Ethanol-treated Samples Research Articles

Fast-setting multi-stimuli responsive silk hydrogels tailored with dual network interface

Silk fibroin, a naturally occurring and versatile polymer with broad range of material forms and applications, presents numerous advantages. Notably, hydrogels are noteworthy among these advantages due to their inherent hydrophilicity, biocompatibility, and flexibility. Nevertheless, their practical utility has been hindered by intricate preparation procedures and limited mechanical strength. In this investigation, we introduce a straightforward and cost-effective method for producing a novel silk hydrogel. Subsequent to the hydrogel synthesis, an ethanol treatment was employed to induce the preferential formation of β-sheet conformations. The developed system exhibits rapid gelation (gelation time: 3 min) and non-swelling behavior (swelling ratio = 0.47). This suggests its potential suitability for biomedical devices without imposing strain on adjacent tissues. Conformational rearrangement of polymeric chains induced by ethanol treatment contributed to heightened material crystallinity, resulting in a remarkable 55.9% increase in Young's modulus and a substantial 91.2% enhancement in ultimate tensile strength for the ethanol-treated hydrogels (SF/AG/Gly/Eth) when compared to their non-ethanol-treated counterparts (SF/AG/Gly). Additionally, in-situ biodegradation studies on silk hydrogels confirmed the material's degradability, with a notable 94.75% weight loss within 3 days, following pseudo-second-order kinetics (R2 = 0.991). This degradation is attributed to action of proteolytic enzymes on the amino acid units of silk fibroin, alleviating concerns regarding waste generation. Moreover, these pH and water-responsive hydrogels can be harnessed in the fabrication of intelligent biomedical devices. Further, drug release studies on silk hydrogels showed that both ethanol-treated and non-treated samples displayed sustained drug release following the Korsmeyer−Peppas model, with an entrapment efficiency of 65.6% and 48.2%, respectively.

Application of ethanol alleviates heat damage to leaf growth and yield in tomato.

Chemical priming has emerged as a promising area in agricultural research. Our previous studies have demonstrated that pretreatment with a low concentration of ethanol enhances abiotic stress tolerance in Arabidopsis and cassava. Here, we show that ethanol treatment induces heat stress tolerance in tomato (Solanum lycopersicon L.) plants. Seedlings of the tomato cultivar 'Micro-Tom' were pretreated with ethanol solution and then subjected to heat stress. The survival rates of the ethanol-pretreated plants were significantly higher than those of the water-treated control plants. Similarly, the fruit numbers of the ethanol-pretreated plants were greater than those of the water-treated ones. Transcriptome analysis identified sets of genes that were differentially expressed in shoots and roots of seedlings and in mature green fruits of ethanol-pretreated plants compared with those in water-treated plants. Gene ontology analysis using these genes showed that stress-related gene ontology terms were found in the set of ethanol-induced genes. Metabolome analysis revealed that the contents of a wide range of metabolites differed between water- and ethanol-treated samples. They included sugars such as trehalose, sucrose, glucose, and fructose. From our results, we speculate that ethanol-induced heat stress tolerance in tomato is mainly the result of increased expression of stress-related genes encoding late embryogenesis abundant (LEA) proteins, reactive oxygen species (ROS) elimination enzymes, and activated gluconeogenesis. Our results will be useful for establishing ethanol-based chemical priming technology to reduce heat stress damage in crops, especially in Solanaceae.

Ethanol-treated eggshell-derived sorbents for direct air capture

Eggshell is a cheap and environmentally friendly waste stream from the food industry, which could be potentially used for different applications in a circular economy scenario. Carbonation of eggshell derived sorbents has been investigated for calcium looping high temperature applications. Nevertheless, the application of these sorbents for direct air capture is yet to be explored in detail. In this work, waste eggshell (ES) and three different ethanol/water treated eggshell samples (E70, E80 and E90) are assessed for direct air capture and compared to limestone. These samples are exposed to ambient air in two distinct conditions. Namely, i) ambient air at 25 °C in a laboratory and ii) refrigerator conditions at 4 °C in order to simulate how these sorbents might perform in different climatic scenarios for varying geographic areas. Carbonation and hydration conversions were calculated for times of ∼ 3000 h. It was found that treating the eggshells with ethanol was key in order to obtain a suitable material for this application. Two of the ethanol-treated samples obtained similar conversions in comparable amounts of time for ambient air conditions, while limestone still performed better in refrigerator conditions.

