Untreated Media Research Articles

E-cigarette Vapor Extract Alters Human Eosinophil Gene Expression in an Effect Mediated by Propylene glycol, Glycerin, and Nicotine.

E-cigarette use has become widespread, and its effects on airway inflammation and disease are not fully delineated. E-cigarette vapor extract (EVE) profoundly affects neutrophil function. We hypothesized that EVE also alters eosinophil function and thus could impact allergic airways disease. We employed RNA-sequencing to measure the ex vivo effect of EVE components on human eosinophil transcription. Blood eosinophils from 9 non-vaping subjects without asthma were isolated by negative selection. Cells were incubated for 48 hours with EVE consisting of glycerin, propylene glycol and nicotine (EVE+), EVE without nicotine ("EVE-"), air-exposed media termed Extract Buffer (EB), or untreated media. Bulk RNA-sequencing was performed. Transcriptomic analysis revealed that the EB, EVE-, and EVE+ conditions showed highly variable gene expression with respect to No Treatment and each other. Differential gene expression analysis comparing a combination of EVE+, EVE-, and EB revealed 3,030 differentially expressed genes (DEG) with adjusted p value < 0.05 and log2 fold change >0.5 or <0.5. There were 645 DEG between EB and EVE-, 1,713 between EB and EVE+, and 404 between EVE- and EVE+. Gene set enrichment analysis demonstrated that DEG between both EVE+ and EVE- and the EB control were positively enriched for heme metabolism and apoptosis and negatively enriched TNFα signaling, IFNγ signaling, and inflammatory response. Thus, EVE significantly alters eosinophil metabolic and inflammatory pathways, mediated by propylene glycol and glycerin with both enhancing and unique effects of nicotine. This study motivates further research into the pathogenic effects of vaping on airway eosinophils and allergic airways disease.

Host-Implant Interaction Metabolite 10-HOME Mediated Dysregulation of Adiponectin Resulting in Breast Implant Associated Systemic Manifestations

Background:Breast implant illness (BII) is a poorly understood systemic complication with unknown etiology. Surgical injury at the implant site initiates local inflammation, impacting adipose tissue through decreased adipose-derived adipokine expression. This acute stress responses activates lipid peroxidation pathways, generating oxylipins, causing inflammatory, nociceptive, and vascular responses to injury. One such oxylipin, (E)-10-hydroxy-8-octadecenoic acid (10-HOME), formed from oleic acid, is abundant in host-implant interaction. However, it is unclear how adipose inflammation is maintained. This study aims to elucidate how host-implant interactionmetabolites effect adiponectin expression, subsequently affecting T-cell differentiation eliciting an autoimmune state.&#x0D; Methods:LiSa-2 (human derived adipose tissue secondary cell line) were grown in IMDM and RPMI (4:1) until confluent, then cultured in a well plate. Cultured cells were treated with 10μM 10-HOME for 48h, then were co-cultured with naïve T-cells (PBMC derived) for 48h. T-cells were harvested and flowcytometry was performed using CD4, CD184, CD194, CD196 markers and respective isotypes to verify differentiation (Th1, Th2, Th9/22). To study adiponectin gene expression, treated Lisa-2 cells were harvested for qRT-PCR. ELISA was performed using treated LiSa-2 culture media.&#x0D; Results:We showed decreased expression of adiponectin, after 10-HOME treatment of LiSa-2 cells, compared to untreated cells. qRT-PCR (p=0.0022) showed downregulation of adiponectin gene transcripts in treated LiSa-2 cells, highlighting changes in modulation. Using ELISA (p=0.0006), the culture media of treated cells showed significantly less adiponectin than untreated media. Treated LiSa-2 cells led to polarization of naïve CD4+ T cells to Th1 subtype, evaluated by flowcytometry (p=0.0494). No significant difference in polarization was observed for Th2, Th9 and Th22 subtypes.&#x0D; Conclusion:This study provides evidence that LiSa-2 with 10-HOME treatment caused increased Th1 polarization via modulation of adiponectin expression. Adiponectin regulates many inflammatory pathways, but its effect on Th-cell-mediated responses is poorly understood. Further studies should investigate the mechanism for adiponectin modulation causing T-cell imbalance.

