Plant Milking Technology—An Innovative and Sustainable Process to Produce Highly Active Extracts from Plant Roots

We have used an original technology (Plant Milking Technology) based on aeroponic cultivation of plants associated with the gentle recovery of active ingredients from roots. Extraction of bioactive molecules was achieved by soaking the roots, still attached to the living plants, into a nontoxic solvent for a 2 h period. This nondestructive recovery process allows using the same root biomass for successive harvesting dates, in a recyclable way. We have applied this technology to Morus alba L. (mulberry tree), an emblematic tree of the Traditional Chinese Medicine (TCM). Trees were aeroponically grown in large-scale devices (100 m2) and were submitted to nitrogen deprivation to increase the content in active molecules (prenylated flavonoids). The Plant Milking technology applied to Morus alba L. allowed to produce an extract enriched in prenylated compounds (18-fold increase when compared to commercial root extract). Prenylated flavonoids (moracenin A and B, kuwanon C, wittiorumin F, morusin) presented a high affinity for the aged-associated collagenase enzyme, which was confirmed by activity inhibition. In accordance, M. alba extract presents efficient properties to regulate the skin matrisome, which is critical during skin aging. The benefits have been especially confirmed in vivo on wrinkle reduction, in a clinical study that involved aged women. Plant Milking technology is an optimal solution to produce active ingredients from plant roots, including trees, that meet both customer expectations around sustainability, as well as the need for an efficient production system for biotechnologists.

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Monobac System–A Single Baculovirus for the Production of rAAV

Microorganisms ◽

10.3390/microorganisms9091799 ◽

2021 ◽

Vol 9 (9) ◽

pp. 1799

Author(s):

Lionel Galibert ◽

Aurélien Jacob ◽

Adrien Savy ◽

Yohann Dickx ◽

Delphine Bonnin ◽

...

Keyword(s):

Large Scale ◽

Viral Vector ◽

Cell System ◽

Lower Amount ◽

Efficient Production ◽

Baculovirus Genome ◽

Raav Vector ◽

System A ◽

Raav Production

Large-scale manufacturing of rAAV is a bottleneck for the development of genetic disease treatments. The baculovirus/Sf9 cell system underpins the first rAAV treatment approved by EMA and remains one of the most advanced platforms for rAAV manufacturing. Despite early successes, rAAV is still a complex biomaterial to produce. Efficient production of the recombinant viral vector requires that AAV replicase and capsid genes be co-located with the recombinant AAV genome. Here, we present the Monobac system, a singular, modified baculovirus genome that contains all of these functions. To assess the relative yields between the dual baculovirus and Monobac systems, we prepared each system with a transgene encoding γSGC and evaluated vectors’ potency in vivo. Our results show that rAAV production using the Monobac system not only yields higher titers of rAAV vector but also a lower amount of DNA contamination from baculovirus.

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Efficient large-scale production and concentration of HIV-1-based lentiviral vectors for use in vivo

Physiological Genomics ◽

10.1152/physiolgenomics.00135.2002 ◽

2003 ◽

Vol 12 (3) ◽

pp. 221-228 ◽

Cited By ~ 120

Author(s):

Jason E. Coleman ◽

Matthew J. Huentelman ◽

Sergey Kasparov ◽

Beverly L. Metcalfe ◽

Julian F. R. Paton ◽

...

Keyword(s):

Animal Studies ◽

Large Scale ◽

Lentiviral Vector ◽

High Titer ◽

Lentiviral Vectors ◽

Efficient Production ◽

Scale Production ◽

Peripheral Tissues ◽

Large Scale Production

The aim of this study was to develop an efficient method for packaging and concentrating lentiviral vectors that consistently yields high-titer virus on a scale suitable for in vivo applications. Transient cotransfection of 293T packaging cells with DNA plasmids encoding lentiviral vector components was optimized using SuperFect, an activated dendrimer-based transfection reagent. The use of SuperFect allowed reproducible and efficient production of high-titer lentiviral vector at concentrations greater than 1 × 107transducing units per ml (TU/ml) and required less than one-third of the total amount of DNA used in traditional calcium phosphate transfection methods. Viral titers were further increased using a novel concentration protocol that yielded an average final titer of 1.4 × 1010TU/ml. Lentiviruses produced using these methods exhibited efficient transduction of central nervous system and peripheral tissues in vivo. The method is reproducible and can be scaled up to facilitate the use of these vectors in animal studies.

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Valorization of Wine-Making By-Products’ Extracts in Cosmetics

Cosmetics ◽

10.3390/cosmetics8040109 ◽

2021 ◽

Vol 8 (4) ◽

pp. 109

Author(s):

Israa Hoss ◽

Hiba N. Rajha ◽

Rindala El Khoury ◽

Sahar Youssef ◽

Maria Letizia Manca ◽

...

