Development of a Low Cost Semiquantitative Polymerase Chain Reaction Assay for Molecular Diagnosis of Williams Syndrome.

BACKGROUND: Williams Beuren Syndrome (WBS) is a well-recognized and common genetic cause of congenital heart defects, developmental delay, hypercalcemia, and characteristic facial features. It is caused by a 1.5 - 1.8 Mb heterozygous deletion of chromosome 7q11.23 with loss of around 28 coding genes. The aim of this study was to develop a low-cost, semi-quantitative PCR (sqPCR) method to detect the chromosome 7q11.23 deletion. METHODS: Twenty-four suspected WBS cases were recruited following ethical clearance and informed consent. Blood was obtained, DNA extracted and spectrophotometrically quantified using standard methods. To detect the deletion by dosage analysis, a target region within a gene located in the WBS commonly deleted region of 7q11.23 was amplified together with a control region in a duplex sqPCR assay. The control region was telomeric to the WBS commonly deleted region and was located in chromosome 7q31.2. The two target regions within the deleted region namely a locus within ELN and a marker in the intergenic region between FZD9 and FKBP6 and designated IFF, were amplified in separate duplex sqPCR assays. The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene was used as the control for normalization. Included in the assay were a non-deleted and deleted individuals' samples. RESULTS: Nineteen patients were identified to have the deletion while five did not. All 24 patients' results were confirmed by whole exome sequencing and 11 also by fluorescence in-situ hybridization (FISH). CONCLUSIONS: The data obtained indicates the sqPCR assay developed in this study to be an accurate and reliable diagnostic test for WBS. Most Sri Lankan patients with WBS are diagnosed clinically, as many parents of affected WBS children are unable to afford currently available molecular diagnostic testing. This low cost sqPCR test is therefore likely to benefit Sri Lankan WBS patients, by enabling genetic testing for confirming or refuting a clinical diagnosis of WBS and may be of use in other low and middle income countries.

Cannabinoid type 2 receptor inhibition enhances the antidepressant and proneurogenic effects of physical exercise after chronic stress.

Chronic stress is a major risk factor for neuropsychiatric conditions such as depression. Adult hippocampal neurogenesis (AHN) has emerged as a promising target to counteract stress-related disorders given the ability of newborn neurons to facilitate endogenous plasticity. Recent data sheds light on the interaction between cannabinoids and neurotrophic factors underlying the regulation of AHN, with important effects on cognitive plasticity and emotional flexibility. Since physical exercise (PE) is known to enhance neurotrophic factor levels, we hypothesised that PE could engage with cannabinoids to influence AHN and that this would result in beneficial effects under stressful conditions. We therefore investigated the actions of modulating cannabinoid type 2 receptors (CB2R), which are devoid of psychotropic effects, in combination with PE in chronically stressed animals. We found that CB2R inhibition, but not CB2R activation, in combination with PE significantly ameliorated stress-evoked emotional changes and cognitive deficits. Importantly, this combined strategy critically shaped stress-induced changes in AHN dynamics, leading to a significant increase in the rates of cell proliferation and differentiation of newborn neurons, overall reduction in neuroinflammation, and increased hippocampal levels of BDNF. Together, these results show that CB2Rs are crucial regulators of the beneficial effects of PE in countering the effects of chronic stress. Our work emphasises the importance of understanding the mechanisms behind the actions of cannabinoids and PE and provides a framework for future therapeutic strategies to treat stress-related disorders that capitalise on lifestyle interventions complemented with endocannabinoid pharmacomodulation.

[Macrovipera lebetina obtusa Snake Venom as a Modulator of Antitumor Effect in S-180 Sarcoma Mouse Model].