The Effect of SkQ1 on the Oxidative Stress and Inflammatory Responses in Corneal Endothelial Cells During Ex Vivo Expansion

Purpose: The purpose of this study was to characterize the oxidative stress and inflammatory responses in ex vivo corneal endothelial cells (evCEnC) during expansion and assess the impact of SkQ1, an antioxidant and anti-inflammatory compound, on measures of oxidative stress and inflammation. Methods: A CEnC line (HCEnC-21T) was cultured in media supplemented with varying SkQ1 concentrations to determine the optimal SkQ1 dose range of toxicity and protective effect on CEnC exposed to acute oxidative stress. The impact of SkQ1 treatment on intracellular free radical (IFR) levels, NRF2-mediated oxidative stress response, and NFkB-mediated inflammatory response was determined at each passage of evCEnC isolated from donor corneas and cultured in SkQ1-supplemented and untreated media. Results: HCEnC-21T cultured in media supplemented with ≤250 nM SkQ1 retained over 95% cell viability compared with untreated cells. SkQ1 provided oxidative stress protection to HCEnC-21T in a dose-dependent manner up to 500 nM. In evCEnC, 50 nM and 250 nM SkQ1 supplementation significantly reduced IFR levels across passages 0 to 3 (P-values of 0.015 and 0.023, respectively) and 50 nM SkQ1 supplementation led to decreased levels of active NRF2 in evCEnC at passage 2. However, media supplementation with SkQ1 (50 nM and 250 nM) did not alter NFkB activation at any passage. Conclusions: SkQ1 media supplementation provides oxidative stress protection to HCEnC-21T in a dose-dependent manner and decreases IFR levels and NRF2 activation in evCEnC during expansion at concentrations that do not negatively affect CEnC viability. These findings indicate that SkQ1 supplementation may increase the expansion potential of evCEnC.

Anti-Fungal Properties of Plant Extracts in vitro against Alternaria solani, the Cause of Early Blight in Potato

Early blight, caused by Alternaria solani is one of the destructive diseases of potato. An in vitro experiment was conducted using the poisoned food technique to evaluate the efficacy of acetone extracts of six plant species on the mycelial growth of A. solani. Plant extracts of Artemisia vulgaris, Melia azedarach, Parthenium hysterophorus, Datura stramonium, and Justicia adhatoda at concentrations of 0.1%, 0.5% and 5% prepared from Soxhlet extraction were mixed well in the molten potato dextrose agar (PDA) media and poured into petri plates (90 mm Ø). PDA media treated with a fungicide mancozeb was positive control while the untreated media was negative control. The poisoned plates were inoculated with a 4 mm diameter agar plug of seven days old actively growing A. solani and incubated at 26 ± 2 °C, The radial mycelial growth was recorded after 3rd, 5th, 7th and 9th days of incubation and percentage inhibition growth was calculated by comparing it with negative control plates. Artemisia vulgaris significantly inhibited colonial growth at 0.1% and 0.5% concentrations by 36.07% and 12.48%, respectively, at 5th day of inoculation (dai) as compared to negative control. M. azedarach (0.72%), P. hysterophorus (10.17%) and D. stramonium (10.20%) were moderately inhibitory in effect at 1% concentration while J. adhatoda did not possess inhibitory effect on the mycelial growth of A. solani at 5th dai. All the tested extracts were found with some antifungal activity at 5% concentration. A. vulgaris can be exploited as a potent alternative to fungicides for controlling A. solani.

