Process analytical technology (PAT) tools for the cultivation step in biopharmaceutical production

The process analytical technology (PAT) initiative is now 10 years old. This has resulted in the development of many tools and software packages dedicated to PAT application on pharmaceutical processes. However, most applications are restricted to small molecule drugs, mainly for the relatively simple process steps like drying or tableting where only a limited number of parameters need to be controlled. A big challenge for PAT still lies in applications for biopharmaceuticals and then especially in the cultivation process step, where the quality of a biopharmaceutical product is largely determined. This review gives an overview of the currently available tools for monitoring and controlling the biopharmaceutical cultivation step and of the main challenges for the most common cell platforms (i.e. Escherichia coli, yeast, and mammalian cells) used in biopharmaceutical manufacturing. The real challenge is to understand how intracellular mechanisms (from synthesis to excretion) influence the quality of biopharmaceuticals and how these mechanisms can be monitored and controlled to yield the desired end product quality. Modern “omics” tools and advanced process analyzers have opened up the way for PAT applications for the biopharmaceutical cultivation process step.     

Hawai‘i’s Mountain-to-Sea Ecosystems: Social–Ecological Microcosms for Sustainability Science and Practice

There is an urgent need to develop the underlying theory and principles of “sustainability science,” based on an understanding of the fundamental interactions between nature and humans. This requires a new research and education paradigm that embraces biocomplexity, integrates the physical, biological, and social sciences, and uses a coupled, human–natural systems approach. An initiative aligned with this paradigm and approach, and centered on the Hawaiian Island’s unique mountain-to-sea ecosystems, is developing at the University of Hawai‘i. These ecosystems, extending from upland tropical forests to the fringing coral reefs, correspond to the roughly wedge-shaped catchments, traditionally called ahupua‘a in the Hawaiian language. Despite the collapse of the ahupua‘a system and, tragically, the Native Hawaiian population, its legacy of ecological and cultural stewardship remains. This legacy, and the potential of these ecosystems as microcosms for addressing the core questions of sustainability science, has provided the impetus for a growing number of projects employing a social–ecological systems perspective. An overview of three projects that employ a “learning community” approach and cultural stewardship perspective inspired by the ahupua‘a system is provided. These include the Ecosystems Thrust Area of Hawai‘i EPSCoR, a U.S. National Science Foundation research infrastructure program, focused on ecosystem research and monitoring activities; a sustainability curriculum program, Mālama I Ka ‘Āina, of the College of Education; and a project that builds on programs of the Division of Ecology and Health and its affiliated Asia-Pacific Center for Infectious Disease Ecology, linking ecosystem resilience and infectious diseases.     

Catheter-based impedance measurements in the right atrium for continuously monitoring hematocrit and estimating blood viscosity changes; an in vivo feasibility study in swine

Hematocrit is the most important determinant of whole blood viscosity and it affects thrombosis. As hematocrit can be measured accurately in vitro by using an electrical impedance technique, aim of the present study is to investigate the diagnostic potential of using this technique in vivo to continuously monitor hematocrit.

Characteristics of a special catheter for in vivo measurement of electrical resistivity in blood in the right atrium are described. In five anesthetized swine hematocrit is monitored continuously with this catheter while different levels of hemoconcentration are induced. In addition, blood viscosity is increased by inducing ‘acute phase’ reaction the day before surgery, resulting in variable degree of elevated fibrinogen levels in the five swine. Good reproducibility of the resistivity measurements (S.D.<0.01) and excellent correlation between resistivity data in vivo and hematocrit levels in each swine are found (r²=0.95–0.99). Furthermore, stepwise regression analysis of data from all swine shows a highly significant contribution also of other important parameters of blood viscosity, such as fibrinogen, total protein and temperature (cumulative r²=0.97).

Determining hematocrit continuously in vivo by electrical resistivity measurements with a catheter in the right atrium is feasible and these measurements correlate significantly also with other important parameters of blood viscosity.     

