Saline Extracts Research Articles

Extraction Of House Dust Mite Allergen, Der P1, From Dust

A recent article in this journal found that the type of extraction buffer did not affect measured levels of the major group one allergen (Der p 1) from the house dust mite, Dermatophagoides pteronyssinus, in household dust samples (1). This differed from a previous study where about twice the levels of Der p 1 were found in borate-buffered saline extracts, compared to ammonium bicarbonate buffer or phosphate-buffered saline extracts, irrespective of extraction temperature or time of extraction (2). In discussing our previous study (2), the authors state that we did not use Tween-20 in our extraction buffers. Although our article did not make a mention of Tween-20, we in fact did use it in our extraction media. It was an oversight of us in that we did not mention this in our article. We further explored temperature and buffer effects on Der p 1 extraction in an external quality control program where six dust samples were sent to 23 laboratories worldwide (3). Results from this study again showed that type of extraction buffer and extraction temperature influenced Der p 1 measurements. The majority of the participant laboratories did use Tween-20 as a wetting agent in their extraction buffers. As pointed out by the authors, adding Tween-20 to extraction media results in higher endotoxin levels and is recommended by the European Committee for Standardisation. The authors call for further investigation on the effects of Tween-20 on extraction effi ciency of all indoor allergens from dust. However, the majority of indoor allergen analysis laboratories use ELISA or MARIATM kits from Indoor Biotechnologies where the recommended extraction medium is phosphate-buffered saline containing 0.02 % Tween-20. It would take much effort to study this systematically to obtain results with suffi cient internal and external validity.

Analysis of mitochondrial DNA damage using 8-hydroxy 2′deoxyguanosine on in vitro and in vivo exposure of biomaterials

The present study was carried out to evaluate the effect of HAP-EVA, fibrin glue, HA-BG, Latex and Dental material on oxidative stress related mtDNA damage by in vitro and in vivo methods. In vivo studies of these biomaterials were carried out by implanting biomaterials (five materials) on animals for period of 1, 4, 12, 26 and 52 weeks. At the end of observations, animals were anesthetized, sacrificed and tissues surrounding the implanted materials were collected. Brain, bone and muscles were used for the extraction of mtDNA. Similarly mtDNA was extracted from the homogenate of fresh brain, bone and muscles on exposure to the physiological saline extract of all the above five biomaterials (In vitro). The extracted mtDNA were subjected to analyse the presence of 8-OHdG. The results of study indicated that there was no significant increase in the level of 8-OHdG and thereby does not influence on the GC-TA transversions.

Purification and Characterization of a Lectin of the Swartzieae Legume Taxa

This work aimed at describing the first biochemical and structural data of a lectin belonging to Swartzieae, a primitive Legume Taxa. A lactose-binding seed lectin (SLL) was purified by affinity chromatography of crude saline extracts of Swartzia laevicarpa on immobilized lactose. The SLL agglutinated rabbit erythrocytes but not rat or human (A, B, O) erythrocytes. Lectin activity was retained after heating at 100 �C for 15 min and was best inhibited by Nacetylgalactosamine, lactose and galactose. The lectin exhibited a single electrophoretic pattern that corresponded to a molecular mass of 29,000 Da, which was confirmed by MS analysis. In addition, the lectin reacted positively with Schiff's reagent. The unique N-terminal amino acid sequence (39 residues) and the internal peptide sequence were determined by Edman degradation and MS/MS, respectively. The sequencing revealed complete homology of the SLL with legume lectins belonging to primitive groups (Dalbergieae and Sophoreae). The SLL (at 1 mg/ml) did not exhibit antifungal activity against various phytopathogens or cytotoxicity (at 100 μg/ml) towards different cancer cell lines.