Novel attempt at discrimination of a bullet-shaped siphonophore (Family Diphyidae) using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-ToF MS)

One major difficulty in identifying the gelatinous bodied bullet-shaped Siphonophore, Diphyids, is that their shape is deformed following ethanol fixation. Ethanol often is preferred over other fixatives, since samples fixed in ethanol can be used for molecular studies that can supplement morphological findings. To overcome this problem, we obtained protein mass spectra of ten species of Diphyidae found in the waters of the Kuroshio Current (Northwest Pacific and South Coast of South Korea) to test whether MALDI-ToF MS could be used as a methodology for species identification. In addition, a number of morphological characteristics that can be used with ethanol-treated samples was summarized. Concatenated phylogenetic analysis was also performed to determine the phylogenetic relationship by obtaining partial sequences of four genes (mtCOI, 16S rRNA, 18S rRNA, and ITS regions). Based on our integrative analysis, MALDI-ToF MS was evaluated as a potentially fast, inexpensive, and accurate tool for species identification along with conventional morphological and DNA barcoding for Diphyidae.

Optimization Strategies for High Photoluminescence Quantum Yield of Monolayer Chemical Vapor Deposition Transition Metal Dichalcogenides.

Chemical vapor deposition (CVD) is a promising method to obtain monolayer transition metal dichalcogenides (TMDCs) with high quality and enough size to meet the requirements of practical photoelectric devices. However, the as-grown monolayers often exhibit a lower PL performance due to the stress between the as-grown TMDCs flakes and the substrate. Therefore, finding a facile method to effectively promote the photoluminescence quantum yield (PL QY) of CVD monolayer TMDCs with a clean surface is highly desirable for practical applications. In this work, based on the CVD monolayers MoS2 and MoSe2, the effect of various stress relaxation methods on the TMDCs PL enhancement is systemically studied. By comparing the different kinds of volatile solution treatment processes, as well as the traditional transfer process, it can be found that the volatile solution with a moderate volatilization rate such as ethanol or IPA is a preferred option to improve the PL performance of the CVD monolayer TMDCs, which also surpasses the traditional transfer method by avoiding wrinkles, defects, and contamination to the samples. PL QY of ethanol-treated CVD samples could increase by 6 times on average. Significantly, PL QY of CVD MoSe2 treated by ethanol can reach ∼16%, which is at the forefront of the previous reports of 2D MoSe2. Our study demonstrated an optimized method to enhance the PL QY of CVD monolayer TMDCs, which would facilitate TMDCs optoelectronics.

Impact of the surface modifications and cell culture techniques on the biomechanical properties of PDMS in relation to cell growth behavior

PDMS is the most widely used material in the microfluidic systems because of its advantageous properties including ease of manufacture, low cost, transparency, and gas permeability. Although all biomaterials need to be sterilized for usage for in vitro or in vivo systems to avoid possible contaminants, none of them proved effective in providing the gold standards so far for PDMS sterilization. Therefore we evaluated the influence of both UV and ethanol sterilization of PDMS substrates on the cell behavior via investigating cell proliferation, oxidative stress and nutrient metabolism of 293 T cells growing on the PDMS first time in the literature. Modulus of elasticity and stress relaxation of UV-treated PDMS samples increased compared to the ethanol-treated ones leading to a state of surface stiffness preferable for cell growth and proliferation. Oxidative stress metabolism of cells growing on the UV-treated PDMS was more stable compared to the ethanol-treated samples. Trace element and mineral metabolism are not impaired in the cells growing on the UV-treated PDMS samples. First time in the literature, we showed that UV treatment is more suitable for the PDMS samples used in the cell culture experiments.

The Physiochemical Properties and Adsorption Characteristics of Processed Pomelo Peel as a Carrier for Epigallocatechin-3-Gallate.

