Monocytes expressing activin A and CCR2 exacerbate chronic testicular inflammation by promoting immune cell infiltration.

STUDY QUESTION: Does the chemokine/chemokine receptor axis, involved in immune cell trafficking, contribute to the pathology of testicular inflammation and how does activin A modulate this network? SUMMARY ANSWER: Testicular chemokines and their receptors (especially those essential for trafficking of monocytes) are elevated in orchitis, and activin A modulates the expression of the chemokine/chemokine receptor network to promote monocyte/macrophage and T cell infiltration into the testes, causing extensive tissue damage. WHAT IS KNOWN ALREADY: The levels of CC motif chemokine receptor (CCR)2 and its ligand CC motif chemokine ligand (CCL)2 are increased in experimental autoimmune orchitis (EAO) compared with healthy testes, and mice deficient in CCR2 are protected from EAO-induced tissue damage. Activin A induces CCR2 expression in macrophages, promoting their migration. Moreover, there is a positive correlation between testicular activin A concentration and the severity of autoimmune orchitis. Inhibition of activin A activity by overexpression of follistatin (FST) reduces EAO-induced testicular damage. STUDY DESIGN, SIZE, DURATION: EAO was induced in 10-12-week-old male C57BL/6J (wild-type; WT) and B6.129P2-Ccr2tm1Mae/tm1Mae (Ccr2-/-) mice (n = 6). Adjuvant (n = 6) and untreated (n = 6) age-matched control mice were also included. Testes were collected at 50 days after the first immunization with testicular homogenate in complete Freund's adjuvant. In another experimental setup, WT mice were injected with a non-replicative recombinant adeno-associated viral vector carrying a FST315-expressing gene cassette (rAAV-FST315; n = 7-9) or an empty control vector (n = 5) 30 days prior to EAO induction. Appropriate adjuvant (n = 4-5) and untreated (n = 4-6) controls were also examined. Furthermore, human testicular biopsies exhibiting focal leukocytic infiltration and impaired spermatogenesis (n = 17) were investigated. Biopsies showing intact spermatogenesis were included as controls (n = 9). Bone-marrow-derived macrophages (BMDMs) generated from WT mice were treated with activin A (50 ng/ml) for 6 days. Activin-A-treated or untreated BMDMs were then co-cultured with purified mouse splenic T cells for two days to assess chemokine and cytokine production. PARTICIPANTS/MATERIALS, SETTING, METHODS: Quantitative real-time PCR (qRT-PCR) was used to analyze the expression of chemokines in total testicular RNA collected from mice. Immunofluorescence staining was used to detect activin A, F4/80, and CD3 expression in mouse testes. The expression of chemokine/chemokine-receptor-encoding genes was examined in human testicular biopsies by qRT-PCR. Correlations between chemokine expression levels and either the immune cell infiltration density or the mean spermatogenesis score were analyzed. Immunofluorescence staining was used to evaluate the expression of CD68 and CCR2 in human testicular biopsies. RNA isolated from murine BMDMs was used to characterize these cells in terms of their chemokine/chemokine receptor expression levels. Conditioned media from co-cultures of BMDMs and T cells were collected to determine chemokine levels and the production of pro-inflammatory cytokines tumor necrosis factor (TNF) and interferon (IFN)-γ by T cells. MAIN RESULTS AND THE ROLE OF CHANCE: Induction of EAO in the testes of WT mice increased the expression of chemokine receptors such as Ccr1 (P < 0.001), Ccr2 (P < 0.0001), Ccr3 (P < 0.0001), Ccr5 (P < 0.0001), CXC motif chemokine receptor (Cxcr)3 (P < 0.01), and CX3C motif chemokine receptor (Cx3cr)1 (P < 0.001), as well as that of most of their ligands. Ccr2 deficiency reversed some of the changes associated with EAO by reducing the expression of Ccr1 (P < 0.0001), Ccr3 (P < 0.0001), Ccr5 (P < 0.01), Cxcr3 (P < 0.001), and Cx3cr1 (P < 0.0001). Importantly, the biopsies showing impaired spermatogenesis and concomitant focal leukocytic infiltration exhibited higher expression of CCL2 (P < 0.01), CCR1 (P < 0.05), CCR2 (P < 0.001), and CCR5 (P < 0.001) than control biopsies with no signs of inflammation and intact spermatogenesis. The gene expression of CCR2 and its ligand CCL2 correlated positively with the immune cell infiltration density (P < 0.05) and negatively with the mean spermatogenesis score (P < 0.001). Moreover, CD68+ macrophages expressing CCR2 were present in human testes with leukocytic infiltration with evidence of tubular damage. Treatment of BMDMs, as surrogates for testicular macrophages, with activin A increased their expression of Ccr1, Ccr2, and Ccr5 while reducing their expression of Ccl2, Ccl3, Ccl4, Ccl6, Ccl7 Ccl8, and Ccl12. These findings were validated in vivo, by showing that inhibiting activin A activity by overexpressing FST in EAO mice decreased the expression of Ccr2 (P < 0.05) and Ccr5 (P < 0.001) in the testes. Interestingly, co-culturing activin-A-treated BMDMs and T cells reduced the levels of CCL2 (P < 0.05), CCL3/4 (P < 0.01), and CCL12 (P < 0.05) in the medium and attenuated the production of TNF (P < 0.05) by T cells. The majority of cells secreting activin A in EAO testes were identified as macrophages. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: BMDMs were used as surrogates for testicular macrophages. Hence, results obtained from the in vitro experiments might not be fully representative of the situation in the testes in vivo. Moreover, since total RNA was extracted from the testicular tissue to examine chemokine expression, the contributions of individual cell types as producers of specific chemokines may have been overlooked. WIDER IMPLICATIONS OF THE FINDINGS: Our data indicate that macrophages are implicated in the development and progression of testicular inflammation by expressing CCR2 and activin A, which ultimately remodel the chemokine/chemokine receptor network and recruit other immune cells to the site of inflammation. Consequently, inhibition of CCR2 or activin A could serve as a potential therapeutic strategy for reducing testicular inflammation. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the International Research Training Group in 'Molecular pathogenesis on male reproductive disorders', a collaboration between Justus Liebig University (Giessen) and Monash University (Melbourne) (GRK1871/1-2) funded by the Deutsche Forschungsgemeinschaft and Monash University, a National Health and Medical Research Council of Australia Ideas Grant (1184867), and the Victorian Government's Operational Infrastructure Support Programme. The authors declare no competing financial interests.