칡 추출물의 면역세포 활성화 효과

In this experiment, the effects of Pueraria thunbergiana extracts on the activation of immune cells were studied. An immune cell-activating factor was partially purified from P. thunbergiana by means of physiological saline extraction, acetone precipitation, and heating inactivation. P. thunbergiana extracts increased the proliferation of spleen cells and induced the production of IL-2, IL-6, TNF-α, and IFN-γ by spleen cells. Also, they increased the proliferation of purified B cells and the production of IgM antibody in a dose-dependent fashion. The extract self-induced NO synthesis in a mouse macrophage cell line (RAW264.7). When cell lines were treated with extracts, the cytokines’ (IL-1β, IL-6, and TNF-α) production was markedly increased. Therefore, P. thunbergiana extract can self-activate spleen cells, B cells, and macrophages. These results might be useful in further studies into a possible immune-activating agent derived from P. thunbergiana for the development of functional foods and drugs.

Evaluation of tissue reaction to Aroeira (Myracrodruon urundeuva) extracts: a histologic and edemogenic study

ObjectivesThis study evaluated subcutaneous tissue response to Aroeira (Myracrodruon urundeuva) extract employing edemogenic and histological analyses. Material and methodsTest groups consisted of aqueous and ethanolic Aroeira extracts and saline (control). For edema quantification, 18 rats received an intravenous injection of Evan's Blue. After 30 min, the extracts and saline were injected on the dorsum of the rats, which were then sacrificed after 3 and 6 h. Readings were performed in a spectrophotometer. For subcutaneous implantation, 30 rats received a polyethylene tube containing the extracts on their dorsum and then they were killed after 7 and 28 days. The samples were processed for histological analysis and evaluated with a light microscope. The inflammatory infiltrate was quantified. ResultsThere were no statistically significant differences between aqueous extract and saline groups in relation to edema quantification in the different periods (p>0.05). Ethanolic solution resulted in more edema independently of the experimental period (p<0.05). Histological analysis showed similar results on the 7-day period for the 3 groups. There was a notable reduction on inflammatory cell number for saline and aqueous extract groups at 28 days. ConclusionThe aqueous extract showed biocompatible properties similar to those of saline.

Assessment of micronuclei and chromosomal anomalies of five biocompatible materials in mice

In vivo genetic toxicology tests measure direct DNA damage or the formation of gene or chromosomal mutations, and are used to predict mutagenic and carcinogenic potential of compounds for regulatory purposes. These adverse genotoxic effects may be manifested in the form of gene mutations, structural chromosomal aberrations (CA), recombination, and numerical changes. The present investigation was carried to assess genotoxic effects of five different implantable biomaterials developed in different laborataries of Sree Chitra Tirunal Institute of Medical Sciences and Technology. All biomaterials were developed for clinical applications. CA and micronuclei (MN) studies are biomarkers of genotoxicity testing. Leachants from the extract of biomaterials are capable of inducing structural and numerical chromosomal changes. The studies were conducted in Swiss albino mice with the physiological saline extract of materials together with cyclophosphamide and physiological saline as positive and negative controls. Animals were administered intraperitoneally (ip) with a single injection of test, positive (cyclophosphamide), and negative (physiological saline) control and sacrificed after 24 or 48 h. Bone marrow cells were collected for CA and MN assays. Data showed that all five biomaterials did not significantly exert genotoxic effects. Hence, the study indicates that these biomaterials do not induce any chromosomal anomalies.

Antidepressant-like effects of methanol extract of Hibiscus tiliaceus flowers in mice

BackgroundHibiscus tiliaceus L. (Malvaceae) is used in postpartum disorders. Our purpose was to examine the antidepressant, anxiolytic and sedative actions of the methanol extract of H. tiliaceus flowers using animal models.MethodsAdult male Swiss albino mice were treated with saline, standard drugs or methanol extract of H. tiliaceus and then subjected to behavioral tests. The forced swimming and tail suspension tests were used as predictive animal models of antidepressant activity, where the time of immobility was considered. The animals were submitted to the elevated plus-maze and ketamine-induced sleeping time to assess anxiolytic and sedative activities, respectively.ResultsMethanol extract of H. tiliaceus significantly decreased the duration of immobility in both animal models of antidepressant activity, forced swimming and tail suspension tests. This extract did not potentiate the effect of ketamine-induced hypnosis, as determined by the time to onset and duration of sleeping time.ConclusionOur results indicate an antidepressant-like profile of action for the extract of Hibiscus tiliaceus without sedative side effect.

