falciparum IDC including H4K5ac, H4K8ac, H4K12ac, H4K16ac, H3K9ac

falciparum IDC including H4K5ac, H4K8ac, H4K12ac, H4K16ac, H3K9ac, H3K14ac, H3K56ac, H4K20me1, H4K20me3, H3K4me3, H3K79me3 and H4R3me2. While some modifications were found to be associated with the vast majority of the genome and their occupancy was constant, others showed more specific and highly dynamic distribution. Importantly, eight modifications displaying tight correlations with transcript levels showed differential affinity

to distinct genomic regions with H4K8ac occupying predominantly promoter regions while others occurred at the 59 ends of coding sequences. The promoter occupancy of H4K8ac remained unchanged when ectopically Selumetinib in vivo inserted at a different locus, indicating the presence of specific DNA elements that

recruit histone modifying enzymes regardless of their broad chromatin environment. In addition, we showed the presence of multivalent domains on the genome carrying more than one histone mark, highlighting the importance of combinatorial effects Compound C on transcription. Overall, our work portrays a substantial association between chromosomal locations of various epigenetic markers, transcriptional activity and global stage-specific transitions in the epigenome.”
“PURPOSE: To evaluate the effectiveness of radiologic placement of uncovered stents for the treatment of malignant colorectal obstruction.\n\nMATERIALS AND METHODS: From May 2003 to January 2008, 116 radiologic placements of uncovered stents were attempted in 99 patients (M:F, 59:40; mean age, 65 years) with malignant colorectal obstructions. The location of stent insertion, technical and clinical success, complication

rates, and patency rates of the stents in a palliative AZD8055 clinical trial group were also evaluated. In the palliative group, the follow-up period was 2-455 days (mean, 100 +/- 129 days).\n\nRESULTS: Radiologic stent placement was technically successful in 110 of 116 cases (94.8%). Fifty cases of stent placement were preoperative (45.5%, 50 of 110) and 60 (54.5%, 60 of 110) were performed with palliative intents. In five of six failed cases, the replacement of the stent was later performed with the assistance of colonoscopy. One patient underwent an emergency operation. In 98 of 110 cases, the symptoms of obstruction were relieved, for a clinical success rate of 89.1%. Of the 50 stents that were placed successfully with preoperative intent, 44 patients underwent surgery within a mean of 10.3 days. In the palliative group, the patency rates were 89.7% at 1 month, 85.6% at 3 months, 80.8% at 6 months, and 72.7% at 12 months.\n\nCONCLUSIONS: The radiologic placement of uncovered stents for the treatment of malignant colorectal obstruction is feasible and safe and provides acceptable clinical results not only for preoperative decompression but also for palliative cases, especially in left-sided colonic obstructions.

YCE were

YCE were PP2 ic50 obtained at different pH (4, 7 and 10) and ultra-filtered 300 and 30 kDa membranes (UYE). The 30 kDa UYE was heated to 60 degrees C, 90 degrees C and 120 degrees C and gel filtered (GYF). Mid infrared spectroscopy, protein and carbohydrate analysis of GYF were conducted. Results showed that YCE, UYE and GYF significantly stimulated (p < 0.05) the biomass production, acetate concentration and carboxymethyl cellulase activity of the ACC, in relation to the control. The GYF had an estimated molecular mass of 4 kDa. Mid-infrared and biochemical analysis of GYF suggested

that the active compound is a peptide.”
“Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common

forms of inheritable Parkinson’s disease and likely play a role in sporadic disease as well. LRRK2 is a large multidomain protein containing two key groups, a Ras-like GTP binding domain and a serine, threonine kinase domain. Mutations in the LRRK2 gene that associate with Parkinson’s disease reside primarily within the two functional domains of the protein, suggesting that LRRK2 function is critical to the pathogenesis of the disease. The most common LRRK2 mutation increases kinase activity, making LRRK2 kinase inhibition an attractive target for small molecule drug development. However, the physiological function of LRRK2 kinase as well as its endogenous protein substrates remains poorly understood and has hindered drug development efforts. Recent advances in LRRK2 biology have revealed several potential cellular roles, GSK3326595 purchase interacting Adavosertib mouse proteins, and putative physiological substrates. Together, a picture emerges of a complex multifunctional protein that exists in multiple cellular compartments. Through unclear mechanisms, LRRK2 kinase regulates cytoskeleton architecture through control of protein

