Process engineering of biopharmaceutical production in moss bioreactors via model-based description and evaluation of phytohormone impact

The moss Physcomitrella is an interesting production host for recombinant biopharmaceuticals. Here we produced MFHR1, a synthetic complement regulator which has been proposed for the treatment of diseases associated to the complement system as part of human innate immunity. We studied the impact of different operation modes for the production process in 5 L stirred-tank photobioreactors. The total amount of recombinant protein was doubled by using fed-batch or batch compared to semi-continuous operation, although the maximum specific productivity (mg MFHR1/g FW) increased just by 35%. We proposed an unstructured kinetic model which fits accurately with the experimental data in batch and semi-continuous operation under autotrophic conditions with 2% CO2 enrichment. The model is able to predict recombinant protein production, nitrate uptake and biomass growth, which is useful for process control and optimization. We investigated strategies to further increase MFHR1 production. While mixotrophic and heterotrophic conditions decreased the MFHR1-specific productivity compared to autotrophic conditions, addition of the phytohormone auxin (NAA, 10 μM) to the medium enhanced it by 470% in shaken flasks and up to 230% and 260%, in batch and fed-batch bioreactors, respectively. Supporting this finding, the auxin-synthesis inhibitor L-Kynurenine (100 μM) decreased MFHR1 production significantly by 110% and 580% at day 7 and 18, respectively. Expression analysis revealed that the MFHR1 transgene, driven by the Physcomitrella actin5 (PpAct5) promoter, was upregulated 16 hours after NAA addition and remained enhanced over the whole process, whereas the auxin-responsive gene PpIAA1A was upregulated within the first two hours, indicating that the effect of auxin on PpAct5 promoter-driven expression is indirect. Furthermore, the day of NAA supplementation was crucial, leading to an up to 8-fold increase of MFHR1-specific productivity (0.82 mg MFHR1/ g fresh weight, 150 mg accumulated over 7 days) compared to the productivity reported previously. Our findings are likely to be applicable to other plant-based expression systems to increase biopharmaceutical production and yields.

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Novel Applications of Nanotechnology in Controlling HIV and HSV Infections

Current Drug Research Reviews ◽

10.2174/2589977512999201124121931 ◽

2020 ◽

Vol 12 ◽

Author(s):

Sai Akilesh M ◽

Ashish Wadhwani

Keyword(s):

Drug Delivery ◽

Viral Infections ◽

Polymeric Nanoparticles ◽

Volume Ratio ◽

Fold Increase ◽

Multi Drug Resistance ◽

Health Crisis ◽

Pharmaceutical Properties ◽

Simplex Virus ◽

The Impact

: Infectious diseases have been prevalent since many decades and viral pathogens have caused global health crisis and economic meltdown on a devastating scale. High occurrence of newer viral infections in the recent years, in spite of the progress achieved in the field of pharmaceutical sciences defines the critical need for newer and more effective antiviral therapies and diagnostics. The incidence of multi-drug resistance and adverse effects due to the prolonged use of anti-viral therapy is also a major concern. Nanotechnology offers a cutting edge platform for the development of novel compounds and formulations for biomedical applications. The unique properties of nano-based materials can be attributed to the multi-fold increase in the surface to volume ratio at the nano-scale, tunable surface properties of charge and chemical moieties. Idealistic pharmaceutical properties such as increased bioavailability and retention times, lower toxicity profiles, sustained release formulations, lower dosage forms and most importantly, targeted drug delivery can be achieved through the approach of nanotechnology. The extensively researched nano-based materials are metal and polymeric nanoparticles, dendrimers and micelles, nano-drug delivery vesicles, liposomes and lipid based nanoparticles. In this review article, the impact of nanotechnology on the treatment of Human Immunodeficiency Virus (HIV) and Herpes Simplex Virus (HSV) viral infections during the last decade are outlined.

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Administrative Coding Methods Impact Surgical Site Infection Rates

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2020.616 ◽

2020 ◽

Vol 41 (S1) ◽

pp. s111-s112

Author(s):

Mohammed Alsuhaibani ◽

Mohammed Alzunitan ◽

Kyle Jenn ◽

Daniel Diekema ◽

Michael Edmond ◽

...