Effect of latex material on antioxidant enzymes, lipid peroxidation, DNA damage, and chromosomal aberration

Cell integrity is affected by oxidative stress when the production of active oxidants overwhelms antioxidant defense mechanisms. Latex, a natural polymer obtained from Hevea brasiliensis, is used in medical industry for manufacturing surgical gloves, urinary catheters, and dental dams. The aim of this study was to evaluate the effects of latex material on oxidative stress by in vivo and in vitro methods. In addition, the material was screened for its ability to induce any chromosomal aberrations (CAs) by in vitro method. In vivo studies were carried out with implanted latex material onto subcutaneous tissue of various batches of experimental Wistar rats. At the end of experimental period, animals were anesthetized, blood was collected for serum analysis, and sacrificed. Liver was excised for the determination of antioxidant enzymes and lipid peroxidation (LPO). Subcutaneous tissues were obtained for the extraction of genomic DNA from implanted animals and checked for the presence of 8-hydroxy-2-deoxyguanosine (8-OHdG), considered an indicator of DNA damage. Simultaneously, in vitro studies were carried out using fresh liver and subcutaneous tissue obtained from Swiss albino mice treated with physiological saline extract of latex material. For the estimation of both in vitro and in vivo oxidative stress, 10% liver homogenate was assessed for stress indicators like reduced glutathione, glutathione reductase, glutathione peroxidase, LPO and protein content. The results of both in vivo and in vitro studies indicated that the chemical leachents from the latex material did not significantly affect LPO and the levels of antioxidant enzymes. There was also no significant increase in 8-OHdG content due to the presence of implanted latex material. Finally, the results of in vitro CA test and G banding indicated that extracts of test material did not induce any chromosomal abnormalities.

Potent activation of tumor necrosis factor-α production by a polysaccharide found in a marine brown alga Chordaria flagelliformis

In a search for new antitumor drugs, we collected 334 species of marine algae from the Japanese coastline and screened these for tumor necrosis factor-alpha (TNF-α) production-promoting activity in an in vitro assay with the mouse macrophage-derived cell line RAW264.7. A phosphate-buffered saline extract from the marine brown alga Chordaria flagelliformis was found to strongly promote TNF-α activity. Further purification of this alga by precipitation with ethanol and two-step chromatography yielded an approximately 3,000-kDa polysaccharide as the active substance, which we designated MC25. MC25 exhibited TNF-α production-promoting activity in RAW264.7 cells in a dose-dependent manner, and we established its temperature and pH stabilities and non-cytotoxic character. Its molecular structure is similar to that of a fucoidan in terms of composition, including glucose, fucose, xylose, and uronic acid. The molecule hydrolyzed to a size of 180–370 kDa still retained strong TNF-α production-promoting activity, suggesting that MC25 might be a promising candidate in the development of novel antitumor agents.

Antioxidant Activity of Moringa oleifera Tissue Extracts

Moringa oleifera is an important source of antioxidants, tools in nutritional biochemistry that could be beneficial for human health; the leaves and flowers are used by the population with great nutritional importance. This work investigates the antioxidant activity of M. oleifera ethanolic (E1) and saline (E2) extracts from flowers (a), inflorescence rachis (b), seeds (c), leaf tissue (d), leaf rachis (e) and fundamental tissues of stem (f). The radical scavenging capacity (RSC) of extracts was determined using dot-blots on thin layer chromatography stained with a 0.4 mM 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) solution; spectrophotometric assays were recorded (515 nm). Antioxidant components were detected in all E1 and E2 from a, b and d. The best RSC was obtained with E1d; the antioxidants present in E2 reacted very slowly with DPPH. The chromatogram revealed by diphenylborinate-2-ethylamine methanolic solution showed that the ethanolic extract from the flowers, inflorescence rachis, fundamental tissue of stem and leaf tissue contained at least three flavonoids; the saline extract from the flowers and leaf tissue revealed at least two flavonoids. In conclusion, M. oleifera ethanolic and saline extracts contain antioxidants that support the use of the plant tissues as food sources.