Keyword(s):

Phenolic Compounds ◽

Value Chain ◽

Grape Pomace ◽

Industrial Wastes ◽

Bioactive Molecules ◽

Cosmetic Products ◽

Active Ingredients ◽

By Products

The increased demand for conscious, sustainable and beneficial products by the consumers has pushed researchers from both industries and universities worldwide to search for smart strategies capable of reducing the environmental footprint, especially the ones connected with industrial wastes. Among various by-products, generally considered as waste, those obtained by winemaking industries have attracted the attention of a wide variety of companies, other than the vineries. In particular, grape pomaces are considered of interest due to their high content in bioactive molecules, especially phenolic compounds. The latter can be recovered from grape pomace and used as active ingredients in easily marketable cosmetic products. Indeed, phenolic compounds are well known for their remarkable beneficial properties at the skin level, such as antioxidant, antiaging, anti-hyperpigmentation and photoprotective effects. The exploitation of the bioactives contained in grape pomaces to obtain high value cosmetics may support the growing of innovative start-ups and expand the value chain of grapes. This review aims to describe the strategies for recovery of polyphenols from grape pomace, to highlight the beneficial potential of these extracts, both in vitro and in vivo, and their potential utilization as active ingredients in cosmetic products.

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Purification of the Antihemophilic Factor by Gel Filtration on Agarose

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651394 ◽

1969 ◽

Vol 22 (03) ◽

pp. 577-583 ◽

Cited By ~ 3

Author(s):

M.M.P Paulssen ◽

A.C.M.G.B Wouterlood ◽

H.L.M.A Scheffers

Keyword(s):

Factor Viii ◽

Plasma Proteins ◽

Large Scale ◽

Gel Filtration ◽

Large Scale Preparation ◽

Scale Preparation ◽

Antihemophilic Factor

SummaryFactor VIII can be isolated from plasma proteins, including fibrinogen by chromatography on agarose. The best results were obtained with Sepharose 6B. Large scale preparation is also possible when cryoprecipitate is separated by chromatography. In most fractions containing factor VIII a turbidity is observed which may be due to the presence of chylomicrons.The purified factor VIII was active in vivo as well as in vitro.

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The Effect of Active Site-inhibited Factor VIIa on Tissue Factor-initiated Coagulation Using Platelets before and after Aspirin Administration

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657715 ◽

1997 ◽

Vol 78 (04) ◽

pp. 1202-1208 ◽

Cited By ~ 18

Author(s):

Marianne Kjalke ◽

Julie A Oliver ◽

Dougald M Monroe ◽

Maureane Hoffman ◽

Mirella Ezban ◽

...

Keyword(s):

Tissue Factor ◽

Active Site ◽

Large Scale ◽

Thrombin Generation ◽

Factor Viia ◽

Dependent Manner ◽

Antithrombotic Agent ◽

Before And After ◽

Ic50 Values

SummaryActive site-inactivated factor VIIa has potential as an antithrombotic agent. The effects of D-Phe-L-Phe-L-Arg-chloromethyl ketone-treated factor VIla (FFR-FVIIa) were evaluated in a cell-based system mimicking in vivo initiation of coagulation. FFR-FVIIa inhibited platelet activation (as measured by expression of P-selectin) and subsequent large-scale thrombin generation in a dose-dependent manner with IC50 values of 1.4 ± 0.8 nM (n = 8) and 0.9 ± 0.7 nM (n = 7), respectively. Kd for factor VIIa binding to monocytes ki for FFR-FVIIa competing with factor VIIa were similar (11.4 ± 0.8 pM and 10.6 ± 1.1 pM, respectively), showing that FFR-FVIIa binds to tissue factor in the tenase complex with the same affinity as factor VIIa. Using platelets from volunteers before and after ingestion of aspirin (1.3 g), there were no significant differences in the IC50 values of FFR-FVIIa [after aspirin ingestion, the IC50 values were 1.7 ± 0.9 nM (n = 8) for P-selectin expression, p = 0.37, and 1.4 ± 1.3 nM (n = 7) for thrombin generation, p = 0.38]. This shows that aspirin treatment of platelets does not influence the inhibition of tissue factor-initiated coagulation by FFR-FVIIa, probably because thrombin activation of platelets is not entirely dependent upon expression of thromboxane A2.