translation, phosphorylation of cytoskeletal proteins, and response to cellular stressors. This article will briefly cover some interesting recent studies in LRRK2 cellular biology and highlight emerging cellular models of LRRK2 kinase function.</.”
“Introduction: This article discusses findings from an ethnographic study of a national policy initiative in Wales, United Kingdom. The policy aimed to decrease the caesarean section rate and increase the number of normal births by implementing a clinical pathway for normal labour. The aim of this study was to explore the real life experiences of those involved in the development and use of the policy.\n\nMethods: A range of qualitative methods was used, including observation, semistructured interviews, focus groups, and documentary analysis. Data were collected from midwives, doctors, midwifery managers, and mothers. Field notes and transcripts were thematically analysed.

In addition, the results suggest that specific cognitive domains,

In addition, the results suggest that specific cognitive domains, particularly

executive functions, are likely to have the strongest effect on functional outcomes.”
“Objective. High-risk human papillomavirus (HR-HPV) persistence is thought to be necessary for the development of cervical cancer. Because not all cases of low-grade squamous intraepithelial lesion (LSIL) or higher-grade squamous intraepithelial lesion are associated with HR-HPV, detection of HR-HPV in women with these cervical abnormalities may be clinically useful for management. The aim of this study was to detect HR-HPV in women with LSIL or higher-grade cytology.\n\nMaterials and Methods. In a multicenter, prospective clinical study, residual liquid-based Cl-amidine mouse cytological specimens from 3,966 US women were analyzed. Women with LSIL or higher-grade cytology underwent colposcopy and cervical biopsy as clinically indicated. The test was used to detect HR-HPV in women with cytologically diagnosed LSIL or higher, with histological confirmation of cervical intra-epithelial neoplasia www.selleckchem.com/products/Gefitinib.html 2 (CIN 2) or worse by a central review panel.\n\nResults. Among subjects with LSIL or higher-grade cytology and complete data sets, 11.8% (41/347) were diagnosed with CIN 2 or worse; 82.1% (285/347) were

HR-HPV positive. The prevalence of CIN 2 or worse and CIN 3 or worse was 14.4% (41/285) and 7.0% (20/285), respectively, among subjects who were HR-HPV positive. All subjects diagnosed with CIN 2 or worse (41/41) and CIN 3 or worse (20/20) tested positive for HR-HPV. Sensitivity and negative predictive value calculated for CIN 2 or worse were 100% (95% Cl = 91.4%-100.0%) and 100% (95% Cl = 94.2%-100.0%), respectively. Specificity and positive predictive value among these subjects were 20.3% (95% Cl = 16.1%-25.1%) and 14.4% (95% Cl = 10.8%-18.9%), respectively.\n\nConclusions.

Detection of HR-HPV in women with LSIL or higher-grade cytology using the HPV HR test may allow clinicians to further triage women for advanced clinical management.”
“Background: Fluralaner is a novel systemic insecticide and acaricide that provides long acting efficacy in dogs after Crenigacestat chemical structure a single oral treatment. This study investigated the safety of oral administration of fluralaner in chewable tablets to dogs at the highest recommended treatment dose and at multiples of this dose. Methods: Thirty-two (16 male and 16 female) healthy 8-week old Beagle dogs weighing 2.0 -3.6 kg at first administration were included in the study. Fluralaner was administered on three occasions at 8-week intervals at doses of up to 56, 168, and 280 mg fluralaner/kg body weight, equivalent to 1, 3, and 5 times the highest recommended treatment dose of fluralaner; sham dosed dogs served as controls.