Continuous toxicity monitoring in phase II trials in oncology

The goal of a phase II trial in oncology is to evaluate the efficacy of a new therapy. The dose investigated in a phase II trial is usually an estimate of a maximum-tolerated dose obtained in a preceding phase I trial. Because this estimate is imprecise, stopping rules for toxicity are used in many phase II trials. We give recommendations on how to construct stopping rules to monitor toxicity continuously. A table is provided from which Pocock stopping boundaries can be easily obtained for a range of toxicity rates and sample sizes. Estimation of the probability of toxicity and response is also discussed.     

Ragweed in the Czech Republic

During the last years, a well documented expansion ofragweed (Ambrosia artemisiifolia L., Ambrosia trifida L.) over the Mediterranean andtemperate Europe has been in progress. The currentdistribution of ragweed plants in the Czech Republicis summarized and the ragweed pollen concentration asmonitored by 12 pollen stations in the country isdiscussed. The present situation in the ragweed pollensensitization among children and adults with pollenallergy in Brno is described. So far no dangerousexpansion of ragweed plants in our country has beenobserved. Ragweed pollen concentration is occasionallysignificant in the Brno station only, other pollenstations are reporting insignificant amounts ofragweed pollen during August-September periods,although there has been a steady increase in ragweedpollen concentration in the Prague area over the lastfive years. Skin prick tests and/or specific IgEmeasurements with ragweed allergen were performed on94 children with pollen allergy in the Brno region in1995 and on 206, 210 and 229 adult allergic patientsin 1995, 1996 and 1997, respectively. Positive skinreaction or positive specific IgE to ragweed was foundin 22% children and in 25% (1995), 19% (1996) and 25% (1997) adults with pollen allergy. It isconcluded that ragweed does not seem to represent anyimminent major threat to the allergic population inthe Czech Republic until now, however, it remains apotentially very dangerous allergen.     

Rapid monitoring of microbial contamination on herbal medicines by fluorescent staining method

AIMS: To apply fluorescent staining method for fast assessment of microbial quality of herbal medicines. METHODS AND RESULTS: The number of total bacteria and esterase-active bacteria on powdered traditional Chinese medicines were enumerated by fluorescent staining method using 6-carboxyfluorescein diacetate (6CFDA) and 4',6-diamidino-2-phenylindole (DAPI), and they were compared with colony-forming units (CFU). The CFU was approximately 10(3) per gram in ginseng radix, and no bacterial colonies were detected from others. However, the total bacterial number (TDC) was more than 10(7) per gram, and number of bacteria possessing esterase activity ranged from 1 to 3% of TDC. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Many bacteria in each Chinese medicine had enzyme activity and most of them could not be detected by conventional plate counting technique. Enumeration of bacterial cells on traditional Chinese medicines by fluorescent staining method requires less than 1 h. The double staining method with 6CFDA and DAPI could be applicable to rapid microbial monitoring of crude drugs.     

How Can a Patient Blind to Radial Motion Discriminate Shifts in the Center-of-Motion?

Within biologically constrained models of heading and complex motion processing, localization of the center-of-motion (COM) is typically an implicit property arising from the precise computation of radial motion direction associated with an observers forward self-motion. In the work presented here we report psychophysical data from a motion-impaired stroke patient, GZ, whose pattern of visual motion deficits is inconsistent with this view. We show that while GZ is able to discriminate direction in circular motions she is unable to discriminate direction in radial motion patterns. GZs inability to discriminate radial motion is in stark contrast with her ability to localize the COM in such stimuli and suggests that recovery of the COM does not necessarily require an explicit representation of radial motion direction. We propose that this dichotomy can be explained by a circular template mechanism that minimizes a global motion error relative to the visual motion input, and we demonstrate that a sparse population of such templates is computationally sufficient to account for human psychophysical performance in general and in particular, explains GZs performance. Recent re-analysis of the predicted receptive field structures in several existing heading models provides additional support for this type of circular template mechanism and suggests the human visual system may have available circular motion mechanisms for heading estimation.