The NaOH-HCl- and ethanol-pretreated pomelo peel samples were prepared to apply to the batch adsorption for epigallocatechin-3-gallate (EGCG). The characteristics of peel samples were determined by Fourier transform infrared spectroscopy, scanning electron microscopy and a laser particle analyzer. The results of the physiochemical properties of the peel samples demonstrate that these peel samples have a promising adsorption capacity for EGCG, because of the increased potential binding sites on the surface compared with those of untreated peel samples. These two peel samples showed enhanced adsorption capacities of EGCG compared with that of unmodified peel in terms of the isothermal adsorption process, which could be described by both Langmuir and Freundlich models, with the theoretical maximum adsorption capacity of 77.52 and 94.34 mg g−1 for the NaOH-HCl and ethanol-treated peel samples, respectively. The adsorption kinetics demonstrated an excellent fitness to pseudo-second-order, showing that chemisorption was the rate-limiting step. The thermodynamics analysis revealed that the adsorption reaction was a spontaneous and endothermic process. This work highlights that the processed pomelo peels have outstanding adsorption capacities for EGCG, which could be promising candidates for EGCG delivering in functional food application.

Culture dependent and independent analyses suggest a low level of sharing of endospore-forming species between mothers and their children

Spore forming bacteria comprise a large part of the human gut microbiota. However, study of the endospores in gut microbiota is limited due to difficulties of culturing and numerous unknown germination factors. In this study we propose a new method for culture-independent characterization of endospores in stool samples. We have enriched DNA of spore-forming bacterial species from stool samples of 40 mother-child pairs from a previously described mother-child cohort. The samples were exposed to a two-step purification process comprising ethanol and ethidium monoazide (EMA) treatment to first kill vegetative cells and to subsequently eliminate their DNA from the samples. The composition of the ethanol-EMA resistant DNA was characterized by 16S rRNA marker gene sequencing. Operational taxonomic units (OTUs) belonging to the Clostridia class (OTU1: Romboutsia, OTU5: Peptostreptococcaceae and OTU14: Clostridium senso stricto) and one belonging to the Bacillus class (OTU20: Turicibacter) were significantly more abundant in the samples from mothers and children after ethanol-EMA treatment than in those treated with ethanol only. No correlation was observed between ethanol-EMA resistant OTUs detected in children and in their mothers, which indicates that a low level of spore-forming species are shared between mothers and their children. Anaerobic ethanol-resistant bacteria were isolated from all mothers and all children over 1 year of age. Generally, in 70% of the ethanol-treated samples used for anaerobic culturing, 16S rRNA gene sequences of bacterial isolates corresponded to OTUs detected in these samples after EMA treatment. We report a new DNA-based method for the characterization of endospores in gut microbiota. Our method has high degree of correspondence to the culture-based method, although it requires further optimization. Our results also indicate a high turnover of endospores in the gut during the first two years of life, perhaps with a high environmental impact.

Improving the emulsification of soy β-conglycinin by alcohol-induced aggregation

Soy proteins have attracted fast increasing interests to be used in food emulsion formulations, due to their good emulsifying properties. Soy β-conglycinin (SC) and glycinin are the two major storage globulins in soy proteins, with the former generally recognized to exhibit much better emulsifying properties than the latter. The current work reported that the treatment of SC with ethanol at concentrations ([E]) of 20–80% (v/v) remarkably improved its emulsification performance and interfacial stabilization, in a [E]-dependent way. The influence of the ethanol treatment on the physicochemical and structural properties of SC was characterized. As expected, the treatment led to an [E]-dependent aggregation of SC, and formation of larger particles with more compact structure. Next, the emulsifying and interfacial properties of these ethanol-treated SC samples were evaluated in terms of emulsification performance and interfacial stabilization, at two oil volume fractions of 0.2 and 0.8. In both cases, the ethanol-treated SC showed greatly improved emulsification performance and interfacial stabilization. The improvements were largely due to the facilitated formation of bridged emulsions, via the strengthened Pickering stabilization by ethanol-induced aggregation. Even at a low concentration of 0.5 wt%, the ethanol-treated SC could still produce a kind of fine emulsions (produced by microfluidization), or high internal phase emulsions with a self-supporting gel network (that could resist the disruption of 6 M urea). The results demonstrated that the ethanol pretreatment can be applied as an effective modification to remarkably improve the emulsifying properties of SC, through the strengthening of its Pickering stabilization.