Macrovipera lebetina obtusa (MLO) is a venomous snake endemic to Middle East. Here we describe the therapeutic potential of the MLO snake venom. In S-180 sarcoma-bearing mouse model, we showed that the MLO snake venom inhibits tumour growth by 50%. In human dermal microvascular endothelial cells (HMVEC-D), treatment with the MLO snake venom lead to an increase of expression levels of the vascular endothelial growth factor (VEGF), while the level of the expression of caspase 8 did not change. In HMVEC-D cells MLO snake venom induces necroptosis, rather than apoptosis. In the chick embryo chorioallantoic membrane (CAM) assay, exposure to MLO snake venom inhibited bFGF-induced angiogenesis by 22%. Taken together, these results indicate that the MLO snake venom has a potent cytotoxic activity. Regulated necroptic cell death pathway, which is engaged by MLO snake venom, may become a promising novel target for antitumor therapies.

Deep Brain Stimulation of the dorsal raphe abolishes serotonin 1A facilitation of AMPA receptor-mediated synaptic currents in the ventral hippocampus.

In a previous study we showed that Deep Brain Stimulation (DBS) of the rat dorsal subregion of the dorsal raphe (DRD), which sends serotonergic projections to forebrain areas, such as the ventral hippocampus, induces anxiolytic-like effects. The purpose of the present study was to investigate neurobiological alterations which might underline these behavioral effects. For that, we tested the influence of DBS upon the neuromodulatory action of serotonin on excitatory post-synaptic currents (EPSCs) in the ventral hippocampus. Male Wistar rats were submitted to high-frequency stimulation (100 µA, 100 Hz) of the DRD for 1 h during three consecutive days. On the third day, immediately after the DBS procedure, animals were euthanized. Slices of the ventral hippocampus were processed for whole cell patch clamp recordings of AMPA-receptor (AMPAR) mediated EPSCs in the CA1 area. As reported by others, we confirmed that in pre-weaning rats a high affinity 5-HT1A receptor agonist (8-OH-PIPAT, 0.5-5nM) inhibits EPSCs. However, in adult rats (non-operated or sham-operated), 8-OH-PIPAT (0.5-5 nM) increased EPSC amplitude, an effect blocked by the 5-HT1A antagonist WAY-100,635 (200 nM). Importantly, in adult rats exposed to DBS, the 5-HT1A agonist was devoid of effect. Taken together these results show that: 1) changes in 5-HT1A receptor-mediated hippocampal synaptic transmission occur with age; 2) these changes lead to a facilitatory effect of 5-HT1A receptors; 3) DBS blocks this serotonergic facilitatory action. These observations suggest that an alteration in serotonin modulation of limbic areas may underlie the psychotherapeutic effects of DBS.

Hippocampal synaptic dysfunction in the SOD1G93A mouse model of Amyotrophic Lateral Sclerosis: Reversal by adenosine A2AR blockade.

Amyotrophic Lateral Sclerosis (ALS) mostly affects motor neurons, but non-motor neural and cognitive alterations have been reported in ALS mouse models and patients. Here, we evaluated if time-dependent biphasic changes in synaptic transmission and plasticity occur in hippocampal synapses of ALS SOD1G93A mice. Recordings were performed in hippocampal slices of SOD1G93A and age-matched WT mice, in the pre-symptomatic and symptomatic stages. We found an enhancement of pre-synaptic function and increased adenosine A2A receptor levels in the hippocampus of pre-symptomatic mice. In contrast, in symptomatic mice, there was an impairment of long-term potentiation (LTP) and a decrease in NMDA receptor-mediated synaptic currents, with A2AR levels also being increased. Chronic treatment with the A2AR antagonist KW-6002, rescued LTP and A2AR values. Altogether, these findings suggest an increase in synaptic function during the pre-symptomatic stage, followed by a decrease in synaptic plasticity in the symptomatic stage, which involves over-activation of A2AR from early disease stages.

Evaluation of the biocompatibility and skin hydration potential of vitamin E-loaded lipid nanosystems formulations: In vitro and human in vivo studies.