Activin A and CCR2 regulate macrophage function in testicular fibrosis caused by experimental autoimmune orchitis.

Experimental autoimmune-orchitis (EAO), a rodent model of chronic testicular inflammation and fibrosis, replicates pathogenic changes seen in some cases of human spermatogenic disturbances. During EAO, increased levels of pro-inflammatory and pro-fibrotic mediators such as TNF, CCL2, and activin A are accompanied by infiltration of leukocytes into the testicular parenchyma. Activin A levels correlate with EAO severity, while elevated CCL2 acting through its receptor CCR2 mediates leukocyte trafficking and recruits macrophages. CCR2 + CXCR4 + macrophages producing extracellular matrix proteins contribute widely to fibrogenesis. Furthermore, testicular macrophages (TMs) play a critical role in organ homeostasis. Therefore, we aimed to investigate the role of the activin A/CCL2-CCR2/macrophage axis in the development of testicular fibrosis. Following EAO induction, we observed lower levels of organ damage, collagen deposition, and leukocyte infiltration (including fibronectin+, collagen I+ and CXCR4+ TMs) in Ccr2-/- mice than in WT mice. Furthermore, levels of Il-10, Ccl2, and the activin A subunit Inhba mRNAs were lower in Ccr2-/- EAO testes. Notably, fibronectin+ TMs were also present in biopsies from patients with impaired spermatogenesis and fibrotic alterations. Overexpression of the activin A antagonist follistatin reduced tissue damage and collagen I+ TM accumulation in WT EAO testes, while treating macrophages with activin A in vitro increased the expression of Ccr2, Fn1, Cxcr4, and Mmp2 and enhanced migration along a CCL2 gradient; these effects were abolished by follistatin. Taken together, our data indicate that CCR2 and activin A promote fibrosis during testicular inflammation by regulating macrophage function. Inhibition of CCR2 or activin A protects against damage progression, offering a promising avenue for therapeutic intervention.

Potential Adverse Effects of Resveratrol: A Literature Review.