PERICARDIAL INJECTION OF MICRONIZED MATRIX BIOMATERIAL FOR POST-INFARCT CARDIAC REPAIR

BACKGROUND Injection of micronized biomaterials into the pericardial space is a potential less invasive approach to enhance post-infarct cardiac repair (Image). We have shown that porcine small intestinal submucosal extracellular matrix (SIS-ECM) biomaterials contain fibroblast growth factor-2 (FGF-2); and epicardial application of a SIS-ECM patch over ischemic myocardium promotes reparative post-infarct cardiac remodeling by upregulating vasculogenesis and downregulating fibrosis. Micronizing SIS-ECM into an injectable platform will allow pericardial delivery without the need for invasive surgery. Herein, we demonstrate improved post-infarct cardiac recovery after delivering micronized SIS-ECM powder into the pericardial space in in mice and we explore cellular mechanisms using three-dimensional fibroblast cultures. METHODS AND RESULTS In an intact pericardium mouse infarct model, coronary ligation (infarct) was performed through an unopened/undisrupted pericardium followed by pericardial injection of micronized SIS-ECM suspended in saline (treatment; n=12) or saline (control; n=10). Pressure-volume loops assessed cardiac function after 28 days. Mice receiving pericardial injection of micronized SIS-ECM had higher ejection fraction, lower ventricular stiffness (end diastolic pressure volume relationship; EDPVR), higher stroke work and higher cardiac output compared to saline control. Improved diastolic function represented by ventricular stiffness is suggestive of attenuated fibrosis and improved ventricular compliance. Three-dimensional cell cultures of mouse 3T3 cell line fibroblasts in collagen matrices assessed cellular response when exposed to either biomaterial-conditioned media with eluted growth factors (treatment) or untreated media (control). Collagen gels contract when fibroblasts take on a profibrotic phenotype, reducing in size on a macroscopic level. Additionally, paracrine activity of fibroblasts is measured using multiplex analysis with focus on angiogenic VEGF protein and fibrotic MMP-2 protease. Collagen gel contraction and MMP-2 release were attenuated by biomaterial factors, suggesting reduced fibrotic activity. Micronized SIS-ECM increased VEGF production from fibroblasts. Adding an FGF-2 inhibitor to biomaterial-conditioned media negated the biomaterial's effects, thereby increasing gel contraction and MMP-2 release, highlighting the key role of FGF-2 in attenuating fibroblast activity. Table 1 contains outcomes from all studies. CONCLUSION Pericardial injection of micronized SIS-ECM promotes reparative fibroblast activity, increases production of angiogenic VEGF protein, and preserves post-infarct cardiac compliance and function. Pericardial injection is a less invasive modality that may facilitate early intervention to attenuate maladaptive post-infarct structural remodeling. Future study into mechanism and large animal infarct models will be required to identify key cellular pathways and develop percutaneous strategies for clinical translation. Injection of micronized biomaterials into the pericardial space is a potential less invasive approach to enhance post-infarct cardiac repair (Image). We have shown that porcine small intestinal submucosal extracellular matrix (SIS-ECM) biomaterials contain fibroblast growth factor-2 (FGF-2); and epicardial application of a SIS-ECM patch over ischemic myocardium promotes reparative post-infarct cardiac remodeling by upregulating vasculogenesis and downregulating fibrosis. Micronizing SIS-ECM into an injectable platform will allow pericardial delivery without the need for invasive surgery. Herein, we demonstrate improved post-infarct cardiac recovery after delivering micronized SIS-ECM powder into the pericardial space in in mice and we explore cellular mechanisms using three-dimensional fibroblast cultures. In an intact pericardium mouse infarct model, coronary ligation (infarct) was performed through an unopened/undisrupted pericardium followed by pericardial injection of micronized SIS-ECM suspended in saline (treatment; n=12) or saline (control; n=10). Pressure-volume loops assessed cardiac function after 28 days. Mice receiving pericardial injection of micronized SIS-ECM had higher ejection fraction, lower ventricular stiffness (end diastolic pressure volume relationship; EDPVR), higher stroke work and higher cardiac output compared to saline control. Improved diastolic function represented by ventricular stiffness is suggestive of attenuated fibrosis and improved ventricular compliance. Three-dimensional cell cultures of mouse 3T3 cell line fibroblasts in collagen matrices assessed cellular response when exposed to either biomaterial-conditioned media with eluted growth factors (treatment) or untreated media (control). Collagen gels contract when fibroblasts take on a profibrotic phenotype, reducing in size on a macroscopic level. Additionally, paracrine activity of fibroblasts is measured using multiplex analysis with focus on angiogenic VEGF protein and fibrotic MMP-2 protease. Collagen gel contraction and MMP-2 release were attenuated by biomaterial factors, suggesting reduced fibrotic activity. Micronized SIS-ECM increased VEGF production from fibroblasts. Adding an FGF-2 inhibitor to biomaterial-conditioned media negated the biomaterial's effects, thereby increasing gel contraction and MMP-2 release, highlighting the key role of FGF-2 in attenuating fibroblast activity. Table 1 contains outcomes from all studies. Pericardial injection of micronized SIS-ECM promotes reparative fibroblast activity, increases production of angiogenic VEGF protein, and preserves post-infarct cardiac compliance and function. Pericardial injection is a less invasive modality that may facilitate early intervention to attenuate maladaptive post-infarct structural remodeling. Future study into mechanism and large animal infarct models will be required to identify key cellular pathways and develop percutaneous strategies for clinical translation.

Concentration-Dependent Feeding Deterrence to 20-Hydroxyecdysone for Three Subterranean Termite Species (Blattodea: Rhinotermitidae).