(C) 2011 Elsevier Ltd All rights reserved “
“A systematic l

(C) 2011 Elsevier Ltd. All rights reserved.”
“A systematic literature review was performed regarding the burden on mental health caregivers. The studies were selected from the Virtual Health Library – Biblioteca Virtual de Sa de (BVS), using the keyword caregiver burden. The main criteria for this study were: full-text articles published between 2000 and 2010, in Portuguese, English or

Spanish; indexed on the BVS databases; which investigated the burden of mental health caregivers, and had caregivers as the main subject. The analysis was performed considering the following: title, year of publication, objectives, methodological approach, instruments and main results. The analysis of 114 full-text articles showed the predominant p53 inhibitor objectives were the burden on

informal caregivers and the validation see more of psychometric scales, particularly the Zarit Scale. Some studies showed an association between high levels of burden, feelings of guilt and depressive symptoms. On the other hand, psycho-educational interventions were indicated as having a positive impact. This theme has a growing scientific interest and there is a need for deeper studies addressing formal caregivers.”
“PurposeTo study balanced steady-state free precession CINE phase-sensitive water-fat separation imaging in four cardiac imaging planes to determine the necessary phase correction and image artifacts particular to this technique. MethodsTen healthy volunteers and two subjects with known heart pathologies were studied with standard balanced steady-state free precession CINE imaging.

Water-only and fat-only images were calculated using sign detection of the real part of the complex image after phase correction with constant and linear terms. Phase correction values were determined using both manual and automated methods. Differences in phase correction values between imaging planes, cardiac phases, coil elements, automated image reconstruction parameters as well as artifact scores between the automated and manual methods were studied with statistical tests. ResultsWater-fat separation performed well in the heart after constant and linear phase correction. Both constant (p = 0.8) and linear x (p = 1) and y (p = 1) phase correction values did not vary significantly across cardiac phases, but CP-868596 cell line varied significantly among the coils (p smaller than 0.001) and imaging planes (p smaller than 0.001). False water-fat separation artifacts were most frequent in the chest/back and also were present at the mitral and aortic valves. ConclusionConstant and linear phase correction is necessary to provide consistent results in standard imaging planes using a balanced steady-state free precession water-fat separation postprocessing algorithm applied to standard cardiac CINE imaging. Magn Reson Med 71:2096-2104, 2014. (c) 2013 Wiley Periodicals, Inc.

HAT is a challenging and deadly disease owing to its complex epid

HAT is a challenging and deadly disease owing to its complex epidemiology and clinical presentation Cell Cycle inhibitor and, if left untreated, can result in high death rates. As one of the most neglected tropical diseases, HAT is characterized by the limited availability of safe and cost-effective control tools. No vaccine against HAT is available, and the toxicity of existing old and cumbersome drugs precludes the adoption of control strategies

based on preventive chemotherapy. As a result, the keystones of interventions against sleeping sickness are active and passive case-finding for early detection of cases followed by treatment, vector control and animal reservoir management. New methods to diagnose and treat patients and to control transmission by the tsetse fly are needed to achieve the goal of global elimination of the disease.”
“To make an antisaccade away from a stimulus, one must also suppress the more reflexive prosaccade to the stimulus. Whether this inhibition is diffuse or specific for saccade direction is not known. We used a paradigm examining inter-trial carry-over effects. Twelve subjects performed sequences of four identical antisaccades followed by sequences of four prosaccades randomly directed at the location of the antisaccade stimulus, the location of the antisaccade goal, or neutral locations.

We found two types of persistent antisaccade-related inhibition. Epigenetics inhibitor First, prosaccades in any direction were delayed only in the first trial after the antisaccades. Second, prosaccades to the location of the antisaccade stimulus were delayed more than all other prosaccades, and this persisted from the first to the fourth subsequent trial. These Rigosertib solubility dmso findings are consistent with both a transient global inhibition and a more sustained focal inhibition

specific for the location of the antisaccade stimulus. (C) 2011 Elsevier B.V. All rights reserved.”
“Clostridium difficile is mainly associated with nosocomial infections but can be present also in other environments. In this study we compared three methods (culturing with and without ethanol shock and real-time PCR) for detection of C. difficile in water and have used them on a series of river water samples. C. difficile was present in 17 of 25 rivers tested (68.0%) and in 42 of 69 water samples tested (60.9%). Positive sampling sites correlated with increased population densities. Isolates were distributed into 34 PCR ribotypes, of which more than half are present also in humans and animals. PCR ribotype 014 was the predominate type (16.2% of all isolates). (C) 2010 Elsevier Ltd. All rights reserved.”
“Sediments from Xinyun Lake in central Yunnan, southwest China, provide a record of environmental history since the Holocene.