Due to its health benefits, resveratrol (RE) is one of the most researched natural polyphenols. Resveratrol's health benefits were first highlighted in the early 1990s in the French paradox study, which opened extensive research activity into this compound. Ever since, several pharmacological activities including antioxidant, anti-aging, anti-inflammatory, anti-cancerous, anti-diabetic, cardioprotective, and neuroprotective properties, were attributed to RE. However, results from the available human clinical trials were controversial concerning the protective effects of RE against diseases and their sequelae. The reason for these conflicting findings is varied but differences in the characteristics of the enrolled patients, RE doses used, and duration of RE supplementation were proposed, at least in part, as possible causes. In particular, the optimal RE dosage capable of maximizing its health benefits without raising toxicity issues remains an area of extensive research. In this context, while there is a consistent body of literature on the protective effects of RE against diseases, there are relatively few reports investigating its possible toxicity. Indeed, toxicity and adverse effects were reported following consumption of RE; therefore, extensive future studies on the long-term effects, as well as the in vivo adverse effects, of RE supplementation in humans are needed. Furthermore, data on the interactions of RE when combined with other therapies are still lacking, as well as results related to its absorption and bioavailability in the human body. In this review, we collect and summarize the available literature about RE toxicity and side effects. In this process, we analyze in vitro and in vivo studies that have addressed this stilbenoid. These studies suggest that RE still has an unexplored side. Finally, we discuss the new delivery methods that are being employed to overcome the low bioavailability of RE.

Therapeutic benefits of Indole-3-Carbinol in adjuvant-induced arthritis and its protective effect against methotrexate induced-hepatic toxicity.

BACKGROUND: Rheumatoid arthritis (RA) being an incapacitating disease requires early effective intervention. Considering Methotrexate (MTX)- the first line of treatment for RA- intoxicates the liver; therefore, alternative therapies with similar efficacy yet lower cytotoxicity are desired. Indole-3-Carbinol (I3C) which is found in cruciferous vegetables was examined for its possible therapeutic potentials in comparison with MTX by investigating its anti-inflammatory, anti-arthritic, anti-oxidant, and hepatoprotective potentials in adjuvant-induced arthritis (AIA) rat model. METHODS: Arthritis was induced in Sprague Dawley rats by injection of Complete Freund's Adjuvant (CFA). Arthritic rats were treated with I3C and/or MTX. To examine the anti-inflammatory and anti-arthritic effect, the following parameters were assessed: body weight, macroscopic scoring of the hind paw, the level of the pivotal cytokines (TNF-α, IL-6) heavily involved in the pathogenesis, spleen index, and erythrocyte sedimentation rate. At a histological level, the tibiotarsal joint was stained with several specific stains. To assess the hepatoprotective and anti-oxidant effects, several oxidative stress parameters were monitored, and the liver histology was examined. RESULTS: Both I3C and MTX attenuated the inflammation that was aggravated by arthritis by downregulating the inflammatory markers and mediators and alleviating the histopathological changes affecting the tibiotarsal joint. I3C attenuated the liver impairment that was initiated by arthritis and MTX treatment. It did so by downregulating the pro-oxidants and up-regulating the anti-oxidant defenses and by reducing the pathological changes affecting the liver. CONCLUSION: Our results suggest that I3C is as potent as MTX as an anti-inflammatory and anti-arthritic agent. In addition, I3C does so while protecting the liver from damage as opposed to MTX.

Mechanism of Inflammatory Associated Impairment of Sperm Function, Spermatogenesis and Steroidogenesis.

Infection and inflammation are relevant entities of male reproductive disorders that can lead to sub-/infertility. Associated damage of the testis of affected men and in rodent models include leukocytic infiltration, edema formation, fibrosis, germ cell loss and reduced androgen levels. Negative effects on spermatogenesis are thought to be elicited by oxidative stress sustained mostly by increased levels of ROS and pro-inflammatory cytokines. Under normal conditions these cytokines have physiological functions. However, increased levels as seen in inflammation and infection, but also in obesity and cancer are harmful for germ cells and impair steroidogenesis. As a summary, there is mounting evidence that the activation of inflammatory pathways is a rather common feature in various forms of male testicular disorders that extends beyond established infectious/inflammatory cues. This mini review will focus on relevant entities and the mechanisms of how a dysbalance of local testicular factors contributes to disturbances of spermatogenesis and steroidogenesis.

