Impact of Nitrogen Fertilization on Phytophthora cinnamomi Root-related Damage in Juglans regia Saplings

Excessive nitrogen (N) use in agriculture has been associated with increased severity of the damage caused by Phytophthora species. In this study, we investigated the impact in vitro and in vivo of N about Phytophthora cinnamomi. The preliminary in vitro assay showed the effect of different N sources on the mycelial growth of P. cinnamomi. This assay indicated that ammonium nitrate (NH4NO3) and ammonium sulfate [(NH4)2SO4] allowed for greater control of P. cinnamomi mycelia in comparison with calcium nitrate [Ca(NO3)2] and potassium nitrate (KNO3) when used with 1000 ppm N. The in vivo assay showed the severity of P. cinnamomi in 5-month-old Juglans regia saplings grown under greenhouse conditions. We selected NH4NH3 as the source for N for the greenhouse assay, considering the inhibitory effect on the ingrowth of P. cinnamomi and the intensive use of this fertilizer in agriculture. Walnut saplings were fertilized with 0, 35, 70, 140, 210, and 1050 ppm N and were inoculated with zoospores of P. cinnamomi 45 d after the application of nitrogen treatment (DAA). They were harvested at 90 DAA. We found that a 70-ppm N fertilization reduced the development of P. cinnamomi, resulting in lower root and canopy damage indices (DIs) than the unfertilized inoculated treatments and fertilized treatments greater than 140 ppm. The results of the in vitro and in vivo assay agree that increased N concentrations were associated with reduced mycelium growth of P. cinnamomi, providing further evidence that N fertilization can mitigate this disease. Greater root and canopy damage was observed in saplings fertilized with 1050 ppm N, regardless of whether they were inoculated with P. cinnamomi, as a result of N phytotoxicity (verified through foliar analysis). In contrast, inoculated and unfertilized saplings (N0) also showed high root and canopy DIs associated either with the inoculation with P. cinnamomi or the no fertilization treatment. We postulate that 70 ppm N is the best fertilization rate for J. regia saplings because the positive effects of N on growth are maximized and the damage caused by P. cinnamomi is mitigated.

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Effect of Massoia (Massoia aromatica Becc.) Bark on the Phagocytic Activity of Wistar Rat Macrophages

10.20944/preprints201804.0028.v1 ◽

2018 ◽

Author(s):

Triana Hertiani ◽

Agustinus Yuswanto ◽

Sylvia Utami Tunjung Pratiwi ◽

Harlyanti Mashar

Keyword(s):

Phagocytic Activity ◽

Wistar Rats ◽

In Vitro Assay ◽

Phagocytic Index ◽

Dependent Manner ◽

Vitro Assay ◽

Macrophage Cells ◽

In Vivo Assay

Massoia (Massoia aromatica Becc., Lauraceae) bark has been widely used as a component of traditional Indonesian medicine. The indigenous people boil or steam the bark for traditional applications. Our preliminary research revealed the potency of Massoia essential oil and its major compound, C-10 Massoialactone as potential immunomodulator in vitro. However, no scientific evidence regarding its in vivo effects is available. Therefore, this study evaluated the potential immunomodulatory effects of Massoia bark infusion on the nonspecific immune response (phagocytosis) of Wistar rats. The aqueous extract of Massoia bark was obtained by boiling pulverized bark in water, and the C-10 massoialactone content of the extract was determined through Thin Layer Chromatography (TLC) densitometry. For the in vitro assay, macrophages were treated with the freeze-dried infusion at the concentrations of 2.5, 5, 10, 20, or 40 μg/mL media. For the in vivo assay, 2-month-old male Wistar rats were divided into 5 groups. The baseline group received distilled water at the dose of 1 mL/100 g BW with the immunostimulant herbal product “X” administered as the positive control at the dose of 0.54 mL/rat. The treatment groups received the infusion at a dose of 100, 300, or 500 mg/100 g BW. Treatments were given orally every day for 14 days. The ability of macrophage cells to phagocyte latex was determined as phagocytic index (PI) and was observed under microscopy with 300 macrophages. The in vitro study revealed that the phagocytic activity of the infusion-treated macrophages significantly increased in comparison with that of the control macrophages in a concentration-dependent manner. Among all treatment concentrations, the concentration of 40 μg/ml provided the highest activity with a PI value of 70.51% ± 1.11%. The results of the in vivo assay confirmed those of the in vitro assay. The results of the present study indicate that Massoia bark can increase the phagocytic activity of rat macrophage cells. Its potential as a naturally derived immunomodulatory agent requires further study.