A three-dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity.

In this study, we propose the design and fabrication of a liver system on a chip. We first chose the most suitable three-dimensional liver-like model between cell spheroids and microtissue precursors, both based on the use of hepatocellular carcinoma cells (HepG2) to provide proof-of-concept data. Spheroids displayed high cell density but low expression of the typical hepatic biomarkers, whereas microtissue precursors showed stable viability and function over the entire culture time. The two liver-like models were compared in terms of cell viability, function, metabolism, and the P-glycoprotein 1 (P-gp) transport-protein expression with the microtissue precursors showing the best performance. Thus, we cultured them into a microfluidic biochip featured with three parallel channels shaped to mimic the hepatic sinusoids. To assess the detoxification potential of the microtissue-loaded biochip we challenged it with a model molecule (ethanol) at different concentrations and time points. Ethanol cytotoxicity was detected by a noninvasive measurement of cell viability based on cell autofluorescence. As expected, a dose-dependent decrease of albumin and urea secretion was observed in the ethanol-treated samples. We believe that the described totally human-derived platform, suitable for integration into a multiorgan microfluidic system, can provide a consistent innovative platform for drug development and toxicity studies.

Quantitative iTRAQ LC–MS/MS Proteomics Reveals Metabolic Responses to Biofuel Ethanol in Cyanobacterial Synechocystis sp. PCC 6803

Recent progress in metabolic engineering has led to autotrophic production of ethanol in various cyanobacterial hosts. However, cyanobacteria are known to be sensitive to ethanol, which restricts further efforts to increase ethanol production levels in these renewable host systems. To understand the mechanisms of ethanol tolerance so that engineering more robust cyanobacterial hosts can be possible, in this study, the responses of model cyanobacterial Synechocystis sp. PCC 6803 to ethanol were determined using a quantitative proteomics approach with iTRAQ LC-MS/MS technologies. The resulting high-quality proteomic data set consisted of 24,887 unique peptides corresponding to 1509 identified proteins, a coverage of approximately 42% of the predicted proteins in the Synechocystis genome. Using a cutoff of 1.5-fold change and a p-value less than 0.05, 135 and 293 unique proteins with differential abundance levels were identified between control and ethanol-treated samples at 24 and 48 h, respectively. Functional analysis showed that the Synechocystis cells employed a combination of induced common stress response, modifications of cell membrane and envelope, and induction of multiple transporters and cell mobility-related proteins as protection mechanisms against ethanol toxicity. Interestingly, our proteomic analysis revealed that proteins related to multiple aspects of photosynthesis were up-regulated in the ethanol-treated Synechocystis cells, consistent with increased chlorophyll a concentration in the cells upon ethanol exposure. The study provided the first comprehensive view of the complicated molecular mechanisms against ethanol stress and also provided a list of potential gene targets for further engineering ethanol tolerance in Synechocystis PCC 6803.

Ethanol injection induced cavitation and heating in tissue exposed to high intensity focused ultrasound

High Intensity Focused Ultrasound (HIFU) can ablate tumors located deep in the body through highly localized energy deposition and tissue heating at the target location. The volume of a HIFU-induced thermal lesion can be increased in the presence of cavitation. This study explores the effect of ethanol injection on cavitation and heating in tissue-mimicking phantoms and bovine liver tissues exposed to HIFU. The HIFU transducer (0.825 MHz) operated at seven acoustic power levels ranging from 1.3 W to 26.8 W. The cavitation events were quantified by B-mode ultrasound imaging, needle hydrophone measurements, and passive cavitation detection (PCD). Temperature in or near the focal zone was measured by thermocouples embedded in the samples. The onset of inertial cavitation in ethanol-treated phantoms and bovine liver tissues occurred at a lower power level than in the untreated samples (control). The cavitation occurrence in turn resulted in a sudden rise of temperature in ethanol-treated samples at a lower acoustic power than that in control. The results of this work indicate that the use of percutaneous ethanol injection prior to HIFU exposure may improve the HIFU therapeutic efficiency.