An in-depth analysis of nucleation and growth mechanism of CdS thin film synthesized by chemical bath deposition (CBD) technique.

The aim of this study is to acquire a deeper understanding of the response mechanism that is associated with the formation of CdS thin films. We presented an effective and new hybrid sensitisation technique, which involved the 1-step linker between the related chemical bath deposition (CBD) process and the traditional doping method during CBD for synthesising high-quality, CdS thin films. The mechanism for the combined synthesis of the films is also describes. CdS films were electrostatically bonded to soda-lime glass, causing the formation of the intermediate complexes [Cd(NH3)4]2+, which aided in the collision of these complexes with a soda-lime glass slide. In the one-step fabrication technique, 3-Mercaptopropionic Acid (MPA) was employed as a second source of sulphur ions and a linker molecule. Optical studies showed that the bandgap ranged between (2.26-2.52) eV. CdS + MPA films exhibited a uniform distribution of spherical molecules based on their morphological properties. After annealing, this approach significantly altered the electrical characteristics of CdS films. The CdS + MPA films displayed the highest carrier concentration whereas the CdS + Ag + MPA films exhibited the lowest resistivity, with a jump of 3 orders of magnitude.

New systematic study approach of green synthesis CdS thin film via Salvia dye.

In this study, we aimed to increase the knowledge regarding the response mechanisms which were associated with the formation of CdS thin films. CdS thin film remains the most appealing alternative for many researchers, as it has been a capable buffer material for effect in film based polycrystalline solar cells (CdTe, CIGSe, CZTS). The Linker Assisted and Chemical Bath Deposition (LA-CBD) technique, which combines the Linker Assisted (LA) technique and the chemical bath deposition (CBD) method for forming high quality CdS thin film, was presented as an efficient and novel hybrid sensitization technique. CdS films were bound to soda lime with the help of electrostatic forces, which led to the formation of the intermediate complexes [Cd (NH3)4]2+ that helped in the collision of these complexes with a soda lime slide. Salvia dye and as a linker molecule 3-Mercaptopropionic acid (MPA) was used in the one step fabrication technique. Optical results showed that the bandgap varied in the range of (2.50 to 2.17) eV. Morphological properties showed a homogeneous distribution of the particles that aspherical in shape in the CdS + MPA + Salvia dye films. This technique significantly affected on the electrical characterizations of CdS films after the annealing process. The CdS + Ag + MPA + Salvia dye films showed the maximum carrier concentration and minimum resistivity, as 5.64 × 10 18 cm-3 and 0.83 Ω cm respectively.

Prevalence of Keratoconus and Keratoconus Suspect among Patients Seeking Refractive Surgery in Syria.

PURPOSE: The purpose of this study was to determine the prevalence of keratoconus (KC) and keratoconus suspect (KCS) among patients seeking refractive surgery in Syria. METHODS: This is a retrospective multicenter screening study. The study was conducted in Damascus University, Tishreen University, and Tartous Specialist Eye Center (a private center). Data were collected from refractive surgery candidates referred for preoperative evaluation before laser in situ keratomileusis, photorefractive keratectomy, intrastromal corneal rings, and phakic intraocular lens implantation. Corneal parameters were obtained by Scheimpflug-Placido tomography, Sirius (CSO, Italy). RESULTS: A total of 1479 patients were included in this analysis. The prevalence rates of KC and KCS were 18.19% (269/1479) and 13.52% (200/1479), respectively. In addition, patients with KC were found to have higher percentages of eye rubbing and astigmatism than suspect and normal (P < 0.0001). CONCLUSION: High prevalence rate of KC was found among patients seeking refractive surgery in Syria. Although this study was carried out on a highly selective population, it may reflect a high prevalence rate in a general population in Syria.

Herbal Medicine for Cardiovascular Diseases: Efficacy, Mechanisms, and Safety.