The models account for the geometry of MPs and heterogeneous dist

The models account for the geometry of MPs and heterogeneous distribution of membrane channels and receptors in an EC. center dot Simulations show that SMC stimulation causes calcium release in and around EC MPs that activates hyperpolarizing currents in ECs and moderates SMC constriction. center dot The results help us better understand the mechanisms that regulate selleck kinase inhibitor blood flow and pressure. Abstract We investigated the role of myoendothelial projections (MPs) in endothelial cell (EC) feedback response to smooth muscle cell (SMC) stimulation using mathematical modelling. A previously developed compartmental

EC-SMC model is modified to include MPs as subcellular compartments in the EC. The model is further extended into a 2D continuum model using a finite element method (FEM) approach and electron microscopy images to account for MP geometry. The EC and SMC are coupled via non-selective myoendothelial gap junctions (MEGJs) which are located on MPs and allow exchange of Ca2+, K+, Na+ and Cl- ions and inositol 1,4,5-triphosphate (IP3). Models take into consideration recent evidence for co-localization of intermediate-conductance calcium-activated potassium channels (IKCa) and IP3 receptors (IP3Rs) in the MPs. SMC stimulation

causes an IP3-mediated Ca2+ transient in the MPs with limited global spread in the bulk EC. A hyperpolarizing feedback generated by the localized IKCa channels is transmitted to the SMC via MEGJs. MEGJ resistance (Rgj) and the density of IKCa and IP3R in the projection influence the extent of EC response to SMC stimulation. Momelotinib cell line The predicted Ca2+ transients depend also on the volume and geometry of the MP. We conclude that in the myoendothelial feedback response to SMC stimulation, GSK1120212 research buy MPs are required to amplify the SMC initiated signal. Simulations suggest that the signal is mediated by IP3 rather

than Ca2+ diffusion and that a localized rather than a global EC Ca2+ mobilization is more likely following SMC stimulation.”
“Vascular tumor is an abnormal buildup of blood vessels in the skin or internal organs that can lead to disfigurement and/or life-threatening consequences. The mechanism of hemangiogenesis remains unknown. The aim of this study was to assess the role of rapamycin-insensitive companion of mTOR (Rictor) in control of vascular tumor malignant biological behavior and cell signaling mechanism in Mouse Hemangioendothelioma Endothelial Cells (EOMA cells) and nude mouse model. Knocking down rictor was mediated by lentivirus shRNA. The role and mechanism of rictor in vascular tumor were assessed by western blotting, wst-1 proliferation assay, matrigel invasion assay and xenograft vascular tumor growth. Our results in vitro showed that loss of rictor down-regulated phosphorylation of AKT and S6 by which EOMA cells growth and proliferation were greatly suppressed. Knock down of rictor also inhibited the invasion of EOMA cells.

We previously found that the I kappa B kinase

beta (IKK b

We previously found that the I kappa B kinase

beta (IKK beta)/NF-kappa B pathway regulates early IFN-beta expression, but not the magnitude of type 1 IFN expression following RLR engagement. In this study, we use IKK beta inhibition and mice deficient in IKK beta or canonical NF-kappa B subunits (p50, RelA/p65, and cRel) to demonstrate that the IKK beta/NF-kappa B axis is critical for virus-induced type 1 IFN expression in pDCs, but not in cDCs. We also reveal a crucial and more general requirement for IKK beta/NF-kappa B in TLR- but not RLR-induced expression of type 1 IFNs and inflammatory cytokines. Together, these findings reveal a previously unappreciated specificity of the IKK beta/NF-kappa Ruboxistaurin chemical structure B signaling axis in regulation of antimicrobial responses by different classes of pattern recognition receptors, and therefore by individual P005091 solubility dmso cell types reliant on particular pattern recognition receptors for their innate-immune transcriptional responses.”
“Recent studies have reported that subclinical hypothyroidism (SCH) is associated with atherosclerosis (AS). Thyroid hormone is maintained at normal levels in patients