Effect of ethanol injection on cavitation and heating of tissues exposed to high-intensity focused ultrasound

Cavitation activity and temperature rise have been investigated in a tissue-mimicking material and excised bovine liver treated with ethanol and insonated with a 0.825 MHz focused acoustic transducer. The acoustic power was varied from 1.3 to 26.8 W to find the threshold leading to the onset of inertial cavitation. Cavitation events were quantified by three independent techniques: B-mode ultrasound imaging, needle hydrophone measurements and passive cavitation detection. Temperature in or near the focal zone was measured by thermocouples embedded in the samples. The results of this study indicate that the treatment of tissue phantoms and bovine liver samples with ethanol reduces their threshold power for inertial cavitation. This in turn leads to a sudden rise in temperature in ethanol-treated samples at a lower acoustic power than that in untreated ones. The analysis of passive cavitation detection data shows that once the threshold acoustic power is reached, inertial cavitation becomes a major contributor to acoustic scattering in ethanol-treated phantoms and bovine liver samples as compared to control. This study opens up the possibility of improved tumor ablation therapy via a combination of percutaneous ethanol injection and high-intensity focused ultrasound.

Cellulose acetate electrophoresis of canine plasma after fibrinogen precipitation by ethanol

In routine canine medicine, anticoagulated blood is often the only sample sent to laboratories for diagnostic purposes. This hampers the interpretation of protein electrophoretic tracings because plasma contains fibrinogen, which migrates in the beta-gamma region. In human medicine, fibrinogen can be precipitated from plasma using ethanol. The purpose of this study was to assess ethanol precipitation as a method for removing fibrinogen from canine plasma so as to facilitate the interpretation of electorphoresis results. Blood samples collected from 40 dogs were divided into plain tubes and tubes containing EDTA (n=20) or lithium-heparin (n=20). An aliquot of plasma from each sample was incubated with ethanol at a final concentration of 100 mL/L. Cellulose acetate electrophoresis was then performed on serum, plasma, and plasma treated with ethanol. To verify the efficiency of ethanol treatment, fibrinogen was added to 5 canine serum samples at final concentrations of 2.5, 5.0, and 10.0 g/L, and electrophoresis was performed before and after ethanol treatment. Visual analysis of electrophoretograms from ethanol-treated samples confirmed the disappearance of the fibrinogen peak from the beta(2)-globulin region. Treatment with ethanol caused a significant decrease in the percentage of beta(2)-globulins and a significant increase in the percentage of alpha(2)-globulins. Absolute values of most electrophoretic fractions were significantly decreased in ethanol-treated plasma compared with serum. Ethanol treatment successfully removed fibrinogen from canine plasma and normalized electrophoretic profiles, but probably also precipitated proteins other than fibrinogen. Ethanol treatment is recommended to facilitate visual identification of abnormal monoclonal peaks, but not for determining absolute protein concentrations in electrophoretic fractions.

Effects of Gangliosides on Ethanol‐Induced Neurodegeneration in the Developing Mouse Brain