Cardiovascular diseases (CVDs) are a significant health burden with an ever-increasing prevalence. They remain the leading causes of morbidity and mortality worldwide. The use of medicinal herbs continues to be an alternative treatment approach for several diseases including CVDs. Currently, there is an unprecedented drive for the use of herbal preparations in modern medicinal systems. This drive is powered by several aspects, prime among which are their cost-effective therapeutic promise compared to standard modern therapies and the general belief that they are safe. Nonetheless, the claimed safety of herbal preparations yet remains to be properly tested. Consequently, public awareness should be raised regarding medicinal herbs safety, toxicity, potentially life-threatening adverse effects, and possible herb-drug interactions. Over the years, laboratory data have shown that medicinal herbs may have therapeutic value in CVDs as they can interfere with several CVD risk factors. Accordingly, there have been many attempts to move studies on medicinal herbs from the bench to the bedside, in order to effectively employ herbs in CVD treatments. In this review, we introduce CVDs and their risk factors. Then we overview the use of herbs for disease treatment in general and CVDs in particular. Further, data on the ethnopharmacological therapeutic potentials and medicinal properties against CVDs of four widely used plants, namely Ginseng, Ginkgo biloba, Ganoderma lucidum, and Gynostemma pentaphyllum, are gathered and reviewed. In particular, the employment of these four plants in the context of CVDs, such as myocardial infarction, hypertension, peripheral vascular diseases, coronary heart disease, cardiomyopathies, and dyslipidemias has been reviewed, analyzed, and critically discussed. We also endeavor to document the recent studies aimed to dissect the cellular and molecular cardio-protective mechanisms of the four plants, using recently reported in vitro and in vivo studies. Finally, we reviewed and reported the results of the recent clinical trials that have been conducted using these four medicinal herbs with special emphasis on their efficacy, safety, and toxicity.

Western diet aggravates neuronal insult in post-traumatic brain injury: Proposed pathways for interplay.

Traumatic brain injury (TBI) is a global health burden and a major cause of disability and mortality. An early cascade of physical and structural damaging events starts immediately post-TBI. This primary injury event initiates a series of neuropathological molecular and biochemical secondary injury sequelae, that last much longer and involve disruption of cerebral metabolism, mitochondrial dysfunction, oxidative stress, neuroinflammation, and can lead to neuronal damage and death. Coupled to these events, recent studies have shown that lifestyle factors, including diet, constitute additional risk affecting TBI consequences and neuropathophysiological outcomes. There exists molecular cross-talk among the pathways involved in neuronal survival, neuroinflammation, and behavioral outcomes, that are shared among western diet (WD) intake and TBI pathophysiology. As such, poor dietary intake would be expected to exacerbate the secondary damage in TBI. Hence, the aim of this review is to discuss the pathophysiological consequences of WD that can lead to the exacerbation of TBI outcomes. We dissect the role of mitochondrial dysfunction, oxidative stress, neuroinflammation, and neuronal injury in this context. We show that currently available data conclude that intake of a diet saturated in fats, pre- or post-TBI, aggravates TBI, precludes recovery from brain trauma, and reduces the response to treatment.

Y-Shaped Maze to Test Spontaneous Object Recognition and Temporal Order Memory After Traumatic Brain Injury.

Traumatic brain injury (TBI) is one of the most frequent causes of brain damage. Cognitive deficits have been reported in the literature after mild-to-severe TBI affecting memory, language, executive functions, attention, and information processing speed. In this chapter, we describe a method to characterize cognitive impairment in rats following TBI of various intensities. The focus will be on spontaneous object recognition and temporal order memory in rats. These tests are performed in a Y-shaped maze. We have previously identified using this method persistent spontaneous object recognition and temporal order memory deficits following mild-to-moderate TBI in the animals up to 35-day postinjury.

Rodent Models of Methamphetamine Misuse: Mechanisms of Methamphetamine Action and Comparison of Different Rodent Paradigms.

Methamphetamine (METH) is among the most widely used illegal forms of amphetamine. Whether it is injected, snorted, or smoked, METH is a highly addictive substance that affects both peripheral and central nervous system actions. METH use ranges from episodes of binge to chronic use. To investigate METH effects, several animal models have been developed and described to model the various patterns of human METH use. In this work, we examine the molecular, cellular, and structural mechanisms of METH use. Then, we describe the different animal models of METH misuse. Next, we discuss, in details, the acute exposure model which assesses the immediate effects of METH on the brain and the chronic exposure model which best describes the more common long-term consequences of METH use observed in humans. Finally, we tackle the effects and consequences of each paradigm and focus on METH-induced neurotoxic effects and the behavioral changes attributed to each of the described paradigms.