Thrombocytopenia in a nutshell

In January, 2009, a 70 year old man was referred to us because a blood count done after a night of nausea, vomiting, and fever showed a platelet count of 32×109/L. He had no previous history of thrombocytopenia or haemorrhagic diathesis and did not take medications or quinine-containing beverages. 4 days later, the platelet count was normal, and we believed that the low platelet count might be a result of laboratory error. 8 months later, his symptoms recurred and, again, the platelet count (37×109/L) was low. Other blood values were normal and his platelet count returned to normal in a few days. At this time, our patient recalled eating walnuts before each thrombocytopenic episode, suggesting a possible connection between his complaints and the consumption of walnuts. To test this hypothesis, an in-hospital walnut challenge was undertaken. Our patient’s initial platelet count was 233×109/L. He ate 100 g of walnuts at 1700 h and after 4 h developed a fever (temperature 38·7°C), nausea, and vomiting. No other physical or systemic symptoms, such as rash, angio-oedema, or hypotension, occurred. The nausea and fever resolved during the next 8 h, but he bled from an existing small wound and developed large haematomas at venepuncture sites. His platelet count, measured 15 h after walnut ingestion, was 4×109/L; haemoglobin concentration and white cell count were normal. IgE antibodies specific for tree nuts were not detected. As in previous episodes of thrombocytopenia, the platelet count became normal within 4 days (figure). Figure Changes in platelet count after exposure to walnuts The rapid improvement in platelet count after each episode suggested that the thrombocytopenia might have been caused by a platelet-reactive antibody specific for a substance present in walnuts. We studied new blood samples collected when the platelet count was normal. Strong (titre 1 to 32) IgG antibodies were detected in our patient’s serum by flow cytometry;1 they reacted with platelets when a saline extract made from ground English walnuts(Juglans regia) was present. Similar activity was not detected in normal control serum samples. His antibody did not react with platelets from a patient with Glanzmann’s thrombasthenia lacking the platelet surface αIIb/βIIIa integrin. The findings provided strong evidence that our patient’s thrombocytopenic episodes were caused by an IgG antibody that reacted with αIIb/βIIIa integrin when a soluble substance from walnuts was present. He excluded nuts from his diet and at last follow-up in May 2010, he had had no further symptoms. Many medications are known to cause immune thrombocytopenia, but convincing reports of this complication triggered by foods and beverages are rare. Examples reported were associated with consumption of an African bean,2 sesame seeds,3 cranberry juice,4 cow’s milk, and Jui (a Chinese herbal medicine).5 To the best of our knowledge this is the first case in which a patient reported systemic symptoms and in which a platelet-reactive antibody specific for a substance present in the implicated food was identified. With these findings and the positive provocation test, our patient’s case met all criteria for establishing a causal association between exposure and thrombocytopenia:5 exposure to walnuts resulted in acute thrombocytopenia that rapidly recovered to normal; and no other cause for thrombocytopenia was identified. The provocation test led to profound thrombocytopenia, systemic symptoms, and bleeding, albeit followed by rapid recovery. In retrospect, we should have begun the challenge with a smaller dose of walnuts. Our patient’s case shows the clinical importance of product induced thrombocytopenia and its distinction from other more common causes.

