Samples Of Different Tissues Research Articles

Immune response of the Almaco jack (Seriola rivoliana) against the infestation with Neobenedenia sp. in three cultivated temperatures.

The Almaco jack (Seriola rivoliana) is a marine fish maintained in mariculture systems and frequently infested by monogenean parasites like Neobenedenia sp. Severe infestations can lead to high mortalities and economic losses for farmers. This study evaluated the effects of temperature on the immune response on Almaco jack infested with Neobenedenia sp. We exposed infested fishes at temperatures of 20°C, 24°C, and 30°C for 20days and took samples of different tissues at the beginning of the experiment, and after 3 and 20days. The tissues considered were the skin, thymus, cephalic kidney, and spleen to evaluate the relative gene expression of different genes: Hsp70, IgM, IL-1β, IL-10, and MyD88. Our results showed an increase in IL-1β gene expression in the skin after 20days of infestation but no significant effect of temperature on gene expression, despite increases in infestation rates with temperature. Therefore, relative genetic expression was controlled by the number of parasites and the days post-infestation. These results show that the parasite infestation induced a local response in the skin, but that temperature has an indirect effect on the immune system of Almaco jack.

Potential of Suaeda nudiflora and Suaeda fruticosa to Adapt to High Salinity Conditions

The deposition of salts in soil seems likely to become a significant barrier for plant development and growth. Halophytes that flourish in naturally saline habitats may sustain extreme salt levels by adopting different acclimatory traits. Insight into such acclimatory features can be useful for devising salt-resilient crops and the reclamation of saline soil. Therefore, salinity-induced responses were studied in two halophytes, i.e., Suaeda nudiflora and Suaeda fruticosa, at a high soil salinity level (ECe 65) to explore their possible tolerance mechanisms in their natural habitat. Samples of different tissues were collected from both Suaeda species for the determination of physio-biochemical attributes, i.e., ionic (Na+, K+, Ca2+, Cl−) content, osmo-protective compounds (proline, soluble sugars, soluble proteins), total phenolic content, and antioxidant components. Heavy metal composition and accumulation in soil and plant samples were also assessed, respectively. Fourier transform infrared spectroscopy (FTIR) analysis was conducted to explore cellular metabolite pools with respect to high salinity. The results showed that both species considerably adjusted the above-mentioned physio-biochemical attributes to resist high salinity, demonstrated by quantitative differences in their above-ground tissues. The FTIR profiles confirmed the plants’ differential responses in terms of variability in lipids, proteins, carbohydrates, and cell wall constituents. The high capacity for Na+ and Cl− accumulation and considerable bioaccumulation factor (BAF) values for metals, mainly Fe and Zn, validate the importance of both Suaeda species as phytodesalination plants and their potential use in the phytoremediation of salt- and metal-polluted soils.

Histological Studies of Snails at Different Periods of Their Life Activity

Large species of terrestrial mollusks are used for human consumption and are bred for this purpose. They have become a part of national cuisine in many countries. Histology is used in many fields of humane and veterinary medicine. Its main essence is that this method can diagnose the presence of formations and inclusions. It is used to determine this process's structure and stage of development. Samples of different tissues can be used as material for histological examination. The specialist analyzes and describes the histological preparations obtained after processing the material. Thus, we proposed a histological method of studying snails. This method can provide the data of snails’ state as a whole. Snails of the species Helix aspersa maxima, Helix aspersa muller, and Helix pomatia were the material for the study. Ten snails of each species were selected for the experiment at different periods: immediately after hibernation, five days after wakening, 14 days after wakening, and at the time of their active life – in July. First, we conducted histological studies of snails during their active life, i.e., in the summer months. Therefore, for this purpose, snails for the experiment were taken in July. It was determined that if the snail eats and has an everyday life, its digestive gland generally consists of digestive tubules separated by intertubular connective tissue containing hemolymphatic sinuses and hemocytes. Thus, in our research, we studied the structure of the digestive gland. A circular muscle layer surrounds each tubule. There are three different cell types in the epithelium lining the tubules of the digestive glands, which differentiate into digestive cells, calcium cells, and excretory cells. Digestive cells are the most numerous cellular component of the tubular epithelium of the digestive gland. Basally located nuclei of digestive cells have a round or oval shape. During the rest period, the digestive gland of snails is characterized by a complete loss of digestive function and the transformation of digestive cells into excretory ones. Metabolic products are not excreted into the environment. When they accumulate, they fill all cavities in the body of snails, namely the lumens of glandular tubules, excretory ducts, stomach, and intestines. Food intake in snails increases the level of metabolism. It stimulates the differentiation of digestive cells, the functional activity of excretory cells, and the volume of excretory products. It leads to the complete liberation of the digestive tract from accumulated excretion products during the rest period. The structure of the digestive gland is completely restored.