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Alternative Splicing of FN (Fibronectin) Regulates the Composition of the Arterial Wall Under Low Flow

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.120.314013 ◽

2020 ◽

Author(s):

Patrick A. Murphy ◽

Noor Jailkhani ◽

Sarah-Anne Nicholas ◽

Amanda M. Del Rosario ◽

Jeremy L. Balsbaugh ◽

...

Keyword(s):

Extracellular Matrix ◽

Alternative Splicing ◽

Low Flow ◽

Matrix Composition ◽

Splice Isoforms ◽

Vitro Assay ◽

Disturbed Flow ◽

The Impact

Objective: Exposure of the arterial endothelium to low and disturbed flow is a risk factor for the erosion and rupture of atherosclerotic plaques and aneurysms. Circulating and locally produced proteins are known to contribute to an altered composition of the extracellular matrix at the site of lesions, and to contribute to inflammatory processes within the lesions. We have previously shown that alternative splicing of FN (fibronectin) protects against flow-induced hemorrhage. However, the impact of alternative splicing of FN on extracellular matrix composition remains unknown. Approach and Results: Here, we perform quantitative proteomic analysis of the matrisome of murine carotid arteries in mice deficient in the production of FN splice isoforms containing alternative exons EIIIA and EIIIB (FN-EIIIAB null) after exposure to low and disturbed flow in vivo. We also examine serum-derived and endothelial-cell contributions to the matrisome in a simplified in vitro system. We found flow-induced differences in the carotid artery matrisome that were impaired in FN-EIIIAB null mice. One of the most interesting differences was reduced recruitment of FBLN1 (fibulin-1), abundant in blood and not locally produced in the intima. This defect was validated in our in vitro assay, where FBLN1 recruitment from serum was impaired by the absence of these alternatively spliced segments. Conclusions: Our results reveal the extent of the dynamic alterations in the matrisome in the acute response to low and disturbed flow and show how changes in the splicing of FN, a common response in vascular inflammation and remodeling, can affect matrix composition.

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Efficacy of Ocimum gratissimum essential oil against the monogenean Cichlidogyrus tilapiae gill parasite of Nile tilapia

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/1678-4162-9667 ◽

2018 ◽

Vol 70 (2) ◽

pp. 497-504 ◽

Cited By ~ 6

Author(s):

J.O. Meneses ◽

M.V.S. do Couto ◽

N.C. Sousa ◽

F. dos S. Cunha ◽

H.A. Abe ◽

...

Keyword(s):

Essential Oil ◽

Nile Tilapia ◽

Anthelmintic Activity ◽

In Vitro Assay ◽

Vitro Assay ◽

Ocimum Gratissimum ◽

In Vivo Assay

ABSTRACT The phythotherapy is an alternative to use of chemotherapeutical agents against monogenean infection. This study evaluated the anthelmintic activity of essential oil Ocimum gratissimum against monogenean Cichlidogyrus tilapiae as well as its acute toxicity in tilapia juveniles. The mean lethal concentration (LC50) and different concentrations of the essential oil, both in vitro and in vivo assays (short and long-term baths) were assessed. The LC50 was 40.70mg.L-1 and in the in vitro assay this concentration showed 80% efficacy at the last two hours and in the in vivo assay 65.87% efficacy in long-term bath. However, it provoked morphological alterations on the gills such as hyperplasia and edema. The parasites exposure at the highest concentration (320mg.L-1) showed 100% mortality after 2h exposure in the in vitro assay, whereas in the in vivo assay, short-term baths of 5min for 3 consecutive days showed an efficacy of 87.71% without gills damage. These results demonstrate the anthelminthic activity of essential oil O. gratissimum and the safety concentration to use in Nile tilapia.