Lipid-based nanosystems, such as nanostructured lipid carriers (NLC) and nanoemulsions (NE) have been described as promising alternatives to conventional formulations for increase skin hydration. Besides, these systems have been used as efficient vehicles for lipophilic molecules that improve skin properties (e.g. vitamin E). In this study, we performed comparative investigations between hydrogels formulations containing vitamin E-loaded NLC (HG-NLCVE) and vitamin E-loaded nanoemulsion (HG-NEVE). The experiments started with particle size measurements, which showed no significant differences between nanoparticles/nanodroplets sizes after incorporation in the hydrogel net (386 nm vs. 397 nm for HG-NLCVE and 402 nm vs. 514 nm for HG-NEVE). Afterwards, in vitro biocompatibility studies in human keratinocytes were carried out, being observed that the lipid-based nanosystems were more cytotoxic for the cells before incorporation in the hydrogel. Finally, the formulations hydration potential and sensory attributes for skin application were evaluated by in vitro occlusion tests and in vivo human experiments. The results showed that the HG-NLCVE exhibited the best occlusive properties, whereas the HG-NEVE performed a faster skin hydration effect. Furthermore, the latter was selected as the most attractive for skin application, although the HG-NLCVE was described as more suitable to obtain a long-lasting effect. This study demonstrated the in vitro and in vivo safety and hydration potential of hydrogels containing vitamin E-loaded lipid-based nanosystems. These results establish a basis to assess the cutaneous use of these systems, despite more in vivo experiments, for longer periods and in more volunteers, are required before commercialization.

Detection of bacterial pathogens from clinical specimens using conventional microbial culture and 16S metagenomics: a comparative study.

BACKGROUND: Infectious disease is the leading cause of death worldwide, and diagnosis of polymicrobial and fungal infections is increasingly challenging in the clinical setting. Conventionally, molecular detection is still the best method of species identification in clinical samples. However, the limitations of Sanger sequencing make diagnosis of polymicrobial infections one of the biggest hurdles in treatment. The development of massively parallel sequencing or next generation sequencing (NGS) has revolutionized the field of metagenomics, with wide application of the technology in identification of microbial communities in environmental sources, human gut and others. However, to date there has been no commercial application of this technology in infectious disease diagnostic settings. METHODS: Credence Genomics Rapid Infection Detection™ test, is a molecular based diagnostic test that uses next generation sequencing of bacterial 16S rRNA gene and fungal ITS1 gene region to provide accurate identification of species within a clinical sample. Here we present a study comparing 16S and ITS1 metagenomic identification against conventional culture for clinical samples. Using culture results as gold standard, a comparison was conducted using patient specimens from a clinical microbiology lab. RESULTS: Metagenomics based results show a 91.8% concordance rate for culture positive specimens and 52.8% concordance rate with culture negative samples. 10.3% of specimens were also positive for fungal species which was not investigated by culture. Specificity and sensitivity for metagenomics analysis is 91.8 and 52.7% respectively. CONCLUSION: 16S based metagenomic identification of bacterial species within a clinical specimen is on par with conventional culture based techniques and when coupled with clinical information can lead to an accurate diagnostic tool for infectious disease diagnosis.

The impact of water pollution on fish species in southeast region of Goiás, Brazil.

The rivers from the region of Catalão, Southeast Goiás State, Brazil, are exposed to intense anthropogenic influences including agricultural activities, industry, and urban waste. The aim of this study was to determine the severity of water pollution by conducting an experiment involving in situ biomonitoring of water constituents on genotoxicity in fish inhabiting these sites. The genotoxicity of three sites of the region were analyzed utilizing the micronucleus (MN) test. It was of interest to determine whether there were differences between sampling sites such as urban perimeter, agriculture, and fertilizer industry in control, and monitored species including Astyanax fasciatus, Astyanax altiparanae, and Characidium fasciatum. Data demonstrated that the species at sites 1, 2, and 3 exhibited a marked increase in frequency of MN compared to fish from site 4. Significant elevation in frequency of MN occurred in erythrocytes of A. fasciatus and A. altiparanae at sites 1 and 2. At site 3 higher frequencies of MN were observed in C. fasciatum. MN induction in C. fasciatum was correlated with chromium levels in water and sediment, while A. fasciatum and A. altiparanae showed an association with zinc in water and sediment. Data suggest that benthic and nektonic fish species display different sensitivities in relation to anthropogenic contaminant influences.