Anti-Inflammatory, Antioxidative, and Hepatoprotective Effects of Trans Δ9-Tetrahydrocannabinol/Sesame Oil on Adjuvant-Induced Arthritis in Rats.

Rheumatoid arthritis (RA) is a painful chronic autoimmune disease affecting the joints. Its first-line therapy, Methotrexate (MTX), although effective in ameliorating the progress of the disease, induces hepatotoxicity over long-term usage. Thus, seeking natural compounds with fewer side effects could be an alternative therapeutic approach. This study aimed to investigate the anti-inflammatory, antiarthritic, and antioxidative effects of synthetic trans-Δ9-tetrahydrocannabinol (Δ9-THC) dissolved in sesame oil (Dronabinol) against MTX in adjuvant-induced arthritis (AIA) rat model. Daily oral administration of Δ9-THC/sesame oil, over a period of 21 days, was well tolerated in arthritic rats with no particular psychoactive side effects. It markedly attenuated the severity of clinical manifestations, recovered the histopathological changes in tibiotarsal joints, and repressed the splenomegaly in arthritic rats. Δ9-THC/sesame oil therapy showed similar effects to MTX in neutralizing the inflammatory process of AIA, through attenuating erythrocyte sedimentation rate (ESR) scores and proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin 1-beta (IL-1ß), and interleukin-6 (IL-6) levels, to normal values. As opposed to MTX, this natural combination markedly protected the liver of arthritic rats and downregulated the induced oxidative stress by increasing the antioxidant defense system such as activities of catalase and superoxide dismutase (SOD) and levels of glutathione (GSH). These results suggest promising effects for the clinical use of Δ9-THC/sesame oil therapy in alleviating arthritic clinical signs as well as arthritis-induced liver injury.

Microwave synthesis, crystal structure, antioxidant, and antimicrobial study of new 6-heptyl-5,6-dihydrobenzo[4,5]imidazo[1,2-c]quinazoline compound.

BACKGROUND: Although the development of antibiotic and antioxidant manufacturing, the problem of bacterial resistance and food and/or cosmetics oxidation still needs more efforts to design new derivatives which can help to minimize these troubles. Benzimidazo[1,2-c]quinazolines are nitrogen-rich heterocyclic compounds that possess many pharmaceutical properties such as antimicrobial, anticonvulsant, immunoenhancer, and anticancer. RESULTS: A comparative study between two methods, (microwave-assisted and conventional heating approaches), was performed to synthesise a new quinazoline derivative from 2-(2-aminophenyl)-1H-benzimidazole and octanal to produce 6-heptyl-5,6-dihydrobenzo[4,5]imidazo[1,2-c]quinazoline (OCT). The compound was characterised using FTIR, 1H and 13C NMR, DIMS, as well as X-ray crystallography. The most significant peak in the 13C NMR spectrum is C-7 at 65.5 ppm which confirms the cyclisation process. Crystal structure analysis revealed that the molecule grows in the monoclinic crystal system P21/n space group and stabilised by an intermolecular hydrogen bond between the N1-H1A…N3 atoms. The crystal packing analysis showed that the molecule adopts zig-zag one dimensional chains. Fluorescence study of OCT revealed that it produces blue light when expose to UV-light and its' quantum yield equal to 26%. Antioxidant activity, which included DPPH· and ABTS·+ assays was also performed and statistical analysis was achieved via a paired T-test using Minitab 16 software with P < 0.05. Also, the antimicrobial assay against two Gram-positive, two Gram-negative, and one fungus was screened for these derivatives. CONCLUSIONS: Using microwave to synthesise OCT have drastically reduced reaction time, and increased yield. OCT show good antioxidant activity in one of the tests and moderate antimicrobial activity.

Measurement of antioxidant activity and antioxidant compounds under versatile extraction conditions: II. The immuno-biochemical antioxidant properties of black sour cherry (Prunus cerasus) extracts.