Simple SummarySubterranean termite colonies can be eliminated using baiting systems. However, for a given bait to be effective, the active ingredient must be lethal at concentrations that are also palatable to termites. The insect molting hormone, 20-hydroxyecdysone (20E), has potential for use in termite baits, but its palatability to termites has not been examined. The purpose of this study was to determine what concentrations of 20E, if any, cause termite workers to feed less readily. To test this, paper disks were treated with various concentrations of 20E. Groups of 1000 termites of three different species; the Formosan, the Asian and the Eastern subterranean termite; were placed in arenas. The termites had the option of following a path to feed on either a paper disk containing the 20E, or an untreated disk, and the amount of paper consumed was then compared. The results showed that the Asian subterranean termite had the least tolerance for the 20E, the Formosan subterranean termite had a reduced tolerance, and the presence of the 20E had no impact on the Eastern subterranean termite.Effective active ingredients in toxicant bait formulations must be non-deterrent to insect feeding behavior at lethal concentrations. This study evaluated feeding deterrence for Coptotermes formosanus Shiraki, C. gestroi (Wasmann), and Reticulitermes flavipes (Kollar) when provided access to cellulose impregnated with various concentrations of the insect molting hormone, 20-hydroxyecdysone (20E). Termites were exposed to 20E concentrations of 200, 500, 1000 and 2000 ppm and to noviflumuron at 5000 ppm in a 24 h choice-test, and the mass of substrate consumption from treated and untreated media pads was compared for each treatment. 20E feeding deterrence was detected at 500, 1000 and 2000 ppm for C. gestroi, and at 2000 ppm for C. formosanus. No significant differences in consumption of treated and untreated substrate was detected at any concentration for R. flavipes. Potential methods for reducing deterrence are discussed.

Application of innovative technology of biolayer interferometry in basic research in the development of new vaccines

One of the effective methods for real-time protein analysis is based on the phenomenon of light interference and is called "biolayer interferometry". The patented Bio-Layer Interferometry (BLI) technology, which underlies ForteBio Octet platform, provides surface interaction analysis for disposable fiber optic biosensors. The article presents the technology of interferometry of the biolayer of BLI molecules, which can be used to determine the kinetics and affinity, for the specific quantitative determination of molecules bound to the biosensor. The purpose of the review was to summarize scientific data on the use of the interferometry technology of the biolayer of BLI molecules in determining the concentration of proteins and studying the kinetics of interaction using Octet systems when creating new vaccines in many ways. BLI biolayer interferometry technology allows protein concentration measurements to be performed without using of labels or auxiliary reagents, even in untreated media. It is characterized by a sensitivity up to several ng/cm3, provides accurate quantitative changes in a few seconds, and not in many hours like traditional methods (ELISA, HPL Coder). The affinity, concentration, and binding kinetics of the studied proteins can be measured in 4- ml drop samples directly on the laboratory table. The technology of interferometry of the biolayer of BLI molecules can be used by researchers to select the most immunogenic epitopes in the molecular structure of a pathogen, it is also possible to conduct studies on the characterization and recognition, diversity and distribution of pathogens, affinity for antibodies, it is possible to characterize the immune response of the host, to study molecular interactions between the pathogen and the host, and to carry out therapeutic and clinical research. Currently, the technology of interferometry of the biolayer of BLI molecules has already been used in fundamental studies of pathogens that cause HIV, herpes, Ebola, influenza A H7N9, Dengue, malaria, Zika virus, diphtheria, tuberculosis, listeriosis, gastroenteritis, respiratory pathology, including the COVID outbreak. -19.

The HIPPO Transducer YAP and Its Targets CTGF and Cyr61 Drive a Paracrine Signalling in Cold Atmospheric Plasma-Mediated Wound Healing

Reactive species play a pivotal role in orchestrating wound healing responses. They act as secondary messengers and drive redox-signalling pathways that are involved in the homeostatic, inflammatory, proliferative, and remodelling phases of wound healing. The application of Cold Atmospheric Plasma (CAP) to the wound site produces a profusion of short- and long-lived reactive species that have been demonstrated to be effective in promoting wound healing; however, knowledge of the mechanisms underlying CAP-mediated wound healing remains scarce. To address this, an in vitro coculture model was used to study the effects of CAP on wound healing and on paracrine crosstalk between dermal keratinocytes and fibroblasts. Using this coculture model, we observed a stimulatory effect on the migration ability of HaCaT cells that were cocultured with dermal fibroblasts. Additionally, CAP treatment resulted in an upregulation of the HIPPO transcription factor YAP in HaCaTs and fibroblasts. Downstream effectors of the HIPPO signalling pathway (CTGF and Cyr61) were also upregulated in dermal fibroblasts, and the administration of antioxidants could inhibit CAP-mediated wound healing and abrogate the gene expression of the HIPPO downstream effectors. Interestingly, we observed that HaCaT cells exhibited an improved cell migration rate when incubated with CAP-treated fibroblast-conditioned media compared to that observed after incubation with untreated media. An induction of CTGF and Cyr61 secretion was also observed upon CAP treatment in the fibroblast-conditioned media. Finally, exposure to recombinant CTGF and Cyr61 could also significantly improve HaCaT cell migration. In summary, our results validated that CAP activates a regenerative signalling pathway at the onset of wound healing. Additionally, CAP also stimulated a reciprocal communication between dermal fibroblasts and keratinocytes, resulting in improved keratinocyte wound healing in coculture.