English

Vincristine-induced peripheral neuropathy is a major dose limiting side effect and thus, effective therapeutic strategy is required. In this study, the antinociceptive effect of&nbsp;Matricaria chamomilla&nbsp;(MC) hydroalcoholic extract and morphine on vincristine-induced peripheral neuropathy model in mice has been investigated. Experiments were performed on 60 Naval Medical Research Institute (NMRI) male mice. Mouse subsequently received daily intraperitoneal and intravenal injections of vincristine sulfate, saline and MC hydroalcoholic extract over 12 days, immediately following behavioral testing. For assessment of pain, formalin test was preformed. The effects of MC, morphine and vehicle (saline) 30 min before formalin test on vincristine-induced neuropathy were evaluated. Administration of MC before formalin injection showed significant (P &lt; 0.05) decrease of pain responses in both phases. Administration of vincristine produced significant (Pm &lt; 0.05) increase in pain response in second phase of formalin test. Injection of MC and vincristine together has shown that MC is able to decrease the vincristine induced pain significantly (P &lt; 0.05). Morphine decreased vincristine induced pain test significantly (P &lt; 0.05). In comparison, morphine has analgesic effects in the first phase and MC has anti-inflammatory effects in the second phase of formalin test significantly (P &lt; 0.05). These results suggest that MC may be an alternative approach for the treatment of vincristine-induced peripheral neuropathic pain. &nbsp; Key words:&nbsp;Matricaria chamomilla, vincristine, antinociception,&nbsp;neuropathic pain.

The hepatic protective mechanism of Ginkgo biloba extract in rats with obstructive jaundice

The objective of our study was to examine the hepatic protective mechanism of Ginkgo biloba extract (GBE) in rats with obstructive jaundice (OJ). Twenty rats underwent bile duct ligation and received daily intraperitoneal injections of either control saline or Ginkgo biloba extract for 14 days. Ten sham-operated rats had their bile duct exposed but not ligated or sectioned. Serum alanine transaminase (ALT) was analyzed for liver function tests and liver damage was further assessed by histologic examination. The levels of endothelin 1 (ET-1) and nitric oxide (NO) in blood and liver homogenate were measured. The serum alanine transaminase was elevated in the bile duct ligation rats (BDL rats); GBE could significantly lower serum transaminase level and ameliorate liver histological damage. ET-1 and NO levels in both plasma and liver tissue were also elevated in common bile duct (CBD)-ligated rats, but this increase was significantly decreased by GBE treatment. Moreover, the degree of liver damage severity positively correlates with high levels of ET-1 and NO. GBE mediated the liver protective effect at least in part by suppressing overproduction of ET-1 and NO and restoring a proper balance between ET-1 and NO to some extent.

Involvement of cholinergic-muscarinic receptor in &lt;i&gt;Anaphe venata&lt;/i&gt;-induced stretching-ataxia behavioral effects in rats

Anaphe venata entomophagy has previously been implicated in the aetiopathogenesis of seasonal ataxia in humans and altered motor function in rodents. Thus, we investigated the effect of A. venata PhosphateBuffer Saline (PBS) extract on stretching, ataxia and the possible mechanism(s) of action. Animals were divided into four groups (n = 6-12 per group) and graded doses of extract (100, 200 or 400 mg/kg) wereadministered intraperitoneally (i.p.) while the control group received saline. Behavioral scores were recorded for a period of 30 min immediately after the administration of saline or extract. The role of various receptors inthe extract induced stretching and ataxia was evaluated using known receptor antagonists in other groups of rats. The in-vitro cholinesterase inhibition assay of the extract was also performed. The protein profile of theextract was evaluated using the Sodium Dodecyl Sulphate&nbsp; (SDS)-Polyacrylamide gel electrophoresis. Results showed that the extract induced significant (p&lt;0.01) stretching and ataxia behavioural effects dose-dependently when compared to vehicle-treated rats. Pretreatment with the non-selective muscarinic antagonist, scopolamine, significantly (p&lt;0.05) reversed both stretching and ataxia-induced behaviour of PBS extract at alldose levels however both flumazenil and naloxone did not show any significant effects. Anticholinesterase assay also provided evidence that the extract has inhibitory effect on acetylcholinesterase enzyme. Electrophoresis assay suggested that the major proteins in the extract are probably of small molecular weight. In conclusion, the A. venata PBS extract induced-behaviours are probably mediated via the activation of cholinergic-muscarinic receptor systems.