Breast cancer lymphoscintigraphy: Factors associated with sentinel lymph node non visualization.

OBJECTIVE: To evaluate factors associated with non identification of the sentinel lymph node (SLN) in lymphoscintigraphy of breast cancer patients and analyze the relationship with SLN metastases. MATERIAL AND METHODS: A single-center, cross-sectional and retrospective study was performed. Forty patients with lymphoscintigraphy without sentinel lymph node identification (negative lymphoscintigraphy - NL) were enrolled. The control group included 184 patients with SLN identification (positive lymphoscintigraphy - PL). Evaluated factors were age, body mass index (BMI), tumor size, histology, localization, preoperative breast lesion hookwire (harpoon) marking and SLN metastases. The statistical analysis was performed with uni- and multivariate logistic regression models and matched-pairs analysis. RESULTS: Age (p=0.036) or having BMI (p=0.047) were the only factors significantly associated with NL. Being ≥60 years with a BMI ≥30 increased the odds of having a NL 2 and 3.8 times, respectively. Marking with hookwire seems to increase the likelihood of NL, but demonstrated statistical significance is lacking (p=0.087). The other tested variables did not affect the examination result. When controlling for age, BMI and marking with the harpoon, a significant association between lymph node metastization and NL was not found (p=0.565). CONCLUSIONS: The most important factors related with non identification of SLN in the patients were age, BMI and marking with hook wire. However, only the first two had statistical importance. When these variables were controlled, no association was found between NL and axillary metastases.

Impact of root fillings and posts on the diagnostic ability of three intra-oral digital radiographic systems in detecting vertical root fractures.

AIM: To evaluate (i) the accuracy of three intra-oral digital systems in detecting vertical root fractures (VRFs); and (ii) the influence of different intracanal conditions (without filling, filled with Gutta-percha and metal or fibreglass post). METHODOLOGY: Forty single-rooted human teeth were decoronated and the root canals prepared. VRFs were induced using a universal testing machine on 20 teeth. Radiographic images were obtained using the paralleling technique with three different horizontal angulations on conventional E/F film, two storage phosphor plates (Digora Optime and VistaScan) and one sensor system (SnapShot). All teeth were radiographed with all intracanal conditions tested. All images were evaluated by five examiners. After 30 days, 25% of the images were reviewed. Areas under ROC curves, sensitivity, specificity, accuracy, positive predictive and negative predictive values were compared by anova. RESULTS: The weighted kappa values for intra- and interobserver reproducibility were 0.48-0.74 and 0.57-0.66, respectively. The sensitivity revealed a significant difference between imaging modalities, with higher values for SnapShot (0.53) and VistaScan (0.46), followed by the film (0.41) and Digora (0.38). The area under the ROC curve for SnapShot (0.82) was greater than for the other systems in the teeth with a fibreglass post (VistaScan: 0.70; Digora: 0.64; film: 0.60). CONCLUSION: All systems gave a similar performance for detecting VRFs; all gave a good performance in the absence of intracanal materials, but performed less well in the presence of a metal post or Gutta-percha. In teeth with a fibreglass post, digital systems with higher resolution should be used.

The 'Sharpen' filter improves the radiographic detection of vertical root fractures.