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Effects of leptin replacement on hypothalamic-pituitary growth hormone axis function and circulating ghrelin levels in ob/ob mice

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00258.2006 ◽

2007 ◽

Vol 292 (3) ◽

pp. E891-E899 ◽

Cited By ~ 47

Author(s):

Raul M. Luque ◽

Zhi H. Huang ◽

Bhumik Shah ◽

Theodore Mazzone ◽

Rhonda D. Kineman

Keyword(s):

Growth Hormone ◽

Food Intake ◽

Statistical Significance ◽

Mrna Levels ◽

Ghrelin Receptor ◽

Positive Effects ◽

In Vivo Effects ◽

The Impact

Leptin-deficient obese mice ( ob/ob) have decreased circulating growth hormone (GH) and pituitary GH and ghrelin receptor (GHS-R) mRNA levels, whereas hypothalamic GH-releasing hormone (GHRH) and somatostatin (SST) expression do not differ from lean controls. Given the fact that GH is suppressed in diet-induced obesity (a state of hyperleptinemia), it remains to be determined whether the absence of leptin contributes to changes in the GH axis of ob/ob mice. Therefore, to study the impact of leptin replacement on the hypothalamic-pituitary GH axis of ob/ob mice, leptin was infused for 7 days (sc), resulting in circulating leptin levels that were similar to wild-type controls (∼1 ng/ml). Leptin treatment reduced food intake, body weight, and circulating insulin while elevating circulating n-octanoyl ghrelin concentrations. Leptin treatment did not alter hypothalamic GHRH, SST, or GHS-R mRNA levels compared with vehicle-treated controls. However, leptin significantly increased pituitary GH and GHRH-R expression and tended to enhance circulating GH levels, but this latter effect did not reach statistical significance. In vitro, leptin (1 ng/ml, 24 h) did not affect pituitary GH, GHRH-R, or GHS-R mRNA but did enhance GH release. The in vivo effects of leptin on circulating hormone and pituitary mRNA levels were not replicated by pair feeding ob/ob mice to match the food intake of leptin-treated mice. However, leptin did prevent the fall in hypothalamic GHRH mRNA and circulating IGF-I levels observed in pair-fed mice. These results demonstrate that leptin replacement has positive effects on multiple levels of GH axis function in ob/ob mice.

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Control of Meloidogyne incognita in tomato plants with highly diluted solutions of Thuya occidentalis and their effects on plant growth and defense metabolism

Semina Ciências Agrárias ◽

10.5433/1679-0359.2017v38n4p2187 ◽

2017 ◽

Vol 38 (4) ◽

pp. 2187

Author(s):

Thaísa Muriel Mioranza ◽

José Renato Stangarlin ◽

Mônica Anghinoni Müller ◽

Sidiane Coltro-Roncato ◽

Cristiane Claudia Meinerz ◽

...

Keyword(s):

Meloidogyne Incognita ◽

Defense Responses ◽

Control Treatment ◽

Root Knot Nematode ◽

Vitro Assay ◽

Vitro Experiment ◽

In Vitro Experiment ◽

In Vivo Assay

This work aimed to control the root-knot nematode Meloidogyne incognita in tomato Solanum lycopersicum L plants with high-diluted solutions of Thuya occidentalis, and to study its effects on growth and plant defense responses. The in vivo experiment was carried out over two years (2013 and 2014) at a climatized greenhouse, whilst the in vitro experiment was carried out in the laboratory. Eight treatments were used (6, 12, 24, 50, 100, 200 and 400CH (Hahnemannian centesimal) of T. occidentalis, with water as control treatment). For the in vivo assay, in 2013 plants were inoculated with about 4850 eggs and second-stage juveniles (J2), while in 2014 they were inoculated with 5050 eggs and J2. The treatments were applied once a week, as 0.1% aqueous solutions onto the plant shoots, for 50 and 40 days respectively. For the in vitro experiment, the nematodes were directly exposed to the same 0.1% treatments. The treatments did not show nematostatic or nematicide effects in the in vitro assay, and had no influence on the hatching of the eggs. For the in vivo assay in 2013, T. occidentalis 100CH decreased the number of J2 present in the roots, whilst the dynamization of 200CH stimulated root development and increased the weight of the fruits of the first cluster. In 2014, 100CH decreased numbers of J2 in the soil. Some dynamization increased the plant’s defense enzyme activity, such as peroxidase (24, 50, 200 and 400CH), polyphenoloxidase (200CH), and phenylalanine ammonia-lyase (24 and 50CH). In this study, T. occidentalis 100CH showed potential for the control of M. incognita, whilst 24 and 200CH influenced the growth of plants.