Keyword(s):

Cesarean Section ◽

Surgical Procedures ◽

Hip Prosthesis ◽

Knee Prosthesis ◽

Surgical Site Infections ◽

Fold Increase ◽

Abdominal Hysterectomy ◽

Absolute Difference ◽

Coding Method ◽

The Impact

Background: Surveillance for surgical site infections (SSI) is recommended by the CDC. Currently, colon and abdominal hysterectomy SSI rates are publicly available and impact hospital reimbursement. However, the CDC NHSN allows surgical procedures to be abstracted based on International Classification of Diseases, Tenth Revision (ICD-10) or current procedural terminology (CPT) codes. We assessed the impact of using ICD and/or CPT codes on the number of cases abstracted and SSI rates. Methods: We retrieved administrative codes (ICD and/or CPT) for procedures performed at the University of Iowa Hospitals & Clinics over 1 year: October 2018–September 2019. We included 10 procedure types: colon, hysterectomy, cesarean section, breast, cardiac, craniotomy, spinal fusion, laminectomy, hip prosthesis, and knee prosthesis surgeries. We then calculated the number of procedures that would be abstracted if we used different permutations in administration codes: (1) ICD codes only, (2) CPT codes only, (3) both ICD and CPT codes, and (4) at least 1 code from either ICD or CPT. We then calculated the impact on SSI rates based on any of the 4 coding permutations. Results: In total, 9,583 surgical procedures and 180 SSIs were detected during the study period using the fourth method (ICD or CPT codes). Denominators varied according to procedure type and coding method used. The number of procedures abstracted for breast surgery had a >10-fold difference if reported based on ICD only versus ICD or CPT codes (104 vs 1,109). Hip prosthesis had the lowest variation (638 vs 767). For SSI rates, cesarean section showed almost a 3-fold increment (2.6% when using ICD only to 7.32% with both ICD & CPT), whereas abdominal hysterectomy showed nearly a 2-fold increase (1.14% when using CPT only to 2.22% with both ICD & CPT codes). However, SSI rates remained fairly similar for craniotomy (0.14% absolute difference), hip prosthesis (0.24% absolute difference), and colon (0.09% absolute difference) despite differences in the number of abstracted procedures and coding methods. Conclusions: Denominators and SSI rates vary depending on the coding method used. Variations in the number of procedures abstracted and their subsequent impact on SSI rates were not predictable. Variations in coding methods used by hospitals could impact interhospital comparisons and benchmarking, potentially leading to disparities in public reporting and hospital penalties.Funding: NoneDisclosures: None

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Combinations of Freeze-Dried Amorphous Vardenafil Hydrochloride with Saccharides as A Way to Enhance Dissolution Rate and Permeability

Pharmaceuticals ◽

10.3390/ph14050453 ◽

2021 ◽

Vol 14 (5) ◽

pp. 453

Author(s):

Gabriela Wiergowska ◽

Dominika Ludowicz ◽

Kamil Wdowiak ◽

Andrzej Miklaszewski ◽

Kornelia Lewandowska ◽

...

Keyword(s):

Physicochemical Properties ◽

Density Functional ◽

Hydroxypropyl Methylcellulose ◽

Fold Increase ◽

Crystalline Form ◽

Amorphous Form ◽

Freeze Dried ◽

Apparent Solubility ◽

Gastrointestinal Epithelium ◽

The Impact

To improve physicochemical properties of vardenafil hydrochloride (VAR), its amorphous form and combinations with excipients—hydroxypropyl methylcellulose (HPMC) and β-cyclodextrin (β-CD)—were prepared. The impact of the modification on physicochemical properties was estimated by comparing amorphous mixtures of VAR to their crystalline form. The amorphous form of VAR was obtained as a result of the freeze-drying process. Confirmation of the identity of the amorphous dispersion of VAR was obtained through the use of comprehensive analysis techniques—X-ray powder diffraction (PXRD) and differential scanning calorimetry (DSC), supported by FT-IR (Fourier-transform infrared spectroscopy) coupled with density functional theory (DFT) calculations. The amorphous mixtures of VAR increased its apparent solubility compared to the crystalline form. Moreover, a nearly 1.3-fold increase of amorphous VAR permeability through membranes simulating gastrointestinal epithelium as a consequence of the changes of apparent solubility (Papp crystalline VAR = 6.83 × 10−6 cm/s vs. Papp amorphous VAR = 8.75 × 10−6 cm/s) was observed, especially for its combinations with β-CD in the ratio of 1:5—more than 1.5-fold increase (Papp amorphous VAR = 8.75 × 10−6 cm/s vs. Papp amorphous VAR:β-CD 1:5 = 13.43 × 10−6 cm/s). The stability of the amorphous VAR was confirmed for 7 months. The HPMC and β-CD are effective modifiers of its apparent solubility and permeation through membranes simulating gastrointestinal epithelium, suggesting a possibility of a stronger pharmacological effect.