Assessment of oxidative stress and chromosomal aberration inducing potential of three medical grade silicone polymer materials

Medical expenditures for devices are increasing along with the ageing of human population and the synthesis of materials such as silicone polymers is on upsurge for manufacturing these devices. The International Organization for Standardization (ISO) emphasizes a battery of tests for preclinical assessment of biocompatibility of medical devices. Genotoxicity assays have become an integral component of these test procedures and it employs a set of in vitro and in vivo experiments to detect mutagens. Hence, this study was performed with an intention to investigate the genotoxic potential of the physiological saline extracts of three medical grade silicone polymer materials by the in vitro chromosomal aberration assay using human peripheral blood lymphocytes. Further, the oxidative stress inducing potential of the material extracts was investigated in vivo in mice liver homogenates using cyclophosphamide as positive control. The investigation revealed that none of the three materials were able to produce marked human lymphocyte chromosomal aberration, suggesting the absence of mutagens. The materials also showed negative results in their oxidative stress inducing potential, which was revealed by the normal levels of lipid peroxidation and unaltered levels of glutathione and its metabolizing enzymes in the mice liver tissue homogenates. It was interesting to observe a significant correlation between the genotoxic and antioxidant parameters investigated. Hence, it is suggested that the estimation of antioxidant status would serve as a better preliminary testing procedure prior to evaluating the genetic and molecular toxicity mechanisms of medical devices and/or materials intended for manufacture of such devices.

Immunochemical characterization of antigens of Brucella canis and their use in seroprevalence study of canine brucellosis

To explore immunochemical characterization of antigens of Brucella canis (B. canis), and the use in seroprevalence study of canine brucellosis. External hot phosphate buffer saline extract (HPBSE) and internal sonicated (SA) antigens were prepared from B. canis strain MEX 51 and immunochemically characterized. These antigens were used to test 527 serum samples of dogs by 2-mercaptoethanol-tube agglutination test (2 ME-TAT), agar gel immunodiffusion test (AGID), dot-ELISA and indirect enzyme-linked immunosorbent assay (I-ELISA) to assess the seroprevalence of canine brucellosis. The protein content of HPBSE and SA antigens was 0.387 mg/mL and 0.195 mg/mL, respectively, whereas carbohydrate content was 0.174 mg/mL and 0.150 mg/mL, respectively. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (12.5%) of HPBSE and SA, revealed 6 and 8 visible peptide bands ranging from 18-80 kDa and 12-45 kDa, respectively. Western blot analysis showed immunodominant bands of MW 12, 28, 39 and 45 kDa for HPBSE and 20-24 kDa for SA. The AGID revealed HPBSE as more specific antigen than SA but both I-ELISA and dot-ELISA indicated SA antigen to be more specific and reliable than HPBSE. The seroprevalence of canine brucellosis was 2.27% by 2ME-TAT, 1.5% by AGID, 3.03% by dot-ELISA and 16.12% by I-ELISA. On the basis of the results of present study, we concluded that HPBSE is suitable antigen for AGID, which is more specific; whereas SA antigen is suitable for I-ELISA, which is highly sensitive. Therefore, initial screening of serum samples should be carried out by I-ELISA followed by confirmation with AGID.

Faecal IgA concentration is influenced by age in dogs

Data comparing age-related alterations in faecal IgA concentrations of dogs are not available in the literature. The present study aimed to compare the faecal concentrations of IgA in puppies, mature and senior dogs. A total of twenty-four beagle dogs were used, including eight puppies (5 months old, four females and four males), eight mature (4.6 years old, eight males) and eight senior dogs (10.6 years old, three males and five females). Fresh faecal samples were collected from each dog for three consecutive days and pooled by animal. After saline extraction, IgA content was measured by ELISA. Data were analysed by one-way ANOVA, and means were compared with Tukey's test (P < 0.05). Results showed that puppies have lower faecal IgA concentrations than mature dogs (P < 0.05); senior animals presented intermediary results. The reduced faecal IgA concentration in puppies is consistent with the reduced serum and salivary IgA concentrations reported previously, suggesting a reduced mucosal immunity in this age group. Although some studies have found an increased serum IgA concentration in older dogs, this may differ from the intestinal secretion of IgA, which appears to be lower in some senior animals (four of the eight dogs studied).