with SCH, whereas TSH is increased. However, the pathogenesis of AS in association with SCH is only partially understood. In addition, endothelial dysfunction plays an important role in the development of AS. The purpose of the present research was to study the direct effect of TSH on human umbilical vein endothelial cells (HUVECs). The expression of some genes associated with endothelial dysfunction after treatment with TSH was evaluated by real-time PCR and western blotting respectively. At first, we showed that the TSH receptor (TSHR) is expressed in HUVECs. We also provide evidence indicating that TSH treatment promotes tumor necrosis factor alpha-induced endothelial cells interactions by upregulating the expression of the adhesion

molecules intercellular adhesion molecule-1. Furthermore, the expression of endothelial nitric oxide synthase (eNOS) and prostacyclin (PGI(2)) was significantly attenuated this website following treatment with TSH in dose- and time-dependent manner. Conversely, the results indicated that TSH upregulated endothelin-1 (ET1) mRNA and protein expression in HUVECs, similar effects were observed for plasminogen activator inhibitor-1 (PAI1) after treatment with various concentrations of TSH. Taken together, these results demonstrate that elevated TSH can promote endothelial dysfunction by altering gene expression in HUVECs.”
“Background: Specific centile growth charts for children with Down syndrome (DS) have been produced in many countries and are known to differ from those of normal children.

We tested for the effect of hydrological regime on the structure

We tested for the effect of hydrological regime on the structure and functions of microbial communities by comparing sediments collected at multiple sites in the Ain departement (Eastern France). Each site consisted of two plots, one permanently and one seasonally inundated. At the time of sampling, all plots were continuously inundated for more than 6months but still harboured distinct bacterial communities. This change in community structure was not associated with marked modifications in the rates

of microbial activities involved in the C and N cycles. These results suggest that the observed structural change could be related to bacterial taxa responding to the environmental variations associated with different hydrological regimes, but Adavosertib molecular weight not strongly associated with the biogeochemical processes monitored here.”
“BackgroundCutaneous squamous cell carcinoma

(cSCC) usually has ill-defined margins because of its irregular invasive patterns. ObjectiveTo evaluate the surgical efficacy of photodynamic diagnosis (PDD) in primary cSCC treated using Mohs micrographic surgery (MMS). Methods & MaterialsWe examined 67 cases of biopsy-proven primary facial cSCC treated with MMS. The 67 SCC were divided into the two groups depending on PDD application: PDD group (n=38, 56.7%) and non-PDD group (n=29, 43.3%). We analyzed the differences in surgical features between the PDD and non-PDD groups. ResultsThe PDD group required fewer Mohs stages (1.37 vs 1.83, LY3039478 datasheet p=.02) and smaller surgical margins (8.03 vs 11.24mm, p=.03). PDD showed additional benefits in terms of surgical margin and Mohs stage, especially in low-risk SCC, including thin (4mm), small buy Fedratinib (20mm), well-differentiated, and nonulcerative tumors (p smaller than .05) but did not show beneficial effects in high-risk SCC (p bigger than .05) during MMS. ConclusionPDD

can increase surgical efficacy of primary cSCC during MMS. These benefits are more pronounced in low-risk SCC. We recommend PDD as a simple and useful technique for delineating the margins of low-risk SCC before MMS.”
“Impacting hardenable steel such as 4340, results in the formation of adiabatic shear bands (ASBs). Previous studies have shown that the presence of carbides/second-phase particles in the pre-deformation microstructures of 4340 steel increases their susceptibility to the formation of ASBs. The current study examines the role of carbides on the microstructure and properties within evolved ASBs in 4340 steel after impact. Geometric phase analysis was used to map local deformation fields within the evolved ASBs. It was observed that carbide fragmentation due to plastic deformation of carbides produces both residual carbides and residual carbide particles in regions away from the shear bands.

Characteristics of vision loss in this family include early chron

Characteristics of vision loss in this family include early chronic optic nerve edema, and progressive vision loss, particularly central and color vision. Despite numerous medical and ophthalmic evaluations, no diagnosis has been discovered. (C) 2012 Wiley Periodicals, Inc.”
“Background: We have previously reported that cancer incidence for