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Grill Workers Exposure to Polycyclic Aromatic Hydrocarbons: Levels and Excretion Profiles of the Urinary Biomarkers

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18010230 ◽

2020 ◽

Vol 18 (1) ◽

pp. 230

Author(s):

Marta Oliveira ◽

Sílvia Capelas ◽

Cristina Delerue-Matos ◽

Simone Morais

Keyword(s):

Polycyclic Aromatic Hydrocarbons ◽

Aromatic Hydrocarbons ◽

High Performance ◽

Principal Component ◽

Fold Increase ◽

Urinary Biomarkers ◽

Analysis Model ◽

Exposed Workers ◽

Polycyclic Aromatic ◽

The Impact

Grilling activities release large amounts of hazardous pollutants, but information on restaurant grill workers’ exposure to polycyclic aromatic hydrocarbons (PAHs) is almost inexistent. This study assessed the impact of grilling emissions on total workers’ exposure to PAHs by evaluating the concentrations of six urinary biomarkers of exposure (OHPAHs): naphthalene, acenaphthene, fluorene, phenanthrene, pyrene, and benzo(a)pyrene. Individual levels and excretion profiles of urinary OHPAHs were determined during working and nonworking periods. Urinary OHPAHs were quantified by high-performance liquid-chromatography with fluorescence detection. Levels of total OHPAHs (∑OHPAHs) were significantly increased (about nine times; p ≤ 0.001) during working comparatively with nonworking days. Urinary 1-hydroxynaphthalene + 1-hydroxyacenapthene and 2-hydroxyfluorene presented the highest increments (ca. 23- and 6-fold increase, respectively), followed by 1-hydroxyphenanthrene (ca. 2.3 times) and 1-hydroxypyrene (ca. 1.8 times). Additionally, 1-hydroxypyrene levels were higher than the benchmark, 0.5 µmol/mol creatinine, in 5% of exposed workers. Moreover, 3-hydroxybenzo(a)pyrene, biomarker of exposure to carcinogenic PAHs, was detected in 13% of exposed workers. Individual excretion profiles showed a cumulative increase in ∑OHPAHs during consecutive working days. A principal component analysis model partially discriminated workers’ exposure during working and nonworking periods showing the impact of grilling activities. Urinary OHPAHs were increased in grill workers during working days.

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Serotonin depletion reduces both acquisition and expression of context-conditioned fear

Acta Neuropsychiatrica ◽

10.1017/neu.2021.3 ◽

2021 ◽

pp. 1-8

Author(s):

S. Melker Hagsäter ◽

Robert Pettersson ◽

Axel Holmäng ◽

Elias Eriksson

Keyword(s):

Unconditioned Stimulus ◽

Memory Consolidation ◽

Clinical Studies ◽

Conditioned Fear ◽

Sprague Dawley ◽

Serotonin Synthesis ◽

Synthesis Inhibitor ◽

Serotonin Depletion ◽

Sprague Dawley Rats ◽

The Impact

Abstract Objective: Whereas numerous experimental and clinical studies suggest a complex involvement of serotonin in the regulation of anxiety, it remains to be clarified if the dominating impact of this transmitter is best described as anxiety-reducing or anxiety-promoting. The aim of this study was to assess the impact of serotonin depletion on acquisition, consolidation, and expression of conditioned fear. Methods: Male Sprague–Dawley rats were exposed to foot shocks as unconditioned stimulus and assessed with respect to freezing behaviour when re-subjected to context. Serotonin depletion was achieved by administration of a serotonin synthesis inhibitor, para-chlorophenylalanine (PCPA) (300 mg/kg daily × 3), (i) throughout the period from (and including) acquisition to (and including) expression, (ii) during acquisition but not expression, (iii) after acquisition only, and (iv) during expression only. Results: The time spent freezing was significantly reduced in animals that were serotonin-depleted during the entire period from (and including) acquisition to (and including) expression, as well as in those being serotonin-depleted during either acquisition only or expression only. In contrast, PCPA administrated immediately after acquisition, that is during memory consolidation, did not impact the expression of conditioned fear. Conclusion: Intact serotonergic neurotransmission is important for both acquisition and expression of context-conditioned fear.