Effect of drought stress on morphological, anatomical, and physiological characteristics of Cempo Ireng Cultivar Mutant Rice (Oryza sativa l.) strain 51 irradiated by gamma-ray

Drought stress is a factor that affects plant growth and development, both in terms of morphology, anatomy, and physiology. Mutant Oryza sativa L. strain 51 of Cempo Ireng cultivar as the result of gamma-ray irradiation is superior mutant black rice strain which has a faster planting period of 10-20 days than its control and shorter plant height. This study aims to determine the morphological, anatomical, and physiological responses, especially the proline content inside the leaves of mutant black rice strain 51. The study used a completely randomized design (CRD) with the treatment of drought stress using PEG 6000 in Yoshida liquid media. The seedlings were planted for 21 days in untreated media, then treated for 14 days. Observation of morphological characters was carried out by measuring plant height, root length, leaf area, and plant biomass. Observations of anatomical characters were carried out by observing the cross-section of the root. Observation of physiological character was carried out by measuring leaf proline levels. The results showed that drought stress with PEG 6000 inhibited the growth and development of mutant rice strain 51. Drought stress reduces plant height, root length, leaf area, plant biomass and the area of root aerenchyma. Proline leaf content increased significantly at a PEG concentration of 30%. Mutant rice strain 51 showed a tolerant response to drought stress with the significant increased of proline, the increased of root stele diameter and the constant number of metaxylem.

Interplay between Plasmodium falciparum haemozoin and l-arginine: implication for nitric oxide production

BackgroundPlasmodium falciparum haemozoin, a detoxification product of digested haemoglobin from infected erythrocytes, is released into the bloodstream upon schizont rupture and accumulates in leukocytes. High levels of haemozoin correlate with disease severity. Some studies have shown that concentrations of the substrate of inducible nitric oxide synthase (iNOS), l-arginine, as well as nitric oxide are low in patients infected with P. falciparum malaria. The present study investigates, in vitro, the role of P. falciparum haemozoin on nitric oxide production, iNOS expression in macrophages, and the possible interaction between l-arginine and haemozoin.MethodsPlasmodium falciparum haemozoin was obtained from in vitro cultures through magnetic isolation. Phagocytosis of haemozoin by immortalized bone marrow derived macrophages was detected by confocal reflection combined with fluorescence microscopy. Nitrite concentrations in the supernatants was evaluated by Griess assay as a standard indication of nitric oxide production, while iNOS expression was detected on cell extracts by western blotting. Detection of l-arginine in haemozoin-treated or untreated media was achieved by liquid chromatography–tandem mass spectrometry (LC–MS/MS).ResultsHaemozoin synergizes in vitro with interferon-gamma to produce nitric oxide. However, when mouse macrophages were stimulated with haemozoin, a proportional increase of nitric oxide was observed up to 25 μM of haemozoin, followed by a decrease with doses up to 100 μM, when nitric oxide release was completely abrogated. This was not due to reactive oxygen species production, nor to an effect on iNOS activity. Interestingly, when at 24 h, haemozoin-treated macrophages were washed and incubated in fresh medium for further 24 h, the nitric oxide production was restored in a dose–response manner. Similar results were seen when l-arginine-enriched media was used in the stimulation. Moreover, muramyldipeptide, a strong nitric oxide inducer, was unable to activate macrophages to release nitric oxide in the presence of haemozoin-treated medium. By LC–MS/MS a complete depletion of l-arginine was observed in this haemozoin-treated, conditioned medium.ConclusionsIt is proposed that haemozoin interacts with l-arginine reducing its availability for iNOS, and thus decreasing nitric oxide production. The clinical (or pathological) implications of these results are discussed.

Lipid production enhancement in tropically isolated microalgae by azide and its effect on fatty acid composition

Microalgae, as photosynthetic microorganisms, have been referred to as the third generation of biodiesel feedstock due to their ability to accumulate high amounts of lipids in their intracellular bodies. However, the commercialization of microalgal biodiesel is currently not economical due to high production cost. Recently, a novel strategy was found to overcome this issue and, at the same time, boost the accumulation of lipids by the addition of azide to the microalgal culture. Azide was added to study its effect on the growth and accumulation of fatty acids in three strains of tropical microalgae, identified as Acutodesmus obliquus, Desmodesmus maximus, and Chlorella pyrenoidosa, which were isolated from the Hulu Langat River, Selangor, Malaysia. Lipid production of A. obliquus and D. maximus after azide treatment increased by 40 and 20%, respectively. In contrast, the induction of azide led to insignificant lipid accumulation in C. pyrenoidosa, likely because azide targeted the respiration system rather than lipid production, which consequently caused a slight growth retardation. There was a great difference in the types of fatty acids found in azide-treated cells and non-treated cells. In A. obliquus, more saturated fatty acids (SFAs) were found in azide-treated cells, while more polyunsaturated fatty acids (PUFAs) were found in non-treated cells. In C. pyrenoidosa, the induction of lipid accumulation by azide led to the formation of more SFAs and PUFAs. For D. maximus, the cells synthesized more monounsaturated fatty acids (MUFAs) in both azide-treated and untreated media but azide treatment increased an insignificant amount of PUFAs.