Evaluation of Reference Genes for Gene Expression Analysis in Eichhornia crassipes

Eichhornia crassipes is a notorious invasive aquatic weed, causing enormous ecological and economic losses worldwide. However, it has great potential in agriculture, industry, medical care, and other areas. While being such an important plant, it is poorly understood from the molecular perspective. Aiming to select suitable reference genes for gene expression quantification in E. crassipes, this study favors future research at the molecular level. In this work, 12 candidate reference genes were selected. Their expression stability in samples of different tissues, samples treated with various hormones, samples supplied with different levels of phosphorus (P), and pooled samples, were analyzed using GeNorm, NormFinder, BestKeeper, and RefFinder. Meanwhile, the optimal number of reference genes was calculated by GeNorm. The results showed that eIF and ElF1a were the two most stable reference genes in all samples and in tissue samples. In response to hormone treatments, Actin and eIF are the best choices of internal controls. In the case of P treatments, TUA and H2A are recommended to be used as reference genes. Overall, results from this work suggest different reference genes should be applied in qRT-PCR on E. crassipes, according to the specific experimental setup.

Dietary High Dose of Iron Aggravates the Intestinal Injury but Promotes Intestinal Regeneration by Regulating Intestinal Stem Cells Activity in Adult Mice With Dextran Sodium Sulfate-Induced Colitis.

The effects of excessive dietary iron intake on the body have been an important topic. The purpose of this study was to investigate the effects of high-dose iron on intestinal damage and regeneration in dextran sodium sulfate (DSS)-induced colitis model mice. A total of 72 8-week-old adult C57BL/6 mice were randomly divided into two dietary treatment groups: the basal diet supplemented with 45 (control) and 450 mg/kg iron (high-iron) from ferrous sulfate. The mice were fed different diets for 2 weeks, and then 2.5% DSS was orally administered to all mice for 7 days. Samples of different tissues were collected on days 0, 3, and 7 post administration (DPA). High-iron treatment significantly decreased the relative weight of the large intestine at 7 DPA but not at 0 DPA or 3 DPA. High dietary iron increased the jejunal villus width at 0 DPA, decreased the villus width and the crypt depth of the jejunum at 3 DPA, and decreased the number of colonic crypts at 7 DPA. Meanwhile, high dietary iron decreased the number of goblet cells in the jejunal villi and the Paneth cells in the jejunal crypts at 0 DPA, increased the number of goblet cells per crypt of the colon at 3 DPA, and the number of Paneth cells in the jejunal crypts, the goblet cells in the colon, the Ki67-positive proliferating cells in the colon, and the Sex-determining region Y-box transcription factor 9+ (SOX9) cells in the jejunum crypts and colon at 7 DPA. The organoid formation rate was increased by high-iron treatments at 3 DPA and 7 DPA. High dietary iron treatment decreased the mRNA level of jejunal jagged canonical Notch ligand 2 (Jag-2) at 0 DPA and bone morphogenetic protein 4 (Bmp4) and neural precursor cell-expressed developmentally downregulated 8 (Nedd8) in the jejunum and colon at 7 DPA, whereas it increased the mRNA expression of the serum/glucocorticoid-regulated kinase 1 (Sgk1) in the colon at 3 DPA. The results suggested that a high dose of iron aggravated intestinal injury but promoted intestinal repair by regulating intestinal epithelial cell renewal and intestinal stem cell activity in adult mice with colitis.