AIM: To compare the diagnostic accuracy of digital enhancement filters in the diagnosis of vertical root fractures. METHODOLOGY: The sample consisted of 40 single-rooted teeth randomly divided into two groups: experimental and control. Vertical root fractures were induced in the experimental group using a universal testing machine. All teeth were individually radiographed with three different horizontal angles using the Digora Optime(®) digital system. Three observers separately examined the original and filtered images (3D Emboss, Negative, Sharpen and Shadow). The area under Receiver Operating Characteristic curve values (Az) for each protocol were compared by one-way anova with post hoc Bonferroni test. The significance level was set at 5%. RESULTS: The highest Az value was obtained using the Sharpen filter, with significant differences from the original and other filtered images (P < 0.05). The 3D Emboss filter had the lowest Az value, which was significantly different from the other filtered images (P < 0.05). CONCLUSION: When using the Digora Optime(®) system, the 'Sharpen' filter is recommended to improve the radiographic detection of vertical root fractures.

Morphological variability of the shape of striped red mullet Mullus surmuletus in relation to stock discrimination between the Bay of Biscay and the eastern English Channel.

Truss analysis and length measurements were made on 168 striped red mullet Mullus surmuletus. Multivariate statistical analyses with principal component analysis and partial redundancy analysis (pRDA) were used on these measurements to evaluate the influence of maturity, sex and geographical area distribution on body shape. Truss measurements were important to quantify and discriminate changing body shape, presumably due to changing environmental conditions. Sexual dimorphism was not observed and juveniles could be distinguished from adults based on their body shape. More importantly, M. surmuletus occurring in different geographical areas could be differentiated using this method. Based on pRDA, a significant difference of head morphological dimensions was observed between populations occurring in the eastern English Channel and those occurring in the Bay of Biscay, suggesting that fish from these areas could represent two subpopulations.

Antifungal activity of Ferulago capillaris essential oil against Candida, Cryptococcus, Aspergillus and dermatophyte species.

This study evaluates the composition, antifungal activity and mechanism of action of the essential oil of Ferulago capillaris (Link ex Spreng.) Cout. and its main components, limonene and α-pinene, against clinically relevant yeasts and moulds. Essential oil from the plant's aerial parts was obtained by hydrodistillation and analysed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC-MS). Essential oil showed high contents of limonene (30.9%) and α-pinene (35.8%). Minimum inhibitory concentrations (MICs) were measured according to the reference Clinical and Laboratory Standards Institute (CLSI) broth macrodilution protocols. Cell suspensions were subcultured in solid medium and the minimum fungicidal concentrations (MFCs) were rendered. The effect of essential oil on germ tube formation, mitochondrial function and ergosterol biosynthesis was investigated. Essential oil and α-pinene displayed low and similar MIC and MFC values against tested organisms (0.08 to 5.0 µL/mL), while limonene showed a weaker activity (0.32 to 20 µL/mL). Essential oil inhibited germ tube formation at sub-inhibitory concentrations on Candida albicans. The exposure of C. albicans to the essential oil resulted in impairment of mitochondrial functions in a dose-dependent manner. No difference in ergosterol content was observed in essential oil-treated C. albicans. F. capillaris and α-pinene display a broad fungicidal activity. The fungicidal activity of F. capillaris on C. albicans can be related to an induced oxidative stress which affects enzymes activity and the membrane potential of mitochondria. The essential oil of F. capillaris was shown to have potential for use in the development of clinically useful therapeutic preparations, particularly for topical application in the management of superficial mycoses.

Lavandula luisieri essential oil as a source of antifungal drugs.

This work reports the antifungal activity of Lavandula luisieri essential oils against yeast, dermatophyte and Aspergillus strains responsible for human infections and food contamination. The oil's cytotoxicity and its effect on the yeast-mycelium transition in Candida albicans, an important virulence factor, were also evaluated. Analyses by GC and GC/MS showed a peculiar composition of irregular monoterpenes. Significant differences between the samples occurred in the amounts of 1,8-cineole, fenchone and trans-α-necrodyl acetate. The oil with higher amounts of irregular monoterpenes was the most effective. The influence of the oils on the dimorphic transition in C. albicans was also studied through the germ tube inhibition assay. Filamentation was completely inhibited at concentrations sixteen times lower than the minimal inhibitory concentration. The results support the use of L. luiseiri essential oils in the development of new phytopharmaceuticals and food preservatives and emphasise its antifungal properties at concentrations not cytotoxic or with very low detrimental effects on mammalian cells.

