SCAP > 9 SCAP score area under the curve AUC: 0.71, PSI AUC: 0.70 and CURB-65 AUC: 0.66. Compared to PSI, modified ATS, IDSA/ATS, SCAP, and SMART-COP were easy to calculate. For predicting ICU admission Day 3 and Day 14, modified ATS AUC: 0.84, 0.82 , SMART-COP AUC: 0.84, 0.82, SCAP AUC: 0.82, 0.80 and IDSA/ ATS AUC: 0.80, 0 .79 performed better statistically significant difference than PSI, CURB- 65, SOAR and SMRT-CO. Conclusions: The utility of the scoring indices for risk assessment in patients with healthcare-associated pneumonia shows that the scoring indices originally designed for CAP can be applied to HCAP

Partial Nephrectomy in the Treatment of Localized Renal Cell Carcinoma — Experience of Taichung Veterans General Hospital

BackgroundPartial nephrectomy has been considered an effective and efficient method in the treatment of localized renal cell carcinoma. Herein, we retrospectively review our experience with partial nephrectomy in the treatment of localized renal cell carcinoma and compared it with patients who received radical nephrectomy.MethodsFrom 1982 to 2005, 35 patients who received partial nephrectomy for localized renal cell carcinoma were enrolled in this study. Ten patients were female (28.6%). The median age was 70 years (range, 42–82 years). Sixteen (45.7%) patients had pathologic T1a tumors; 17 (48.6%) patients had pathologic T1b tumors and 2 (5.7%) patients had pathologic T2 tumor (7 cm). In the meantime, 128 patients who had T1N0M0 renal cell carcinoma and who received radical nephrectomy were assigned to a control group. Thirty-nine patients (30.5%) were female in this group. The median age was 62 years (range, 30–83 years). The tumor characteristics, location, surgical techniques and patient survival were subsequently compared.ResultsThe median tumor size in the partial nephrectomy group was 3.9 cm (range, 1.5–7.0 cm), and it was 4.5 cm (range, 1–6.5 cm) in radical nephrectomy group. The tumor size was smaller in the partial nephrectomy group (p = 0.003). The median follow-up period was 4.38 years (range, 0.05–17.99 years) in the partial nephrectomy group and 5.66 years (range, 0.01–22.25 years) in the radical nephrectomy group. There was no local recurrence or distant metastasis in the partial nephrectomy group. The 5-year overall survival was 85.0% compared with 91.4% in the radical nephrectomy group (p = 0.126). The 5-year disease-specific survival in the partial nephrectomy group was 100%. The postoperative serum creatinine level increased to > 2.0 mg/dL in 5 (14.3%) patients in the partial nephrectomy group, but no patient needed hemodialysis during follow-up.ConclusionFrom our review, partial nephrectomy is safe and provides excellent disease control in the treatment of localized renal cell carcinoma in selected patients. Renal function preservation was observed in the partial nephrectomy group, while the operated kidney showed functioning in the follow-up nuclear medicine survey

Functional interaction between Env oncogene from Jaagsiekte sheep retrovirus and tumor suppressor Sprouty2

<p>Abstract</p> <p>Background</p> <p>Jaagsiekte sheep retrovirus (JSRV) is a type D retrovirus capable of transforming target cells <it>in vitro </it>and <it>in vivo</it>. The Envelope <it>(Env) </it>gene from JSRV and from related retroviruses can induce oncogenic transformation, although the detailed mechanism is yet to be clearly understood. Host cell factors are envisaged to play a critical determining role in the regulation of <it>Env</it>-mediated cell transformation.</p> <p>Results</p> <p>JSRV <it>Env</it>-mediated transformation of a lung adenocarcinoma cell line induced rapid proliferation, anchorage-independent growth and tumor formation, but completely abrogated the migration ability. An analysis of the signaling scenario in the transformed cells suggested the involvement of the ERK pathway regulated by Sprouty2 in cell migration, and the PI3K-Akt and STAT3 pathways in proliferation and anchorage-independence. On the other hand, in a normal lung epithelial cell line, <it>Env</it>-mediated transformation only decreased the migration potential while the other functions remained unaltered. We observed that <it>Env </it>induced the expression of a tumor suppressor, Sprouty2, suggesting a correlation between <it>Env</it>-effect and Sprouty2 expression. Overexpression of Sprouty2 <it>per se </it>not only decreased the migratory potential and tumor formation potential of the target cells but also made them resistant to subsequent <it>Env</it>-mediated transformation. On the other hand, over expression of the functional mutants of Sprouty2 had no inhibitory effect, confirming the role of Sprouty2 as a tumor suppressor.</p> <p>Conclusions</p> <p>Our studies demonstrate that <it>Env </it>and Sprouty2 have a functional relationship, probably through shared signaling network. Sprouty2 functions as a tumor suppressor regulating oncogenic transformation of cells, and it therefore has the potential to be exploited as a therapeutic anti-cancer agent.</p