Illegal fishing with electrofishing devices in the Po river basin, Emilia Romagna, Italy

Electric fishing is an illegal hunting method, unfortunately widely used by poachers to paralyze fish and to catch many animals in a short time. In Italy, it is authorized only for scientific and conservative purposes. Between 2014 and 2018, the Ferrara section of the Experimental Zooprophylactic Institute of Lombardy and Emilia Romagna, Italy, received nine cases of potentially illegal electric fishing in Po river and its tributary rivers. Necropsies were performed following standard protocols and samples of different tissues were collected and examined using histochemical and immunohistochemical techniques. Gross lesions frequently observed were circulatory alteration phenomena (i.e. multi-organ hyperemia, hemorrhages and congestion, hemopericardium), also found histologically, in addition to multifocal degenerative and necrotic muscular processes that could be attributed to injuries from electric current, as already reported in literature. Immunohistochemical investigations confirmed degenerative and necrotic lesions with myoglobin depletion and a corresponding fibrinogen accumulation. Myoglobin globules were also detected in the renal parenchyma, as consequent of rhabdomyolysis. The results of this study allowed to correlate electric fishing to gross, histologic and immunohistochemical lesions, which together constitute a pathognomonic picture to be considered a reference standard in this type of illegal controversy.

Occurrence and tissue distribution of perfluoroalkyl substances (PFASs) in sharks and rays from the eastern Mediterranean Sea

Persistent organic pollutants (POPs), including Perfluoroalkyl substances (PFASs), enter into the marine ecosystem, raising questions on possible adverse effects caused to the health of marine organisms and especially of top predators. Thus, there is an urge to assess the occurrence and the tissue distribution of PFASs in apex predators. To this end, the current study examines concentrations and distribution of 15 PFASs among 85 samples of different tissues from 9 shark and ray species collected in Greece. The results showed a similar PFAS pattern among the different tissues, with long carbon chain PFASs being the most frequently detected compounds. PFTrDA was the most predominant compound in terms of concentration and frequency of detection, followed by PFUnDA and PFOS. PFTrDA concentrations ranged between < LOQ and 27.1 ng/g ww, while PFUnDA and PFOS levels ranged from <LOQ to 16.0 and < LOQ to 21.6 ng/g ww, respectively. Regarding their frequency of detection, PFTrDA and PFUnDA were detected in 98% and 91% of the samples, respectively, while PFOS was detected in 79%. ΣPFAS concentrations in each analysed tissue ranged from 0.3 to 85 ng/g ww, with the latter being detected in the liver of angular roughshark (Oxynotus centrina). On average, PFASs were found to be accumulated in tissues in the following order: gonads > heart > liver ≈ gills > muscle. Relative contribution (%) of individual compounds to ΣPFAS concentration varied among the different shark tissues, and also among the different shark species. No correlation between PFASs levels in tissues and sharks’ gender, length and geographical origin was observed.

Abstract 5672: Facile generation of single-cell transcriptome and immune repertoire freshly isolated from clinical tumor specimens

Abstract Immunotherapies including cell-based therapies generate deep and durable responses in patients with chemotherapy-refractory cancers. However, in solid tumors, particularly those with stereotypic driver mutations and resultant neoantigens, the clonality and cell states of tumor-infiltrating lymphocytes (TILs) remain poorly understood. Here, we use a droplet-based 5' single-cell RNA-sequencing (scRNA-seq) to simultaneously profile transcriptome and immune repertoire of the same cells, enabling the phenotypic characterization (including cytotoxic and helper T, B, T regulatory and exhausted T cells) of each clonotype. We performed scRNA-seq on unsorted and CD45+-FACS sorted cells from fresh clinical samples of multiple tissue types, including lung, liver and kidney, and obtained on average tens of thousands of cells per sample. We demonstrated that high-quality single-cell suspension can be rapidly and reliably generated from clinical samples. We devised different diversity metrics to appropriately classify the immune repertoire within samples, and cross samples of different tissues. We observed distinct cell type compositions and, more importantly, context-specific T-cell clonal expansion patterns, suggesting the activation of different molecular programs in these tumors. In addition, comparison of non-small cell lung cancer (NSCLC) tumors varying by histology type and patient smoking status showed differences among these phenotypes, suggesting a potential link between immune microenvironments and cancer etiology. In summary, we provide a proof of concept for rapid generation of large number of single-cell transcriptomes of TILs paired with their corresponding TCR cDNA sequence in fresh tumor samples across different tissue types. Further analysis with this methodology on larger clinical cohorts will provide robust correlative prognostic markers of clinical phenotypes of immunotherapy responses. Our analysis strategy of TCR sequences among large clinical cohorts across multiple tumor types will facilitate cell-based therapeutic efforts, including CAR T cell or autologous T cell therapies. Citation Format: Junjie Zhu, Ameen A. Salahudeen, Valeria Giangarra, Luz Montesclaros, Jerald Sapida, Osman Sharifi, Josephine Lee, Grace X. Zheng, Dhananjay Wagh, John Coller, Chiara Sabatti, Calvin J. Kuo. Facile generation of single-cell transcriptome and immune repertoire freshly isolated from clinical tumor specimens [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5672.