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Evaluation of pro-angiogenic properties of an inorganic silica gel fibre fleece

Journal of Wound Care ◽

10.12968/jowc.2021.30.9.729 ◽

2021 ◽

Vol 30 (9) ◽

pp. 729-736

Author(s):

Vera Grotheer ◽

Joachim Windolf ◽

Christoph V Suschek

Keyword(s):

Inorganic Materials ◽

Wound Dressings ◽

Orthosilicic Acid ◽

Angiogenic Potential ◽

Positive Effects ◽

Wound Model ◽

Angiogenesis Assay ◽

The Impact

Hard-to-heal wounds represent an increasing health and economic burden on society. At present, therapy options for hard-to-heal wounds are often unsatisfactory, and the development of more effective wound treatments is urgently needed. We have shown that orthosilicic acid-releasing silica fibre fleece (SIFIB), via its pronounced anti-inflammatory properties, exhibited a significantly enhanced effect on wound closure kinetics in a porcine wound model in vivo. In this present study, we have examined in vitro the impact of the pro-angiogenic potential of SIFIB. Using an in vitro angiogenesis assay we describe for the first time how an inorganic biodegradable silica-based material significantly improved endothelial microvessel-like structure formation. We further demonstrate that the molecular mechanism of this pro-angiogenic activity of SIFIB is based on a significantly increased and tumour necrosis factor (TNF)α-dependent VEGF protein expression. In conclusion, due to its positive effects on angiogenesis, our results further indicate that decomposition products of silica-based biodegradable inorganic materials might represent very relevant therapeutic components of modern wound dressings for the treatment of hard-to-heal wounds.

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Effects of Probiotics on Growth, Survival, Water Quality and Disease Resistance of Red Hybrid Tilapia (Oreochromis spp.) Fingerlings in a Biofloc System

Animals ◽

10.3390/ani11123514 ◽

2021 ◽

Vol 11 (12) ◽

pp. 3514

Author(s):

Aimi Zabidi ◽

Fatimah Md Yusoff ◽

S. M. Nurul Amin ◽

Nur Jasmin Mohd Yaminudin ◽

Puvaneswari Puvanasundram ◽

...

Keyword(s):

Disease Resistance ◽

Growth Performance ◽

Nitrogen Concentration ◽

Feed Conversion ◽

Vitro Assay ◽

Beneficial Effects ◽

Positive Effects ◽

Commercial Probiotics

Biofloc technology has shown positive effects in aquaculture, especially on the growth performance of cultured animals. The aims of this study were to evaluate the effects of adding different probiotic strains in a biofloc system on the growth performance and disease resistance of red hybrid tilapia (Oreochromis spp.). Three different probiotics (Lysinibacillus fusiformis SPS11, Bacillus amyloliquefaciens L9, and Enterococcus hirae LAB3), commercial probiotics (MG1) and a mixed probiotics (MP) combining all three strains were used in this study. The in vitro assay results showed that the mixed probiotic (MP) was able to inhibit the growth of Streptococcus agalactiae and Streptococcus iniae significantly compared to the single and commercial probiotic. The efficacy of MP was further tested in in vivo tilapia culture challenged with S. agalactiae. The best specific growth rate (3.73 ± 0.23% day−1) and feed conversion ratio (0.76 ± 0.04) were recorded in the group of biofloc with addition of MP. After being challenged with S. agalactiae, the group of biofloc with MP had significantly higher survival (83 ± 1.43%) compared to the other groups. Furthermore, the nitrogen concentration (NO2-N and NH4-N) was significantly lower in all the biofloc groups compared to the control. Hence, the addition of probiotics was able to provide beneficial effects to red hybrid tilapia culture in the biofloc system.