Ethanol production using vegetable peels medium and the effective role of cellulolytic bacterial (Bacillus subtilis) pre-treatment.

Background: The requirement of an alternative clean energy source is increasing with the elevating energy demand of modern age. Bioethanol is considered as an excellent candidate to satiate this demand. Methods: Yeast isolates were used for the production of bioethanol using cellulosic vegetable wastes as substrate. Efficient bioconversion of lignocellulosic biomass into ethanol was achieved by the action of cellulolytic bacteria ( Bacillus subtilis). After proper isolation, identification and characterization of stress tolerances (thermo-, ethanol-, pH-, osmo- & sugar tolerance), optimization of physiochemical parameters for ethanol production by the yeast isolates was assessed. Very inexpensive and easily available raw materials (vegetable peels) were used as fermentation media. Fermentation was optimized with respect to temperature, reducing sugar concentration and pH. Results: It was observed that temperatures of 30°C and pH 6.0 were optimum for fermentation with a maximum yield of ethanol. The results indicated an overall increase in yields upon the pretreatment of Bacillus subtilis; maximum ethanol percentages for isolate SC1 obtained after 48-hour incubation under pretreated substrate was 14.17% in contrast to untreated media which yielded 6.21% after the same period. Isolate with the highest ethanol production capability was identified as members of the ethanol-producing Saccharomyces species after stress tolerance studies and biochemical characterization using Analytical Profile Index (API) ® 20C AUX and nitrate broth test. Introduction of Bacillus subtilis increased the alcohol production rate from the fermentation of cellulosic materials. Conclusions: The study suggested that the kitchen waste can serve as an excellent raw material in ethanol fermentation.

Dvostepena fermentacija za proizvodnju mlečne kiseline na destilerijskoj džibri

The aim of this study was to assess utilization of distillery stillage in a two-stage fermentation using Bacillus licheniformis TFUNS and Lactobacillus paracasei NRRL B-4564. In the first stage the stillage was pretreated with B. licheniformis which is an efficient producer of proteolytic enzymes, in order to increase the content of free α-amino nitrogen in the waste substrate. In the subsequent stage the lactic acid (LA) fermentation by L. paracasei was performed. The results of the fermentation of proteolytically pretreated stillage (i.e. of two-stage fermentation) were compared with the untreated stillage (one-stage fermentation). The results have shown that the amount of free α-amino nitrogen in pretreated media was 107 % higher compared to the initial value. The concentration of the LA obtained in the second stage by L. paracasei was 48 % higher than in untreated stillage. In addition, the growth of L. paracasei in pretreated stillage was also better supported compared to that in untreated media. The process enabled economical and sufficient supply of easily assimilative nitrogen sources needed for lactic acid bacteria, thus avoiding addition of costly sources in the media commonly performed in LA production.

Response of the green alga Oophila sp., a salamander endosymbiont, to a PSII‐inhibitor under laboratory conditions

In a rare example of autotroph-vertebrate endosymbiosis, eggs of the yellow-spotted salamander (Ambystoma maculatum) are colonized by a green alga (Oophila sp.) that significantly enhances salamander development. Previous studies have demonstrated the potential for impacts to the salamander embryo when growth of the algae is impaired by exposure to herbicides. To further investigate this relationship, the authors characterized the response of the symbiotic algae (Oophila sp.) alone to the photosystem II (PSII) inhibitor atrazine under controlled laboratory conditions. After extraction of the alga from A. maculatum eggs and optimization of culturing conditions, 4 toxicity assays (96 h each) were conducted. Recovery of the algal population was also assessed after a further 96 h in untreated media. Average median effective concentration (EC50) values of 123 µg L(-1) (PSII yield), 169 µg L(-1) (optical density), and 299 µg L(-1) (growth rate) were obtained after the 96-h exposure. Full recovery of exposed algal populations after 96 h in untreated media was observed for all endpoints, except for optical density at the greatest concentration tested (300 µg L(-1) ). Our results show that, under laboratory conditions, Oophila sp. is generally less sensitive to atrazine than standard test species. Although conditions of growth in standard toxicity tests are not identical to those in the natural environment, these results provide an understanding of the tolerance of this alga to PSII inhibitors as compared with other species.