Simultaneous determination of polysaccharides and 21 nucleosides and amino acids in different tissues of Salvia miltiorrhiza from different areas by UV-visible spectrophotometry and UHPLC with triple quadrupole MS/MS.

Salvia miltiorrhiza, a traditional Chinese medicine, is a widely used herbal medicine to treat cardiovascular and cerebrovascular diseases. In this study, ultraviolet (UV)-visible spectrophotometry and ultra-high performance liquid chromatography with triple quadrupole tandem mass spectrometry analytical methods were used for rapid quantification of polysaccharides and 21 nucleosides and amino acids in S. miltiorrhiza to determine 17 samples of different tissues from different areas. Based on the total contents, hierarchical clustering analysis and principal components analysis were performed to classify these samples. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Chemical analysis revealed a higher content of total analytes in the sample of inflorescence from Nanjing (34.17mg/g), sample of root and rhizome from Shaanxi (34.13mg/g) and sample of stem and leaf from Nanjing (31.14mg/g), respectively, indicating that root and rhizome from Shaanxi and the aerial parts from Nanjing exhibited the highest quality due to their highest content. In addition, contents of nucleosides and amino acids in the aerial parts (14.67mg/g) were much higher than that in roots and rhizomes (9.17mg/g). This study suggested that UV-visible spectrophotometry and ultra-high performance liquid chromatography with triple quadrupole tandem mass spectrometry are effective techniques to analyze polysaccharides, nucleosides, and amino acids in plants, and they provided valuable information for the development and utilization value of the aerial parts of S. miltiorrhiza. This analysis would also provide useful information for the quality control of S. miltiorrhiza.

Tissue distribution of ochratoxin A in pigs after administration of two-levels contaminated diets

The aim of the present study was to determine the levels of ochratoxin A (OTA) in pigs experimentally exposed to this mycotoxin and to evaluate if bile may be used to assess exposure to OTA. Twelve hybrid pigs were divided into 3 equal groups, a control group D0, and 2 experimental groups, D1 fed with 50 µg OTA/kg diet, and D2 fed with 500 µg OTA/kg diet for 15 days. At the end of the test, the animals were euthanized and samples of different tissues and biological fluids were analysed by HPLC-fluorescence detection for the presence of OTA. Samples of unconventional edible tissues such as lung and heart were also taken for analysis because they are used in typical Italian regional dishes. The Italian guidance value for OTA of 1 µg/kg established for pork meat and derived products has been exceeded in all the matrices from both the experimental groups. The comparison between OTA levels detected in D1 and D2 groups showed clearly a linear dose-response relationship. Based on the mean values measured, OTA distribution follows the order blood plasma &gt; lung &gt; kidney (in D1 group), heart (in D2 group) &gt; heart (in D1 group), kidney (in D2 group) &gt; bile &gt; liver &gt; fat &gt; muscle. Analysis of bile can be useful for the detection of OTA in pigs. However, since blood can easily be taken from pigs, and given the correlation between the mycotoxin concentration detected in this matrix and the concentrations detected in the others, OTA level in blood is a more viable approach to assessing the presence of OTA in edible tissues. As lung and heart may contain high concentrations of OTA, the analytical controls should also include these matrices.

Effects of thermal stress on the expression of glucocorticoid receptor complex linked genes in Senegalese sole (Solea senegalensis): Acute and adaptive stress responses