Intake, digestibility and intake behaviour in cattle fed different levels of palm kernel cake / Consumo, digestibilidad, y comportamiento ingestivo del ganado alimentado con diferentes niveles de torta de palma

Objective. The potential use of palm kernel cake was evaluated as a replacement for soybean and corn meal in cattle feed, by investigating their intake, digestibility levels and the intake behaviour of cattle fed diets containing different levels of palm kernel cake concentrate. Materials and methods. The experiment was conducted at the Experimental Farm of the Federal University of Bahia, between August and October 2009. Five crossbred Holstein x Zebu adults, were used. A 5 x 5 Latin square experimental design was used. The animals were fed Tifton-85 Bermudagrass, which made up 65% of their diet, plus one of five different levels of palm kernel cake concentrate (0, 7, 14, 21 and 28%). Results. A linear decrease in dry matter (kg/day) was observed due to the lower palatability and higher fiber content of the palm kernel cake. Neutral detergent fiber intake by the animals showed a quadratic behavior. The coefficients of fractional digestibilities of the analyzed feed, did not differ due to the inclusion of palm kernel cake. The ingestive behavior of the animals was not influenced by the inclusion of palm kernel cake in the diet. Conclusions. Palm kernel cake can be used as an alternative feed supplement in ruminant production systems to reduce feed costs without changes in the studied variables.

Objetivo. Se evaluó el uso potencial de la torta de palma como sustituto de harina de soja y harina de maíz en la alimentación del ganado mediante el estudio de su consumo, niveles de digestibilidad y el comportamiento ingestivo de los bovinos alimentados con dietas con diferentes niveles de torta de palmiste. Materiales y métodos. El experimento se realizó en la Granja Experimental de la Universidad Federal de Bahía, entre agosto y octubre de 2009. Cinco animales cruzados Holstein x Cebú, fueron utilizados. Se utilizó un diseño experimental de cuadrado latino 5x5. Los animales fueron alimentados con heno de Tifton-85 Bermuda (65% de la dieta). Además, cinco niveles diferentes de concentrado de palma torta de palmiste (0, 7, 14, 21 y 28%). Resultados. Se observó una disminución lineal en la materia seca (kg/día) debido a la palatabilidad inferior y al contenido de fibra superior de la torta de palmiste. La ingesta de fibra en detergente neutro por los animales mostró un comportamiento de segundo grado. Los coeficientes de digestibilidad fraccional de los alimentos analizados no fueron diferentes debido a la inclusión de la torta de palma. El comportamiento ingestivo de los animales no fue afectado por la inclusión de hasta 28% de torta de palma en la dieta. Conclusiones. La torta de palma se puede utilizar como un suplemento alimenticio alternativo en los sistemas de producción de rumiantes para reducir los costos de alimentación, sin cambios de las variables estudiadas.

Antifungal activity of phenolic-rich Lavandula multifida L. essential oil.

This study evaluates the antifungal activity and mechanism of action of a new chemotype of Lavandula multifida from Portugal. The essential oil was analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS), and the minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) of the oil and its major compounds were determined against several pathogenic fungi responsible for candidosis, meningitis, dermatophytosis, and aspergillosis. The influence of the oil on the dimorphic transition in Candida albicans was also studied, as well as propidium iodide (PI) and FUN-1 staining of C. albicans cells by flow cytometry. The essential oil was characterized by high contents of monoterpenes, with carvacrol and cis-ß-ocimene being the main constituents. The oil was more effective against dermatophytes and Cryptococcus neoformans, with MIC and MLC values of 0.16 µL/mL and 0.32 µL/mL, respectively. The oil was further shown to completely inhibit filamentation in C. albicans at concentrations below the respective MIC (0.08 µL/mL), with cis-ß-ocimene being the main compound responsible for this inhibition (0.02 µL/mL). The flow cytometry results suggest a mechanism of action ultimately leading to cytoplasmic membrane disruption and cell death. L. multifida essential oil may be useful in complementary therapy to treat disseminated candidosis, since the inhibition of filamentation alone appears to be sufficient to treat this type of infection.