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Acute and chronic toxicity studies on Polygonum glabrum in experimental animals

International Journal of Pharmacometrics and Integrated Biosciences ◽

10.26452/ijpib.v3i1.1216 ◽

2020 ◽

Vol 3 (1) ◽

pp. 7-10

Author(s):

Saravanakumar A ◽

Gandhimathi R

Keyword(s):

Chronic Toxicity ◽

Clinical Stage ◽

Herbal Medicines ◽

Plant Roots ◽

Root Extract ◽

Experimental Animals ◽

Toxicity Studies ◽

Acute And Chronic Toxicity ◽

Diuretic Agents

Polygonum glabrum is being used in traditional and folklore medicine to treat pneumonia and jaundice. Plant roots are used in ayurvedic preparations to treat fever and colic. The leaves are used as diuretic agents and process vermifuge action. Plant decoction is also used in the treatment of Rheumatism. Besides having many uses and folklore claims, herbal medicines are to be thoroughly investigated for their toxicity also. Therefore this work is being carried out to examine the toxicity of the drug and established dose is safe to use in the clinical stage. The current research studied the acute and chronic toxicity of Polygonum glabrum root extract in rats. It is proved that there was no change in any parameter tested both in acute and chronic toxicity, which means the extract is safe and non-toxic at the dose of 2g/kg also.

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Naphthoquinones and derivatives for chemotherapy: perspectives and limitations of their anti-trypanosomatids activities

Current Pharmaceutical Design ◽

10.2174/1381612826666201109111802 ◽

2020 ◽

Vol 26 ◽

Author(s):

Luíza Dantas-Pereira ◽

Edézio F. Cunha-Junior ◽

Valter V. Andrade-Neto ◽

John F. Bower ◽

Guilherme A. M. Jardim ◽

...

Keyword(s):

Large Scale ◽

Neglected Tropical Diseases ◽

Folk Medicine ◽

Leishmania Species ◽

Parasitic Infections ◽

Mechanistic Analysis ◽

Leishmania Spp ◽

Nanomolar Range

: Chagas disease, Sleeping sickness and Leishmaniasis, caused by trypanosomatids Trypanosoma cruzi, Trypanosoma brucei and Leishmania spp., respectively, are considered neglected tropical diseases, and they especially affect impoverished populations in the developing world. The available chemotherapies are very limited and a search for alternatives is still necessary. In folk medicine, natural naphthoquinones have been employed for the treatment of a great variety of illnesses, including parasitic infections. This review is focused on the anti-trypanosomatid activity and mechanistic analysis of naphthoquinones and derivatives. Among all the series of derivatives tested in vitro, naphthoquinone-derived 1,2,3-triazoles were very active on T. cruzi infective forms in blood bank conditions, as well as in amastigotes of Leishmania spp. naphthoquinones containing a CF3 on a phenyl amine ring inhibited T. brucei proliferation in the nanomolar range, and naphthopterocarpanquinones stood out for their activity on a range of Leishmania species. Some of these compounds showed a promising selectivity index (SI) (30 to 1900), supporting further analysis in animal models. Indeed, high toxicity to the host and inactivation by blood components are crucial obstacles to be overcome to use naphthoquinones and/or their derivatives for chemotherapy. Multidisciplinary initiatives embracing medicinal chemistry, bioinformatics, biochemistry, and molecular and cellular biology need to be encouraged to allow the optimization of these compounds. Large scale automated tests are pivotal for the efficiency of the screening step, and subsequent evaluation of both the mechanism of action in vitro and pharmacokinetics in vivo are essential for the development of a novel, specific and safe derivative, minimizing adverse effects.

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Implantable neural interfaces

10.1093/oso/9780199674923.003.0050 ◽

2018 ◽

Author(s):

Stefano Vassanelli

Keyword(s):

Brain Function ◽

Large Scale ◽

Ionic Channels ◽

Patch Clamp Technique ◽

Advantages And Disadvantages ◽

Optogenetic Stimulation ◽

Physical Interfaces ◽

The Brain

Establishing direct communication with the brain through physical interfaces is a fundamental strategy to investigate brain function. Starting with the patch-clamp technique in the seventies, neuroscience has moved from detailed characterization of ionic channels to the analysis of single neurons and, more recently, microcircuits in brain neuronal networks. Development of new biohybrid probes with electrodes for recording and stimulating neurons in the living animal is a natural consequence of this trend. The recent introduction of optogenetic stimulation and advanced high-resolution large-scale electrical recording approaches demonstrates this need. Brain implants for real-time neurophysiology are also opening new avenues for neuroprosthetics to restore brain function after injury or in neurological disorders. This chapter provides an overview on existing and emergent neurophysiology technologies with particular focus on those intended to interface neuronal microcircuits in vivo. Chemical, electrical, and optogenetic-based interfaces are presented, with an analysis of advantages and disadvantages of the different technical approaches.

