Economic simulation of batch and continuous aqueous two-phase purification for viral products.

Vaccine manufacturing strategies that lower capital and production costs could improve vaccine access by reducing the cost per dose and encouraging localized manufacturing. Continuous processing is increasingly utilized to drive lower costs in biological manufacturing by requiring fewer capital and operating resources. Aqueous two-phase systems (ATPS) are a liquid-liquid extraction technique that enables continuous processing for viral vectors. To date, no economic comparison between viral vector purifications using traditional methods and ATPS has been published. In this work, economic simulations of traditional chromatography-based virus purification were compared to ATPS-based virus purification for the same product output in both batch and continuous modes. First, the modeling strategy was validated by re-creating a viral subunit manufacturing economic simulation. Then, ATPS capital and operating costs were compared to that of a traditional chromatography purification at multiple scales. At all scales, ATPS purification required less than 10% of the capital expenditure compared to chromatography-based purification. At an 11 kg per year production scale, the ATPS production costs were 50% less than purification with chromatography. Other chromatography configurations were explored, and may provide a production cost benefit to ATPS, but the purity and recovery were not experimentally verified. Batch and continuous ATPS were similar in capital and production costs. However, manual price adjustments suggest that continuous ATPS plant-building costs could be less than half that of batch ATPS at the 11 kg per year production scale. These simulations show the significant reduction in manufacturing costs that ATPS-based purification could deliver to the vaccine industry.

Role of Abdominal Ultrasound and Magnetic Resonance Imaging in Pregnant Women Presenting With Acute Abdominal Pain.

Background and Purpose: Abdominal pain during pregnancy is quite common; however, surgical pathology such as acute appendicitis as a cause of such pain is not. Diagnostic tests used in addition to history and physical examination (PE) are ultrasound (US) and magnetic resonance imaging (MRI). We elected to find the role of these tests in pregnant patients who presented to our emergency department with acute abdominal pain.Materials And Methods: Retrospective analysis of 136 pregnant women with acute abdominal pain presented to the emergency department (ED). We reviewed PE, US, MRI, gestational age, comorbid conditions, and length of stay. Statistical analysis was done using student's t-test and chi-square test. Institutional review board approval was obtained.Results: Mean age was 26 (±4.6) and the mean gestational age was 24 (±9.9) weeks. Of those patients, there were 81 patients who had an US and MRI performed. The US was positive in 16 patients, while the MRI was positive in 25 patients. Three patients went for appendectomy. The US sensitivity was 0% and specificity 79%. Positive predictive value for US was 0% and negative predictive value was 95%, which was less than 100%. The MRI likelihood ratios were calculated for each test's clinical application and demonstrated that the US test result was indeterminate for ruling in and for ruling out appendicitis while the MRI allowed for high ability to rule out the disease.Conclusion: In pregnant women with acute abdominal pain and a positive PE highly suggestive of surgical pathology, US had limited value and patients should proceed to MRI.

Traumatic brain injury in the elderly with high Glasgow coma scale and low injury severity scores: Factors influencing outcomes.

BACKGROUND: Current trauma activation guidelines do not clearly address age as a risk factor when leveling trauma patients. Glasgow coma scale (GCS) and mode of injury play a major role in leveling trauma patients. We studied the above relationship in our elderly patients presenting with traumatic head injury. METHODS: This study was a retrospective analysis of patients who presented to the emergency department with traumatic brain injuries. We classified the 270 patients into two groups. Group A was 64 years and younger, and group B was 65 years and older. Their GCS, ISS, age, sex, comorbidities, and anticoagulant use were abstracted. The primary outcome was mortality and length of stay. The groups were compared using an independent student's t-test and Chi-square analysis. The Cox regression analysis was used to analyze differences in the outcome while adjusting for the above factors. RESULTS: There were 140 patients in group A, and 130 patients in group B who presented to the ED with a GCS of 14-15 and an ISS of below 15. The mean ISS significantly differed between group A (6.2 ± 6.8) vs (7.9 ± 3.2) in group B (p < 0.0001). The most common diagnosis in group A was concussion (57.3%), while in group B was subdural and subarachnoid hemorrhage (55%). In group B, 13.8% presented as a level one or level two trauma activation. The mean hospital and intensive care stay for group A was 2.1 (±1.9) days and 0.9 (±1.32) days, respectively, versus 4.2 (±3.04) days and 2.4 (±2.02 days) for the elderly group B. Mortality in group A was zero and in group B was 3.8%. Cox regression analysis showed age as an independent predictor of death as well as length of stay. CONCLUSION: Elderly traumatic brain injury patients presenting to the ED with minor trauma and high GCS should be triaged at a higher level in most cases.