The Effects of Dexamethasone and Acyclovir on a Cell Culture Model of Delayed Facial Palsy

Pretreatment with antiherpetic medications and steroids decreases likelihood of development of delayed facial paralysis (DFP) after otologic surgery. Heat-induced reactivation of herpes simplex virus type 1 (HSV1) in geniculate ganglion neurons (GGNs) is thought to cause of DFP after otologic surgery. Antiherpetic medications and dexamethasone are used to treat DFP. Pretreatment with these medications has been proposed to prevent development of DFP. Rat GGN cultures were latently infected with HSV1 expressing a lytic protein-GFP chimera. Cultures were divided into pretreatment groups receiving acyclovir (ACV), acyclovir-plus-dexamethasone (ACV + DEX), dexamethasone alone (DEX), or untreated media (control). After pretreatment, all cultures were heated 43°C for 2 hours. Cultures were monitored daily for reactivation with fluorescent microscopy. Viral titers were determined from culture media. Heating cultures to 43°C for 2 hours leads to HSV1 reactivation and production of infectious virus particles (59 ± 6.8%); heating cultures to 41°C showed a more variable frequency of reactivation (60 ± 40%), compared with baseline rates of 14.4 ± 5%. Cultures pretreated with ACV showed lower reactivation rates (ACV = 3.7%, ACV + DEX = 1.04%) compared with 44% for DEX alone. Viral titers were lowest for cultures treated with ACV or ACV + DEX. GGN cultures harboring latent HSV1 infection reactivate when exposed to increased temperatures that can occur during otologic surgery. Pretreatment with ACV before heat provides prophylaxis against heat-induced HSV reactivation, whereas DEX alone is associated with higher viral reactivation rates. This study provides evidence supporting the use of prophylactic antivirals for otologic surgeries associated with high rates of DFP.

Effects of Ginkgo biloba Extract on Cultured Human Retinal Pigment Epithelial Cells under Chemical Hypoxia

Purpose: To investigate the effects of Ginkgo biloba extract (GBE) on expression of hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) in retinal pigment epithelial (RPE) cells under chemical hypoxia.Materials and Methods: RPE cells (ARPE-19) were cultured in either untreated media (control group), media treated with 200 μM cobalt chloride (hypoxia group) or media treated with both 200 μM cobalt chloride and 100 μg/ml GBE (hypoxia + GBE group) for various amounts of time. HIF-1α and VEGF expression were compared between groups. HIF-1α and VEGF messenger RNA (mRNA) were quantified using real-time polymerase chain reaction (PCR). HIF-1α of extracted nuclei and VEGF of the media were quantified using enzyme-linked immunosorbent assay (ELISA). The expression of HIF-1α was also assessed with Western blot and immunocytochemistry.Results: HIF-1α mRNA, VEGF mRNA, HIF-1α and VEGF levels were higher in the hypoxia group compared with the control group; however, real-time PCR revealed decreased expression of HIF-1α mRNA and VEGF mRNA in the hypoxia + GBE group compared with the hypoxia group. In the ELISA, the HIF-1α and VEGF protein concentrations also decreased with GBE treatment. Western blot and immunostaining showed that the intensity of HIF-1α signals in the hypoxia + GBE group was lower than that of the hypoxia group.Conclusion: GBE reduced HIF-1α and VEGF expression in RPE cells cultured under chemical hypoxia in vitro.

Potential of pyrethroid insecticides and plant extracts on fecundity and egg viability of Tribolium castaneum (Herbst)

Context: Sythetic pyrethroid and two botanical may reduce the fecundity and egg viability of red flour beetle, Tribolium castaneum (Herbst) and can be used in grain protection. Objective: To determine the effect of cypermethrin, fenitrothion, Murraya paniculata L. and Jatropha gossypifolia L. plant extracts on fecundity and egg viability of T castaneum. Materials and Methods: Newly hatched T. castaneum larvae were kept in a petridish containing 30 g either fresh or treated flour separately. After pupation the pupae were sieved and sexed by microscopic examination and kept separately for adult emergence. Twenty five pairs (1?: 1?) adults were paired from treated and one pair from untreated media and placed in a glass vial containing 2 g of food either treated or untreated for oviposition in an incubator at 30 ± 0.5°C. Eggs were collected and counted after every three days over a period of 45 days. The eggs laid by the first 15 pairs from each treatment were collected from the experimental vials and kept in separate petridish for hatching. Eggs were regularly observed with a microscope and hatched larvae were counted. Results: The females of all treated media laid less number of eggs than the untreated females. Both insecticides and botanicals affected oviposition significantly (P&lt;0.001) at all doses levels. All the treatments of both the insecticides and botanicals significantly (P &lt; 0.001) reduced the fertility of eggs laid by T. castaneum females in comparison to those of the control. Conclusion: The plant extracts of kamini and jamalgota can effectively reduced the fecundity and egg viability against stored product insect pests with low mamalian toxicity. DOI: http://dx.doi.org/10.3329/jbs.v19i0.13007 J. bio-sci. 19 95-97, 2011