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What Has Direct Cortical and Subcortical Electrostimulation Taught Us about Neurolinguistics?

The Oxford Handbook of Neurolinguistics ◽

10.1093/oxfordhb/9780190672027.013.8 ◽

2019 ◽

pp. 185-211

Author(s):

Hugues Duffau

Keyword(s):

White Matter ◽

Large Scale ◽

Functional Neuroimaging ◽

Great Accuracy ◽

Subcortical White Matter ◽

Cerebral Lesions ◽

Physiological Basis ◽

Postoperative Mri ◽

The Brain

Investigating the neural and physiological basis of language is one of the most important challenges in neurosciences. Direct electrical stimulation (DES), usually performed in awake patients during surgery for cerebral lesions, is a reliable tool for detecting both cortical and subcortical (white matter and deep grey nuclei) regions crucial for cognitive functions, especially language. DES transiently interacts locally with a small cortical or axonal site, but also nonlocally, as the focal perturbation will disrupt the entire subnetwork sustaining a given function. Thus, in contrast to functional neuroimaging, DES represents a unique opportunity to identify with great accuracy and reproducibility, in vivo in humans, the structures that are actually indispensable to the function, by inducing a transient virtual lesion based on the inhibition of a subcircuit lasting a few seconds. Currently, this is the sole technique that is able to directly investigate the functional role of white matter tracts in humans. Thus, combining transient disturbances elicited by DES with the anatomical data provided by pre- and postoperative MRI enables to achieve reliable anatomo-functional correlations, supporting a network organization of the brain, and leading to the reappraisal of models of language representation. Finally, combining serial peri-operative functional neuroimaging and online intraoperative DES allows the study of mechanisms underlying neuroplasticity. This chapter critically reviews the basic principles of DES, its advantages and limitations, and what DES can reveal about the neural foundations of language, that is, the large-scale distribution of language areas in the brain, their connectivity, and their ability to reorganize.

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Optimizing Manufacturing and Osseointegration of Ti6Al4V Implants through Precision Casting and Calcium and Phosphorus Ion Implantation? In Vivo Results of a Large-Scale Animal Trial

Materials ◽

10.3390/ma13071670 ◽

2020 ◽

Vol 13 (7) ◽

pp. 1670 ◽

Cited By ~ 2

Author(s):

Wölfle-Roos JV ◽

Katmer Amet B ◽

Fiedler J ◽

Michels H ◽

Kappelt G ◽

...

Keyword(s):

Ion Implantation ◽

Large Scale ◽

In Vivo Studies ◽

Control Group ◽

Implant Surface ◽

Precision Casting ◽

Pull Out ◽

Significant Difference ◽

Calcium And Phosphorus

Background: Uncemented implants are still associated with several major challenges, especially with regard to their manufacturing and their osseointegration. In this study, a novel manufacturing technique—an optimized form of precision casting—and a novel surface modification to promote osseointegration—calcium and phosphorus ion implantation into the implant surface—were tested in vivo. Methods: Cylindrical Ti6Al4V implants were inserted bilaterally into the tibia of 110 rats. We compared two generations of cast Ti6Al4V implants (CAST 1st GEN, n = 22, and CAST 2nd GEN, n = 22) as well as cast 2nd GEN Ti6Al4V implants with calcium (CAST + CA, n = 22) and phosphorus (CAST + P, n = 22) ion implantation to standard machined Ti6Al4V implants (control, n = 22). After 4 and 12 weeks, maximal pull-out force and bone-to-implant contact rate (BIC) were measured and compared between all five groups. Results: There was no significant difference between all five groups after 4 weeks or 12 weeks with regard to pull-out force (p > 0.05, Kruskal Wallis test). Histomorphometric analysis showed no significant difference of BIC after 4 weeks (p > 0.05, Kruskal–Wallis test), whereas there was a trend towards a higher BIC in the CAST + P group (54.8% ± 15.2%), especially compared to the control group (38.6% ± 12.8%) after 12 weeks (p = 0.053, Kruskal–Wallis test). Conclusion: In this study, we found no indication of inferiority of Ti6Al4V implants cast with the optimized centrifugal precision casting technique of the second generation compared to standard Ti6Al4V implants. As the employed manufacturing process holds considerable economic potential, mainly due to a significantly decreased material demand per implant by casting near net-shape instead of milling away most of the starting ingot, its application in manufacturing uncemented implants seems promising. However, no significant advantages of calcium or phosphorus ion implantation could be observed in this study. Due to the promising results of ion implantation in previous in vitro and in vivo studies, further in vivo studies with different ion implantation conditions should be considered.

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