The present study examined the short and mid-term effects of a rise in temperature from 18°C to 24°C on the expression of genes related to the stress response regulation in juveniles of Senegalese sole, Solea senegalensis. The animals were exposed to a temperature increase of 6°C, after 1month of acclimation at 18°C. After this process, samples of different tissues were collected from a total of 96 fish at four sampling points: 1h, 24h, 3days and 1week. The transcript levels of a set of genes involved in the stress response such as glucocorticoid receptors 1 and 2, corticotrophin-releasing factor, corticotrophin-releasing factor binding proteins, proopiomelanocortin A and B, and cellular stress defense (heat shock protein 70, 90AA and 90AB) were quantified at these sampling points. Additionally, blood samples were also taken to measure the circulating plasma cortisol concentration.Thermal stress induced by increasing temperature prompted an elevation of plasma cortisol levels in juvenile Senegalese sole after 1h as a short-term response, and a consecutive increase after one week, as a mid-term response. Senegalese sole seemed to respond positively in terms of adaptive mechanisms, with a rapid over-expression of grs and hsps in liver and brain, significantly higher after one hour post stress, denoting the fast and acute response of those tissues to a rapid change on temperature. The ratio hsp90/gr also increased 24h after thermal shock, ratio proposed to be an adaptive mechanism to prevent proteosomal degradation of GR. As a mid-term response, the elevation of brain crfbp gene expression one week after thermal shock could be an adaptive mechanism of negative feedback on HPI axis.Taken together, these data suggested an initial up-regulation of the glucocorticoid receptor complex linked genes in response to a temperature increase in Senegalese sole, with heat shock protein 90 potentially being a regulatory factor for the glucocorticoid receptor in the presence of cortisol.

Outcome of oral infection in mice inoculated with Trypanosoma cruzi IV of the Western Brazilian Amazon

A new epidemiological view of American trypanosomiasis or Chagas disease has been formulated in recent decades. Oral transmission of the etiological agent of Chagas disease, Trypanosoma cruzi, has been the most common form of transmission. The T. cruzi discrete typing units TcI and TcIV have been involved in tens outbreaks of acute cases of Chagas disease in the Brazilian Amazon region. We investigated the intensity of infection in mice that were orally inoculated (OR group) with four strains of TcIV that were isolated from two outbreaks of acute Chagas disease that was orally acquired in the state of Amazonas, Brazil. We compared the OR group with mice that were intraperitoneally inoculated (IP group). Blood samples were analyzed by fresh blood examination, hemoculture, and conventional and qualitative real-time polymerase chain reaction (PCR). Samples of different tissues were analyzed by quantitative real-time PCR. The OR group exhibited a higher maximum peak of parasitemia, greater rates of positivity, and higher parasite loads in different tissues during acute infection compared with the IP group, indicating a greater intensity of orally acquired infection. Mice that were orally inoculated with TcIV strains that were obtained from two outbreaks of orally acquired Chagas disease in Amazonas, Brazil, exhibited a more intense course of infection compared with intraperitoneally inoculated mice, reflected by higher levels of parasitemia and parasite loads.

Post-mortem cytogenomic investigations in patients with congenital malformations

Congenital anomalies are the second highest cause of infant deaths, and, in most cases, diagnosis is a challenge. In this study, we characterize patterns of DNA copy number aberrations in different samples of post-mortem tissues from patients with congenital malformations. Twenty-eight patients undergoing autopsy were cytogenomically evaluated using several methods, specifically, Multiplex Ligation-dependent Probe Amplification (MLPA), microsatellite marker analysis with a MiniFiler kit, FISH, a cytogenomic array technique and bidirectional Sanger sequencing, which were performed on samples of different tissues (brain, heart, liver, skin and diaphragm) preserved in RNAlater, in formaldehyde or by paraffin-embedding. The results identified 13 patients with pathogenic copy number variations (CNVs). Of these, eight presented aneuploidies involving chromosomes 13, 18, 21, X and Y (two presented inter- and intra-tissue mosaicism). In addition, other abnormalities were found, including duplication of the TYMS gene (18p11.32); deletion of the CHL1 gene (3p26.3); deletion of the HIC1 gene (17p13.3); and deletion of the TOM1L2 gene (17p11.2). One patient had a pathogenic missense mutation of g.8535C>G (c.746C>G) in exon 7 of the FGFR3 gene consistent with Thanatophoric Dysplasia type I. Cytogenomic techniques were reliable for the analysis of autopsy material and allowed the identification of inter- and intra-tissue mosaicism and a better understanding of the pathogenesis of congenital malformations.