Delivery of nicotine in an extract of a smokeless tobacco product reduces its reinforcement-attenuating and discriminative stimulus effects in rats

Animal models of tobacco addiction rely on administration of nicotine alone or nicotine combined with isolated constituents. Models using tobacco extracts derived from tobacco products and containing a range of tobacco constituents might more accurately simulate tobacco exposure in humans. To compare the effects of nicotine alone and an aqueous smokeless tobacco extract in several addiction-related animal behavioral models. Nicotine alone and nicotine dose-equivalent concentrations of extract were compared in terms of their acute effects on intracranial self-stimulation (ICSS) thresholds, discriminative stimulus effects, and effects on locomotor activity. Similar levels of nicotine and minor alkaloids were achieved using either artificial saliva or saline for extraction, supporting the clinical relevance of the saline extracts used in these studies. Extract produced reinforcement-enhancing (ICSS threshold-decreasing) effects similar to those of nicotine alone at low to moderate nicotine doses, but reduced reinforcement-attenuating (ICSS threshold-increasing) effects at a high nicotine dose. In rats trained to discriminate nicotine alone from saline, intermediate extract doses did not substitute for the training dose as well as nicotine alone. Locomotor stimulant effects and nicotine distribution to brain were similar following administration of extract or nicotine alone. The reinforcement-attenuating and discriminative stimulus effects of nicotine delivered in an extract of a commercial smokeless tobacco product differed from those of nicotine alone. Extracts of tobacco products may be useful for evaluating the abuse liability of those products and understanding the role of non-nicotine constituents in tobacco addiction.

Extraction and mass spectrometry identification of a major peach allergen Pru p 1

Peach allergy can be caused by the allergen Pru p 1. This occurs by cross-reactivity with the homologous birch pollen allergen Bet v 1. However, the direct identification of Pru p 1 as an immunoglobulin E (IgE)-binding protein extracted from peach fruit has never been reported. Phosphate-buffered saline (PBS) and phenol extractions were applied to solubilise the proteins from peach peel and pulp, and IgE immunoblotting with sera of individual peach-allergic patients was used to detect the potential allergens. Most of the patients showed binding to an 18 kDa band in IgE immunoblotting performed with the phenolic extracts of peach peel and pulp, but not when the PBS extracts were used. Mass spectrometry of the 18 kDa spot excised from a two-dimensional electrophoretic gel showed this protein to correspond to the peach allergen Pru p 1. Phenol extraction was necessary to detect by IgE immunoblotting a major peach allergen, which showed very low extractability with PBS, indicating the appropriateness of adopting different extraction procedures to identify plant allergens. The 18 kDa peach protein was definitively identified as the Bet v 1-homologous peach allergen Pru p 1.

Evaluation of hydroxyapatite–bioglass and hydroxyapatite–ethyl vinyl acetate composite extracts on antioxidant defense mechanism and genotoxicity: An in vitro study

Hydroxyapatite–bioglass (HA BG) and hydroxyapatite–ethyl vinyl acetate (HA EVA) are two composite materials that have been developed for bone substitution. Their activity on antioxidant defense mechanism and genotoxicity has not been investigated before. To further confirm its biocompatibility, the present study was undertaken to investigate the effect of HA BG and HA EVA on mice liver antioxidant mechanism along with chromosomal aberrations in human lymphocytes. Physiological saline extract of HA BG and HA EVA showed no adverse effect on liver antioxidant mechanism compared to the cyclophosphamide (CP)-induced toxicity on mice liver homogenate. The results were judged from the in vitro studies made on reduced glutathione, glutathione reductase and lipid peroxidation. These results were well supported by CP- and mytomycin C (MC)-induced genotoxicity studies on human lymphocytes in the presence and absence of a metabolic activator (S9). Hence, it was suggested that these tests could be considered for preliminary toxicological screening of materials intended for clinical applications ahead of in vivo animal model evaluation.