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Chitosan Oligosaccharides Stimulate the Efficacy of Somatic Embryogenesis in Different Genotypes of the Liriodendron Hybrid

Forests ◽

10.3390/f12050557 ◽

2021 ◽

Vol 12 (5) ◽

pp. 557

Author(s):

Asif Ali ◽

Jiaji Zhang ◽

Minmin Zhou ◽

Tingting Chen ◽

Liaqat Shah ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Antagonistic Effect ◽

Growth Speed ◽

Chitosan Oligosaccharide ◽

Chitosan Oligosaccharides ◽

Germination Ability ◽

Auxin Synthesis ◽

Branch Development ◽

The Impact ◽

Seed Setting Rate

Liriodendron hybrid (L. chinense × L. tulipifera), an essential medium-sized tree generally famous for its timber, is also used as an ornamental and greenery tool in many places around the world. The Liriodendron hybrid (L. hybrid) tree goes through many hurdles to achieve its maximum strength and vigor, such as loss of habitat, vast genetic variation, and low seed setting rate. The establishment of an effective and well-organized somatic embryogenesis (S.E.) system could be used to overcome these obstacles, rather than the old-fashioned seed culture and organogenesis. This study is based on the impact of chitosan oligosaccharide (COS) and its role in the induction of S.E. on the callus of four genotypes of the L. hybrid. The optimal concentration of COS could enhance the momentum and effectiveness in S.E.’s mechanism, which further improves the growth rate of the L. hybrid tree’s plantlets. This study shows that COS has a prominent role in endogenous hormones like indole acetic acid (IAA), zeatin (Z.T.), and gibberellic acid (GA3). Furthermore, COS improves the growth development, growth speed, as well as the development situation of plant germination ability. COS can also regulate branch development and root growth, which could be linked to the antagonistic effect on growth factors to some extent or by affecting auxin synthesis and polar transport.

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CHANGES IN THE IMPACT ASSESSMENT FAMILY 2003–2014: IMPLICATIONS FOR CONSIDERING ACHIEVEMENTS, GAPS AND FUTURE DIRECTIONS

Journal of Environmental Assessment Policy and Management ◽

10.1142/s1464333215500039 ◽

2015 ◽

Vol 17 (01) ◽

pp. 1550003 ◽

Cited By ~ 13

Author(s):

FRANK VANCLAY

Keyword(s):

Risk Assessment ◽

Impact Assessment ◽

Social Impact ◽

Public Involvement ◽

Cost Benefit Analysis ◽

Cost Benefit ◽

Fold Increase ◽

Social Risk ◽

Proportional Change ◽

The Impact

Over 150 forms of impact assessment can be identified using Google searches, with several new forms appearing since 2003. Since then, the popularity of the various members of the impact assessment family has changed, partly in response to legislative and regulatory changes, and general trends in society. The information explosion and expansion of the internet has resulted in a 32 fold increase in the number of hits for "impact assessment", now over 12 million. The conventional methods most frequently mentioned in 2003 had relatively low proportional change over the last 10 years but remain amongst the most frequently mentioned in 2014: risk assessment, public participation, cost-benefit analysis, public involvement, environmental monitoring, and project evaluation. The terms with highest proportional change (i.e. the super-hot topics) were primarily social concerns, including: equality impact assessment, welfare impact assessment, mental health impact assessment, disability impact assessment, human impact assessment, social impact assessment, and social risk assessment. Other terms that had high proportional change included life cycle impact assessment. Information about the relative popularity of the various forms of impact assessment is used in this paper to discuss issues and trends in the broad field of impact assessment.

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Improving Fatigue Life of Riveted Joints by Rivet Hole Sizing

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.598.141 ◽

2014 ◽

Vol 598 ◽

pp. 141-146

Author(s):

Adam Lipski ◽

Zbigniew Lis

Keyword(s):

Fatigue Life ◽

Fold Increase ◽

Fatigue Tests ◽

Aviation Industry ◽

Lap Joint ◽

Riveted Joints ◽

Riveting Process ◽

The Difference ◽

Structural Connection ◽

The Impact

The aim of this paper is to assess the impact of the rivet hole sizing process on the fatigue life based on the example of the structural connections characteristic for riveted joints used in aviation industry. Test specimens reflected the structural connection consisting in a riveted lap joint of an airplane plating stiffened with a T-bar. Connected plates and the T-bar are made of D16CzATW aluminum alloy. 3 mm diameter oval head solid rivets for aviation-related purposes were made of PA24 aluminum. During fatigue tests, individual specimens with non-sized holes and with sized holes were subjected to uniaxial, one-sided, fixed-amplitude loading (R = 0). It can be concluded from the fatigue life comparison that introduction of an additional operation in the riveting process, i.e. the hole sizing, results in significant, about two-fold increase of the fatigue life of the riveted structural connection, even at slight sizing degree. The difference of the specimen damage nature was observed between specimens with sized and non-sized holes.

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