Evaluation of reconstructed images with different voxel sizes of acquisition in the diagnosis of simulated external root resorption using cone beam computed tomography.

AIM: To evaluate the diagnosis of simulated external root resorption in cone beam computed tomography (CBCT) images acquired with different voxel sizes, then reconstructed with the same voxel size. METHODOLOGY: Buccal, mesial and distal root surfaces of twenty single-rooted premolar teeth were evaluated for the presence or absence of simulated resorption defects. Images were obtained using a Classic i-CAT CBCT unit with two different voxel sizes (0.25 and 0.30 mm). Then all images were reconstructed to 0.25 mm voxel size. The McNemar's test was applied to evaluate the agreement between the two protocols (different voxel sizes/reconstruction parameters) with the gold standard (defects created in the root). Accuracy, sensitivity, specificity, positive and negative predictive values were calculated for each voxel and defect sizes. RESULTS: Better sensitivity and accuracy were obtained with 0.25 (0.25 mm) voxel size. Specificity, positive and negative predictive values were highest with the 0.25 (0.30 mm) voxel size. McNemar's test results revealed significant differences between the protocols with the gold standard (P < 0.0001). The results revealed that as the cavity size increased, sensitivity, accuracy, positive and negative predictive values also increased. When correlating the voxel size and root third, better sensitivity, accuracy, positive and negative predictive values were obtained in the middle third. CONCLUSIONS: In the diagnosis of simulated external root resorption with the Classic i-CAT CBCT unit, images acquired with 0.30 mm voxel size and reconstructed with 0.25 mm voxel size provided the best protocol with a lower radiation dose than the 0.25 mm voxel size.

In vivo hydroquinone exposure impairs allergic lung inflammation in rats.

Hydroquinone (HQ) is naturally found in the diet, drugs, as an environmental contaminant and endogenously generated after benzene exposure. Considering that HQ alters the immune system and its several source of exposures in the environment, we hypothesized that prolonged exposure of HQ could affect the course of an immune-mediated inflammatory response. For this purpose, male Wistar rats were intraperitoneally exposed to vehicle or HQ once a day, for 22 days with a 2-day interval every 5 days. On day 10 after exposure with vehicle or HQ, animals were ovalbumin (OA)-sensitized and OA-aerosolized challenged on day 23. HQ exposure did not alter the number of circulating leukocytes but impaired allergic inflammation, evidenced by lower number of leukocytes in the bronchoalveolar lavage fluid 24h after OA-challenge. Reduced force contraction of ex vivo tracheal segments upon OA-challenge and impaired mesentery mast cell degranulation after in situ OA-challenge were also detected in tissues from HQ exposed animals. The OA-specificity on the decreased responses was corroborated by normal trachea contraction and mast cell degranulation in response to compound 48/80. In fact, lower levels of circulating OA-anaphylactic antibodies were found in HQ exposed rats, as assessed by passive cutaneous anaphylaxis assay. The reduced level of OA-anaphylactic antibody was not dependent on lower number or proliferation of lymphocytes. Nevertheless, lower expression of the co-stimulatory molecules CD6 and CD45R on OA-activated lymphocytes from HQ exposed rats indicate the interference of HQ exposure with signaling of the humoral response during allergic inflammation. Together, these data indicate specific effects of HQ exposure manifested during an immune host defense.