Virulence gene content in Escherichia coli isolates from poultry flocks with clinical signs of colibacillosis in Brazil

Escherichia coli is a commensal bacterium of the bird's intestinal tract, but it can invade different tissues resulting in systemic symptoms (colibacillosis). This disease occurs only when the E. coli infecting strain presents virulence factors (encoded by specific genes) that enable the adhesion and proliferation in the host organism. Thus, it is important to differentiate pathogenic (APEC, avian pathogenic E. coli) and non-pathogenic or fecal (AFEC, avian fecal E. coli) isolates. Previous studies analyzed the occurrence of virulence factors in E. coli strains isolated from birds with colibacillosis, demonstrating a high frequency of the bacterial genes cvaC, iroN, iss, iutA, sitA, tsh, fyuA, irp-2, ompT and hlyF in pathogenic strains. The aim of the present study was to evaluate the occurrence and frequency of these virulence genes in E. coli isolated from poultry flocks in Brazil. A total of 138 isolates of E. coli was obtained from samples of different tissues and/or organs (spleen, liver, kidney, trachea, lungs, skin, ovary, oviduct, intestine, cloaca) and environmental swabs collected from chicken and turkey flocks suspected to have colibacillosis in farms from the main Brazilian producing regions. Total DNA was extracted and the 10 virulence genes were detected by traditional and/or real-time PCR. At least 11 samples of each gene were sequenced and compared to reference strains. All 10 virulence factors were detected in Brazilian E. coli isolates, with frequencies ranging from 39.9% (irp-2) to 68.8% (hlyF and sitA). Moreover, a high nucleotide similarity (over 99%) was observed between gene sequences of Brazilian isolates and reference strains. Seventy-nine isolates were defined as pathogenic (APEC) and 59 as fecal (AFEC) based on previously described criteria. In conclusion, the main virulence genes of the reference E. coli strains are also present in isolates associated with colibacillosis in Brazil. The analysis of this set of virulence factors can be used to differentiate between APEC and AFEC isolates in Brazil.

Biomarkers: background, classification and guidelines for applications in nutritional epidemiology.

One of the main problems in nutritional epidemiology is to assess food intake as well as nutrient/food component intake to a high level of validity and reliability. To help in this process, the need to have good biomarkers that more objectively allow us to evaluate the diet consumed in a more standardized, valid and precise way has often been commented upon. There are various definitions of biomarkers and also different classifications of the same. In general a biomarker can be defined as a characteristic that can objectively measure different biological samples and that can be evaluated as an exposure marker of normal or pathogenic biological processes or of responses to a certain intervention. The biological samples most commonly used in nutritional epidemiology are blood, red blood cells, plasma, serum, urine, nails, saliva, faeces and samples of different tissues. Exposure biomarkers (dietary intake), biomarkers of effects and biomarkers of disease status can be determined from these samples. In turn, exposure biomarkers can be temporarily categorized into markers of acute, medium term or chronic effects. Many difficulties arise in identifying good biomarkers. Currently, advances in omics are opening up new possibilities for obtaining new biomarkers of various kinds, using genomics, epigenomics, transcriptomics, lipidomics, proteomics and metabolomics. We shall review the present situation of biomarkers in nutritional epidemiology as well as the future trends of the new omic biomarkers.

An investigation of the cause of death in Wistar rats exposed to sunlight: focus on tissue histology

Objective: The aim of the study is to investigate the effects of ultraviolet rays of the sun on organs like the liver, brain, heart, kidney, lung and ileum. Methods: Fourteen female rats (235 g) were used for the study and equally distributed into 2 groups. While the rats in the test groups were left in an open place for a period of 7 hours daily for adequate exposure to sunlight, control rats were kept in cages at ambient temperature of 26°C in the animal house. When all the animals died on the 10th day of study, samples of different tissues were obtained, fixed in 10% neutral buffered formalin and stained with hematoxylin and eosin (H & E). Results: 100% mortality was recorded. Results showed no visible lesion for all the tissues of control rats whereas pathological features like severe congestion of the coronary vessel and mild pulmonary congestion as well as thickening of alveolar wall were reported for heart and lung respectively in sun-exposed rats. Moreover, in sun-exposed rats other tissues such as the brain, ileum, kidney and liver showed no visible lesion. Conclusion: From the outcome of this study, it may be inferred that excessive sun exposure can be a cause of mortality in a nocturnal animal species, with death resulting from multi-organ damage.