Ethanol exposure induces apoptotic neurodegeneration in the developing rodent brain during synaptogenesis. This process has been studied as a model for fetal alcohol syndrome. Previously, we have shown that gangliosides and LIGA20 (a semisynthetic derivative of GM1 ganglioside) attenuate ethanol-induced apoptosis in cultured neurons. In the present study, the effects of GM1 and LIGA20 on ethanol-induced apoptotic neurodegeneration were examined using an in vivo neonatal mouse model. Seven-day-old C57BL/6By (B6By) mice were pretreated twice with intraperitoneal administration of GM1 (30 mg/kg), LIGA20 (2.5 mg/kg), or saline, followed by subcutaneous injection of either saline or ethanol (2.5 g/kg) twice with a 2 hours interval. Then the brains were: (1) perfusion-fixed 24 hours after the first ethanol injection, and the extent of neurodegeneration was assessed by cupric silver staining of the brain sections, or (2) perfusion-fixed 8 hours after the first ethanol injection, and the sections were immunostained with anti-cleaved (activated) caspase-3 antibody to evaluate caspase-3 activation. The comparison of cupric silver stained coronal sections indicates that ethanol-induced widespread neurodegeneration in the forebrains of B6By mice was reduced overall by GM1 and LIGA20 pretreatments. The extent of neurodegeneration detected by silver impregnation and activated caspase-3 immunostaining was quantified in the cingulate and retrosplenial cortices, which were the regions most severely affected by ethanol. The results indicate that GM1 and LIGA20 pretreatments induced statistically significant reductions-approximately 50% of the ethanol-treated samples-in silver impregnation and activated caspase-3 immunostaining. No significant differences were observed between saline controls and samples treated with GM1 or LIGA20 alone. These results indicate that GM1 and LIGA20, which have been shown to be neuroprotective against insults caused by various agents, partially attenuate ethanol-induced apoptotic neurodegeneration in the developing mouse brain.

Use of ethanol for preserving steroid and indoleamine hormones in bird plasma

Endocrinological research on wild animals inhabiting remote areas has been hampered by the need to store plasma samples at subzero temperatures. In an attempt to remedy this logistical issue, we here investigate the use of ethanol as an alternative to freezing for the preservation of steroid and indoleamine hormones in avian plasma. Known quantities of the steroids 5α-dihydrotestosterone (DHT), testosterone, 17β-estradiol, corticosterone, and the indoleamine melatonin were added to a stripped pool of chicken plasma. Samples were either immediately frozen at −40 °C, or treated with pure ethanol. Ethanol-treated samples were either immediately frozen, or—to simulate storage conditions at various field locations—left sitting at room temperature for one to two months, or incubated at 36 °C for one month before all treatment groups were frozen at −40 °C. All samples were then analyzed by radioimmunoassay. For DHT and estradiol there were no differences among treatment groups suggesting that ethanol-treatment is as effective as immediate freezing in preserving plasma steroid concentrations. For testosterone, corticosterone and melatonin ethanol-treated samples differed significantly from immediately frozen samples suggesting that caution is needed when comparing absolute concentrations of hormones between samples preserved in different ways. However, differences among ethanol-treated samples in general were small, demonstrating the feasibility of this preservation method in the field at remote locations.

Identification of a 251‐bp Fragment of the PAI‐1 Gene Promoter That Mediates the Ethanol‐Induced Suppression of PAI‐1 Expression

Moderate alcohol consumption reduces the risk for coronary heart disease. This cardioprotection may be due to ethanol enhancement of fibrinolysis. Fibrinolysis involves the interaction of plasminogen activators (PAs) and the plasminogen activator inhibitor type-1 (PAI-1). Factor(s) that decrease endothelial cell (EC) PAI-1 expression increase fibrinolysis and may decrease the risk for cardiovascular disease. Five promoter deletion fragments were generated from a 1.1-kb PAI-1 promoter fragment and ligated to a luciferase reporter gene. Cultured human umbilical vein endothelial cells (HUVECs) were transiently transfected with these PAI-1 deletion constructs. A 251-base pair (bp) fragment of the PAI-1 promoter, positions -800 to -549, was cloned upstream of a heterologous promoter/enhancer. ECs luciferase activity was measured in the absence/presence of 20 mM ethanol. Electrophoresis mobility shift assays were performed with nuclear extracts from untreated and ethanol-treated ECs using this 251-bp fragment. Deletion analysis showed a region between position -800 and -549 mediated ethanol repression of luciferase activity. This 251-bp promoter fragment also repressed the activity of a heterologous promoter/enhancer in the presence of ethanol. Using the labeled 251-bp fragment, nuclear extracts from ethanol-treated ECs contained two inducible bands and one enhanced band. Non-ethanol treated nuclear extracts also contained a band that was not observed in ethanol-treated samples. Competition using 100-fold molar excess of unlabeled probe abolished these four bands. Repression of PAI-I gene transcription in cultured HUVECs exposed to ethanol may involve the interaction of several transcription factors with binding sites localized between positions -800 and -549 of the PAI-1 gene promoter.