Retrospectively, we have measured the antioxidant activity and a variety of antioxidant compounds under versatile extraction conditions of sweet cherry (Prunus avium) extracts. Further in this study, in order to understand the biochemical constituents and antioxidant activities of a variety of extracts of black sour cherries (P. cerasus), a related species, antioxidant compounds, including L-ascorbic acid (vitamin C), phenols, flavonoids, and anthocyanins, and the total antioxidant activity were simultaneously measured under varying extraction conditions (mild heating and brief microwave exposure) for: i) whole juice extracts (WJE), ii) methanol-extracted juice (MEJ), iii) ddH2O-extracted pomace (dPOM), and iv) methanol-extracted pomace (mPOM). The antioxidant activity for WJE was substantially increased with mild and prolonged exposure to either heating or microwave, such that the % inhibition against 2,2-diphenyl-1-bspicrylhydrazyl (DPPH) followed a positive correlation (heating, 5-20 min.; microwave, 1-2 min.), insignificant with MEJ and dPOM, whereas with mPOM there was sharp downregulation. L-Ascorbic acid content was not affected with mild to prolonged heating or microwave exposure (WEJ and mPOM), except a mild increase with MEJ and dPOM. Similarly, total phenols assessed showed no significant variations, as compared with control extracts, except a mild decrease with exposure for mPOM. In a manner similar to L-ascorbic acid, total flavonoid content was increased under varying conditions for WEJ and MEJ, and slightly decreased for dPOM and mPOM. On the other hand, anthocyanins showed differential variations with exposure (up- and downregulation). Assessment of extraction means as compared with WJE revealed sharp increase in the antioxidant activity for MEJ, dPOM and mPOM, significant increase in L-ascorbic acid, total phenol, and flavonoid contents for MEJ, dPOM and mPOM, and mild decrease in anthocyanin contents for MEJ, dPOM, and mPOM. These results substantiate the measurable antioxidant activities and contents of P. cerasus extracts under versatile conditions of mild exposure, an effect bearing significant fluctuation with biochemical properties. Since many of those molecules are known to have immuno-biochemical constituencies, antioxidant compounds in sour cherries may have putative antiinflammatory potential and applications in medicinal chemistry, corroborating the observation of regulating and attenuating the growth of microorganisms of medical importance in vitro.

Measurement of antioxidant activity and antioxidant compounds under versatile extraction conditions: I. the immuno-biochemical antioxidant properties of sweet cherry (Prunus avium) extracts.

Previously, we have meticulously examined the efficacy of the measurable antimicrobial activity of sweet cherry (Prunus avium) extracts on a wide spectrum of gram-positive and gram-negative bacteria, in addition to the fungus, Candida albicans, a priori. In order to further understand the biochemical constituents and antioxidant activities of a variety of extracts of sweet cherries, antioxidant compounds of immunological significance, including L-ascorbic acid (vitamin C), phenols, flavonoids, and anthocyanins, and the total antioxidant (free radical scavenging) activity were simultaneously measured under varying and versatile extraction conditions (mild heating [5, 10 and 20 min.], and brief microwave exposure [1, 2 and 5 min.]) for a variety of extracts: i) whole juice extracts (WJE), ii) methanol-extracted juice (MEJ), iii) ddH2O-extracted pomace (dPOM), and iv) methanol-extracted pomace (mPOM). The antioxidant activity under the versatile extraction conditions adopted in this study was conspicuously reduced, such that the % inhibition against 2,2- diphenyl-1-picrylhydrazyl (DPPH) followed an inverse, negative correlational trendline. Moreover, ascorbic acid content was not affected with mild to prolonged heating or microwave exposure, except tangibly with dPOM and mPOM. The total phenols content assessed showed no significant variations, as compared with control extracts. In a manner similar to ascorbic acid, total flavonoids were mildly reduced under varying conditions, an effect mimicked to a certain extent with anthocyanins. Assessment of extraction means as compared with WJE revealed sharp decrease in the antioxidant activity for dPOM and mPOM, significant increase in L-ascorbic acid, total phenol, and flavonoid contents for MEJ, dPOM, and mPOM, and mild decrease in anthocyanin contents for dPOM and mPOM. These results confirm the measurable antioxidant activities and contents of P. avium extracts under versatile conditions of mild exposure, an effect bearing significant biochemical properties of a variety of extraction methods. Further studies are currently investigating the effect of specific antioxidants of P. avium on microbial growth in vitro per se. Since many of the aforementioned molecules hold immunobiochemical constituencies, antioxidant compounds in sweet cherries may have putative anti-inflammatory potential in medicinal chemistry, corroborating the observation of regulating/attenuating the growth of microorganisms of medical importance in vitro.