Immunohistochemistry expression of TCF4 protein on carcinoma, adenoma and non neoplastic colorectal mucosa

PurposeTo detect and quantify the immunoreactivity of TCF4 protein in colorectal carci- noma, colorectal adenoma and non-neoplasic colorectal epithelium. MethodsWe studied 129 individuals: 40 with colorectal cancer, 52 with colorectal ad- enoma and 37 with non-neoplastic colorectal epithelium. The colorectal adenoma and carcinoma samples were obtained from patients who underwent surgical procedures, and colonoscopies and samples of non-neoplastic colorectal epithelium were taken from patients who died from cardiovascular diseases, without diseases of the large intestine. Samples of different tissues were included in paraffin blocks, and the immunohistochem- ical expression of protein TCF4 was analyzed using the technique of tissue microarray (TMA) with polyclonal antibody TCF4. The immunoreactivity was analyzed and classified as positive and negative. ResultsThe immunohistochemical expression of TCF4 protein was significantly higher (p < 0.01) in colorectal carcinoma than in the non-neoplastic colorectal epithelium and adenoma. There was no difference (p = 0.76) between TCF4 protein immunohistochemical expression in colorectal adenoma and non-neoplastic colorectal tissue. ConclusionsTCF4 protein showed a more intense expression in colorectal carcinoma than in non-neoplastic colorectal epithelium and adenoma, indicating that this protein is in- volved in colorectal carcinogenesis.

Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish Danio rerio and Jenynsia multidentata: In vitro studies

Although the herbicide glyphosate [N-(phosphonomethyl)glycine] is not classified as an acethylcholinesterase inhibitor, some studies have reported reduction in the acethylcolinesterase activity after in vivo exposure to both its pure form and its commercial formulations. Considering this controversy, the objective of the present study was to investigate, in vitro, the effects of glyphosate exposure on cholinesterase activity of the brown mussel Perna perna and of two fish species: zebrafish Danio rerio and onesided livebearer Jenynsia multidentata. For this purpose, samples of different tissues (brain and muscle for fish; gills and muscle for mussel) were homogenized and pre-incubated with different glyphosate concentrations before cholinesterase activity determination. Results demonstrated that cholinesterase from different fractions of all species tested was inhibited by glyphosate. The concentrations of glyphosate that inhibits 50% of cholinesterase activity (IC50) ranged from 0.62mM for P. perna muscle to 8.43mM for J. multidentata brain. According to this, cholinesterase from mussel seems to be more sensitive to glyphosate exposure than those from the fish D. rerio and J. multidentata.

Thermal Properties of Porcine Tissues Determined by Modified Photoacoustic Piezoelectric Technique

Using the modified photoacoustic piezoelectric (PAPE) technique, the influence of the piezoelectric transducer on the vibrations of the sample is taken into account. The modified PAPE technique is employed to determine the average thermal diffusivities of the porcine tissues, which include fresh and dry skin, fat, and muscle. The values of the thermal diffusivities of all measured porcine tissues determined by the modified PAPE technique are smaller than those of the conventional ones, especially for the dry skin and fresh fat samples. The thermal diffusivity of the fresh skin sample is the biggest, and the dry samples of different tissues have similar thermal properties with each other. These results show that the modified PAPE technique can provide thermal characterization of the porcine tissues more effectively.

Gene discovery using next-generation pyrosequencing to develop ESTs for Phalaenopsis orchids.

BackgroundOrchids are one of the most diversified angiosperms, but few genomic resources are available for these non-model plants. In addition to the ecological significance, Phalaenopsis has been considered as an economically important floriculture industry worldwide. We aimed to use massively parallel 454 pyrosequencing for a global characterization of the Phalaenopsis transcriptome.ResultsTo maximize sequence diversity, we pooled RNA from 10 samples of different tissues, various developmental stages, and biotic- or abiotic-stressed plants. We obtained 206,960 expressed sequence tags (ESTs) with an average read length of 228 bp. These reads were assembled into 8,233 contigs and 34,630 singletons. The unigenes were searched against the NCBI non-redundant (NR) protein database. Based on sequence similarity with known proteins, these analyses identified 22,234 different genes (E-value cutoff, e-7). Assembled sequences were annotated with Gene Ontology, Gene Family and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these annotations, over 780 unigenes encoding putative transcription factors were identified.ConclusionPyrosequencing was effective in identifying a large set of unigenes from Phalaenopsis. The informative EST dataset we developed constitutes a much-needed resource for discovery of genes involved in various biological processes in Phalaenopsis and other orchid species. These transcribed sequences will narrow the gap between study of model organisms with many genomic resources and species that are important for ecological and evolutionary studies.