The role of osteocyte apoptosis in cancer chemotherapy‐induced bone loss

Intensive cancer chemotherapy leads to significant bone loss, the underlying mechanism of which remains unclear. The objective of this study was to elucidate mechanisms for effect of the commonly used anti-metabolite methotrexate (MTX) on osteocytes and on general bone homeostasis. The current study in juvenile rats showed that MTX chemotherapy caused a 4.3-fold increase in the number of apoptotic osteocytes in tibial metaphysis, which was accompanied by a 1.8-fold increase in the number of tartrate-resistant acid phosphatase-positive bone resorbing osteoclasts, and a 35% loss of trabecular bone. This was associated with an increase in transcription of the osteoclastogenic cytokines IL-6 (10-fold) and IL-11 (2-fold). Moreover, the metaphyseal bone of MTX-treated animals exhibited a 37.6% increase in the total number of osteocytes, along with 4.9-fold higher expression of the DMP-1 transcript. In cultured osteocyte-like MLO-Y4 cells, MTX treatment significantly increased caspase-3-mediated apoptosis, which was accompanied by the formation of plasma membrane-born apoptotic bodies and an increase in IL-6 (24-fold) and IL-11 (29-fold) mRNA expression. Conditioned media derived from MTX-treated MLO-Y4 cells was twice as strong as untreated media in its capacity to induce osteoclast formation in primary bone marrow osteoclast precursors. Thus, our in vivo and in vitro data suggested that MTX-induced apoptosis of osteocytes caused higher recruitment of DMP-1 positive osteocytes and increased osteoclast formation, which could contribute towards the loss of bone homeostasis in vivo.

Spinosad Induced Cytogenotoxic Effects on the Mosquito Fish, Poecilia reticulata

Background : Spinosad is an insect control product developed initially for agricultural pests. In recent times, the bio-rational compound has gained popularity in the area of mosquito larval control. The cytogenotoxic potential of the naturally derived compound was investigated on the mosquito fish, Poecilia reticulata to assess its compatibility as a potential larvicide for integrated mosquito larviciding. Methods : Blood samples were collected from the gill epithelial cells of Poecilia reticulata exposed in vivo to three concentrations of Spindor dust (60 μgL - 1 , 123 μgL - 1 , 361 μgL - 1 ). The frequencies of micronucleus and other nuclear abnormal cells as well as, normochromatic cells from treated and untreated media were evaluated after sacrificing the fish at sampling times of 1, 3, 7, 14, 21 and 28 days. Results : The induction of micronucleus, nuclear abnormal and normochromatic cells were highly significant (P<0.01; P<0.001). Polychromatic erythrocytes were more sensitive than binucleated cells and this occurred with increasing concentration of the larvicide (P<0.01). The ratio of polychromatic cell to normochromatic cells increased significantly by concentration and time of exposure when compared with control (P<0.05). Conclusion: The naturally derived Spinosad inhibited mitotic division in Poecilia reticulata therefore it was not compatible with the fish species for integrated mosquito larviciding at the exposed concentrations.

Effects of Vermicompost Tea (Aqueous Extract) on Pak Choi Yield, Quality, and on Soil Biological Properties

This study investigated the effects of vermicompost tea (aqueous extract) on yield and chemical quality of pak choi (Brassica rapa cv Bonsai, Chinensis group) grown in three media (two soils and a peat-perlite medium) under two fertilizer regimes (compost and synthetic fertilizer). The impacts of tea application on the chemical and biological properties of the growth media were also investigated. Vermicompost teas were prepared using various extraction methods (non-aerated, aerated, aerated with additives) with 1:10 (v:v) chicken manure-based vermicompost to water dilution and applied weekly at the rate of 200 mL plant−1 for 4 weeks. Application of vermicompost tea increased plant production, total carotenoids and total glucosinolates in plant tissue. This effect was most prominent under compost fertilization. Total phenolic was lower in vermicompost tea treated plants compared to those treated with only mineral nutrient solution and the water control. Vermicompost tea improved mineral nutrient status of plants and media, and enhanced the biological activity of the media. Variability in yield and chemical quality of plants across treatments was explained largely by variability in tissue N uptake and dry matter accumulation. Dehydrogenase activity and soil respiration of vermicompost tea-treated growth media were approximately 50% higher than untreated media. This study confirmed that vermicompost tea can positively influence plant yield and quality and increase soil biological activity in multiple soil types.