Hydroxyl Radical Generation Through the Fenton-Like Reaction of Hematin- and Catechol-Functionalized Microgels.

Hydroxyl radical (•OH) is a potent reactive oxygen species with the ability to degrade hazardous organic compounds, kill bacteria, and inactivate viruses. However, an off-the-shelf, portable, and easily activated biomaterial for generating •OH does not exist. Here, microgels were functionalized with catechol, an adhesive moiety found in mussel adhesive proteins, and hematin (HEM), a hydroxylated Fe3+ ion-containing porphyrin derivative. When the microgel was hydrated in an aqueous solution with physiological pH, molecular oxygen in the solution oxidized catechol to generate H2O2, which was further converted to •OH by HEM. The generated •OH was able to degrade organic dyes, including orange II and malachite green. Additionally, the generated •OH was antimicrobial against both gram-negative (Escherichia coli) and gram-positive (Staphylococcus epidermidis) bacteria with the initial concentration of 106-107 CFU/mL. These microgels also reduced the infectivity of a non-enveloped porcine parvovirus and an enveloped bovine viral diarrhea virus by 3.5 and 4.5 log reduction values, respectively (99.97-99.997% reduction in infectivity). These microgels were also functionalized with positively charged [2-(methacryloyloxy)ethyl] trimethylammonium chloride (METAC), which significantly enhanced the antibacterial and antiviral activities through electrostatic interaction between the negatively charged pathogens and the microgel. These microgels can potentially serve as a lightweight and portable source of disinfectant, for an on-demand generation of •OH with a wide range of applications.

Porcine parvovirus flocculation and removal in the presence of osmolytes.

Viruses can be modified into viral vaccines or gene therapy vectors in order to treat acquired or genetic diseases. To satisfy the current market demand, an improvement in current vaccine manufacturing is needed. Chromatography and nanofiltration are not suitable for all types of viruses. In this study, we propose to use virus flocculation with osmolytes, followed by microfiltration, as a potential virus purification process. We hypothesize that osmolytes strongly bind to water, thus leading to the formation of a hydration layer around the virus particles and stimulation of aggregation. We have discovered that osmolytes, including sugars, sugar alcohols and amino acids, preferentially flocculate porcine parvovirus (PPV), and demonstrate a >80% removal with a 0.2 µm filter while leaving model proteins in solution. This large pore size filter increases the flux and decreases the transmembrane pressure of typical virus filters. The best flocculants were tested for their ability to aggregate PPV at different concentrations, shear stress, pH and ionic strength. We were able to remove 96% of PPV in 3.0M glycine at a pH of 5. Glycine is also an excipient, and therefore may not require removal later in the process. Virus flocculation using osmolytes, followed by microfiltration could be used as an integrated process for virus purification.

A reappraisal of the clinical spectrum of North Carolina macular dystrophy.