Selective Changes in Protein Kinase C Isoforms and Phosphorylation of Endogenous Substrate Proteins in Rat Cerebral Cortex during Pre- and Postnatal Ethanol Exposure

The effect of pre- and postnatal ethanol exposure on protein kinase C (PKC) activity, immunochemical analysis of PKC α, βI, βII, γ, δ, ϵ, η, and ζ by isoform-specific antibodies, andin vitrophosphorylation of endogenous substrate proteins was investigated in rat cerebral cortex. The PKC activity was increased throughout the development. However, the activity at the age of 8 days was significantly high in cytosolic and membrane fractions from ethanol-treated rats. Immunochemical analysis showed increased levels of PKC βI and βII at the age of 8 days, and a decrease in δ isoform at 8, 30, and 90 days of age. PKC isoforms α, γ, ϵ, and η showed no appreciable change in ethanol-treated rats. PKC ζ levels were high in the cytosolic fraction from ethanol-treated samples of 90 days age.In vitrophosphorylation of endogenous substrate proteins in the presence of Ca2+/phospholipid showed increased phosphorylation of selective membrane and cytosolic proteins with 87, 65, 50, 43, 36, and 29 kDa in ethanol-treated rats. The phosphorylation of these proteins decreased in the presence of staurosporine, which also supported PKC-mediated phosphorylation. Increased PKC activity, activation of βI and βII isoforms, decreased levels of δ isoform, and phosphorylation of selective substrate proteins in cerebral cortex due to alcohol exposure might be relevant in ethanol-induced central nervous system dysfunction and fetal alcohol syndrome.

Microscopic Examination of Chilling Injury Symptoms in Ethanol-treated Cucumber Seedling Roots

Cucumber seedlings were germinated for 24 h at 25 °C and half were immersed in a 500 mM ethanol solution for 2 h. After rinsing, seedlings were chilled for 96 h at 2 °C. Control and ethanol-treated samples were taken for light and electron microscopy immediately before and after chilling, and after re-warming for 24 and 72 h. Preliminary experiments revealed visual chilling symptoms such as pinching of the root in a region just above the root tip. This region was excised under a microscope, fixed, and mounted for microscopic observations. The cortical cells of ethanol-treated seedlings before chilling appeared to be irregular in shape with irregular edges, and some epidermal damage was evident. Chilling caused much more epidermal damage in the control seedlings when compared to the ethanol-treated seedlings. After chilling, cortical cells in the control seedlings were observed to be irregularly shaped while those treated with ethanol had round cells. Upon re-warming, control seedlings exhibited increasing epidermal damage with broken cell walls, while ethanol-treated seedlings exhibited more differentiation in the stele.

Ethanol fixation of sputum sediments for DNA-based detection of Mycobacterium tuberculosis

The effect of ethanol fixation on PCR detection and viability of Mycobacterium tuberculosis in human sputum sediments was evaluated. M. tuberculosis seeded into sputum sediments was efficiently killed when treated for 1 h with 50, 70, or 95% ethanol. PCR amplification of a 123-bp fragment of the M. tuberculosis-specific IS6110 was not affected in ethanol-treated samples even when fixation was extended to 24 h. Ethanol fixation of sputum sediments did not affect the PCR detection of M. tuberculosis in clinical samples. PCR results from ethanol-treated clinical samples containing M. tuberculosis (smear positive and smear negative) or other respiratory pathogens correlated directly with the results by conventional detection methods for M. tuberculosis. Our results show that ethanol fixation of human sputum sediments containing M. tuberculosis significantly reduces the potential exposure of workers to viable M. tuberculosis without affecting DNA analysis by PCR. Also, ethanol fixation of sputum sediments provides a simple and inexpensive way to store and transport clinical specimens identified for DNA-based diagnostics without refrigeration.