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Further Considerations Towards an Effective and Efficient Oncology Drug Discovery DMPK Strategy

Current Drug Metabolism ◽

10.2174/1389200221666200312104837 ◽

2020 ◽

Vol 21 (2) ◽

pp. 145-162 ◽

Cited By ~ 1

Author(s):

Beth Williamson ◽

Nicola Colclough ◽

Adrian John Fretland ◽

Barry Christopher Jones ◽

Rhys Dafydd Owen Jones ◽

...

Keyword(s):

Drug Discovery ◽

Drug Exposure ◽

Phase Iii ◽

Human Pharmacokinetics ◽

Vitro Assay ◽

Pharmacodynamic Modelling ◽

Oncology Drug ◽

The Impact

Background: DMPK data and knowledge are critical in maximising the probability of developing successful drugs via the application of in silico, in vitro and in vivo approaches in drug discovery. Methods: The evaluation, optimisation and prediction of human pharmacokinetics is now a mainstay within drug discovery. These elements are at the heart of the ‘right tissue’ component of AstraZeneca’s ‘5Rs framework’ which, since its adoption, has resulted in increased success of Phase III clinical trials. With the plethora of DMPK related assays and models available, there is a need to continually refine and improve the effectiveness and efficiency of approaches best to facilitate the progression of quality compounds for human clinical testing. Results: This article builds on previously published strategies from our laboratories, highlighting recent discoveries and successes, that brings our AstraZeneca Oncology DMPK strategy up to date. We review the core aspects of DMPK in Oncology drug discovery and highlight data recently generated in our laboratories that have influenced our screening cascade and experimental design. We present data and our experiences of employing cassette animal PK, as well as re-evaluating in vitro assay design for metabolic stability assessments and expanding our use of freshly excised animal and human tissue to best inform first time in human dosing and dose escalation studies. Conclusion: Application of our updated drug-drug interaction and central nervous system drug exposure strategies are exemplified, as is the impact of physiologically based pharmacokinetic and pharmacokinetic-pharmacodynamic modelling for human predictions.

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Physiologically-Based Pharmacokinetic (PBPK) Modelling of Transporter Mediated Drug Absorption, Clearance and Drug-Drug Interactions

Current Drug Metabolism ◽

10.2174/1389200221999210101233340 ◽

2021 ◽

Vol 21 ◽

Author(s):

Kunal S. Taskar ◽

Isobel Harada ◽

Ravindra V. Alluri

Keyword(s):

Drug Interactions ◽

Intestinal Absorption ◽

Drug Disposition ◽

Pbpk Modelling ◽

Vitro Assay ◽

Pbpk Models ◽

In Vitro Systems ◽

The Impact

Abstarct: Membrane transporters play an important role in intestinal absorption, distribution and clearance of drugs. Additionally transporters along with enzymes regulate tissue exposures (e.g. liver, kidney and brain), which are important for safety and efficacy considerations. Early identification of transporters involved guides generation of in vitro and in vivo data needed to gain mechanistic understanding on the role of transporters in organ clearance, tissue exposures and enables development of physiological-based pharmacokinetic (PBPK) models. A lot of progress has been made in developing several in vitro assay systems and mechanistic in silico models to determine kinetic parameters for transporters, which are incorporated into PBPK models. Although, intrinsic clearance and inhibition data from in vitro systems generally tend to underpredict in vivo clearance and magnitude of drug-drug interactions (DDIs), empirical scaling factors derived from a sizable dataset are often used to offset underpredictions. PBPK models are increasing used to predict the impact of transporters on intestinal absorption, clearance, victim and perpetrator DDIs prior to first in human clinical trials. The models are often refined when clinical data is available and are used to predict pharmacokinetics in untested scenarios such as the impact of polymorphisms, ontogeny, ethnicity, disease states and DDIs with other perpetrator drugs. The aim of this review is to provide an overview of (i) regulatory requirements around transporters, (ii) in vitro systems and their limitations in predicting transporter mediated drug disposition and DDIs, (iii) PBPK modelling tactics and case studies used for internal decision making and/or for regulatory submissions.

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