PURPOSE: To characterize the clinical phenotypes and genotype of a large family with North Carolina macular dystrophy (NCMD). DESIGN: Observational, retrospective case series. PARTICIPANTS: Thirteen participants who were at risk of inheriting a dominantly transmitted disease gene from a 4-generation family from Baltimore were examined. METHODS: Thirteen participants underwent ophthalmic examination and genomic linkage analysis. Fundus photography, spectral-domain optical coherence tomography (SD-OCT), fluorescein angiography, ultrasonography, full-field electroretinography, and electro-oculography were performed on some patients. MAIN OUTCOME MEASURES: Description of clinical phenotypes with genomic linkage to the MCDR1 locus. RESULTS: Nine of 13 participants were affected with NCMD. There are variable and previously unreported clinical manifestations among affected individuals with NCMD, including drusen, macular staphyloma, choroidal neovascularization, a retinal pigment epithelial tear, and geographic atrophy. The distinctive and virtually pathognomonic grade 3 lesions in NCMD are neither staphylomas nor colobomas, as previously thought. As shown by ultrasonography and SD-OCT, they are deep chorioretinal excavations not involving the sclera, for which the authors propose a new term: macular caldera. Linkage analysis was performed, and the disease-causing gene in this family was mapped to the MCDR1 locus. CONCLUSIONS: North Carolina macular dystrophy has a wide spectrum of clinical phenotypes that resemble age-related macular degeneration except for their early age of onset.

Profiles of pain in Mississippi: results from the Southern Pain Prevalence Study.

OBJECTIVES: To estimate the prevalence, intensity, frequency, and origins of pain in Mississippi residents and explore the social and cultural aspects of pain and pain management by measuring the impacts of pain on everyday life as well as respondents' beliefs and attitudes toward pain. STUDY DESIGN: Cross-sectional survey of Mississippi adults. SUMMARY OF BACKGROUND DATA: Data from previous population surveys and clinical studies indicate that pain is a pervasive part of everyday life. The prevalence, intensity and frequency of reported pain is substantial throughout these studies. The literature documents large and profound consequences of pain both for individual lives and for society. The data also suggest that beliefs and attitudes rooted in the social and cultural context of society may help explain how pain is managed or endured. METHODS: Telephone interviews with a representative sample (random digit dialing) of 604 Mississippi adults were conducted utilizing a computer-assisted telephone interviewing system. The cooperation rate was 94.8% (5.2 % refusal). The Code of Standards and Ethics for Survey Research rate was 67.4% and the maximum sampling error was +/- 4.0% (95% confidence interval). The Mississippi survey data represent a subset of the data obtained in the six-state Southern Pain Prevalence Study. RESULTS: The pain prevalence rate was estimated to be 37% of the overall Mississippi sample: 9% of the sample reported severe pain on at least a monthly basis; 16% reported moderate pain; and 12% reported mild pain. Among those reporting pain, a majority (52%) experienced pain on a daily basis. The most commonly reported origins of pain were back pain (49%), leg and knee pain (41%), and shoulder.and arm pain (20%). Respondents also reported that both moderate and severe pain had substantial negative impacts on multiple facets of everyday life, including interference with sleep (84%), recreational/leisure activities (78%), ability to work (68%), sexual relations (43%), and relationships with others (36%). Mental health impacts for respondents with moderate to severe pain on at least a monthly basis included increased feelings of anxiety (66%), self-reported depression (63%), and loneliness (46%). When responding to questions regarding beliefs and attitudes about pain and pain management, the majority of respondents (62%) considered pain to be a normal part of everyday life. Many respondents felt that medicine should be saved until the pain becomes worse (55%), and a substantial number of respondents felt that good patients do not complain about pain to their doctors (22%). CONCLUSIONS: This study expands the body of knowledge about the prevalence of pain in Mississippi, suggesting that approximately one-third of the state's adults are affected by pain on at least a monthly basis. Most of this pain is moderate to severe and quite frequent--occurring daily for the majority of pain sufferers. The study also illuminates social and cultural dimensions of pain, revealing that a) the presence of pain negatively affects almost every facet of life, from sleep and work to relationships, leisure activities, and mental health, and b) respondents attitudes and beliefs are often at variance with modern approaches to pain management. Knowledge gained could have critical implications for understanding patients and the treatment of pain.