lung, female breast, and colon and rectum for Hispanics decreases with increasing percentage of Hispanics at the census tract. In contrast, cervical cancer incidence increases click here with increasing percentage of Hispanics at the census tract.\n\nMethods: In this study, we investigate the hypothesis that Hispanics living in census tracts with high percentages of Hispanics are diagnosed with more advanced cancer, with respect to tumor size and stage of diagnosis. Data from the Surveillance, Epidemiology, and End Results registry and the U.S. Census Bureau were used to estimate the odds of diagnosis at a “late” stage (II, III, IV) versus “early” stage (1) and breast cancer tumor size among Hispanics as a function of census tract percent Hispanic. Hispanic ethnicity in the Surveillance, Epidemiology, and End Results registry was identified by medical record review and Hispanic surname lists. The study also used income of Hispanics living

in the census tract and controlled for age at diagnosis and gender.\n\nResults: We found that Hispanics living in neighborhoods SBE-β-CD order with higher density of Hispanic populations were more likely see more to be diagnosed with late-stage breast, cervical, or colorectal cancer, and to have a larger

tumor size of breast cancer.\n\nConclusions: Our findings suggest that the benefits of lower cancer incidence in high tract percent Hispanics are partially offset by poorer access and reduced use of screening in conjunction with lower income, poorer health insurance coverage, and language barriers typical of these communities. (Cancer Epidemiol Biomarkers Prev 2008;17(11):2931-6)”
“Evolution of proteins involves sequence changes that are frequently localized at loop regions, revealing their important role in natural evolution. However, the development of strategies to understand and imitate such events constitutes a challenge to design novel enzymes in the laboratory. In this study, we show how to adapt loop swapping as semiautonomous units of functional groups in an enzyme with the (beta/alpha)(8)-barrel and how this functional adaptation can be measured in vivo. To mimic the natural mechanism providing loop variability in antibodies, we developed an overlap PCR strategy. This includes introduction of sequence diversity at two hinge residues, which connect the new loops with the rest of the protein scaffold, and we demonstrate that this is necessary for a successful exploration of functional sequence space.

After analyzing the evidence, we hypothesize the need to prime th

After analyzing the evidence, we hypothesize the need to prime the immune system to restore natural tolerance against a-synuclein in Parkinson disease, including at the same time B and T cells, so that T cells can reprogram microglia activation to a beneficial pattern and B cell/IgG can help neurons cope with the pathological forms of a-synuclein.”
“The role played

by apoptosis in the homeostasis of effector cells of the innate immune system is unclear. Serine protease inhibitor 6 (Spi6) is an inhibitor of granzyme B (GrB) that protects cytotoxic T lymphocytes of the adaptive CA4P molecular weight immune system from apoptosis. To determine whether Spi6 also protects cells of the innate immune system from self-inflicted damage we have examined invariant NKT (iNKT) cells. Spi6-deficient iNKT cells harbored increased levels of GrB after TCR stimulation with the PBS-57 glycolipid Ag and were susceptible to apoptosis. The increased apoptosis of Spi6 knock-out (KO) iNKT cells lead to a complete loss in the production of IL-4 and IFN-gamma by Spi6 KO iNKT cells after PBS-57 challenge. The increased activation-induced apoptosis resulted in impaired survival and a decreased clonal burst size of Spi6 KO iNKT cells, which could be corrected

by GrB deficiency. However, the clonal burst of Spi6 KO iNKT cells after TCR-independent activation with lymphocytic choriomeningitis virus was not affected. Our findings demonstrate that Spi6 protects cytotoxic cells of the innate immune system from GrB-mediated self-inflicted triggered by the recognition of Ag. GSK J4 The Journal of Immunology, 2010, 185: 877-883.”
“Suppressor of cytokine signalling (SOCS) proteins are inhibitors of cytokine signalling pathways. Three SOCS genes, SOCS-1, 2 and 3, have been identified and their sequences analyzed in an economically important fish, rainbow trout (Oncorhynchus mykiss, Walbaum). In general, these

three SOCS molecules are well conserved especially in the SRC homology 2 and the SOCS domains, with sequence identities between Ganetespib order trout and mammals ranging from 41 to 42, 50 to 51, and 58 to 61% for SOCS-1, 2 and 3, respectively. The identities within fish species are slightly higher, with sequence identities between trout and the other fish species at 44-46, 64-70, and 71-76% for SOCS-1, 2 and 3, respectively. All the SOCS-1, as well as all the SOCS-2 or 3 molecules from different species are grouped together in phylogenetic tree analysis with high bootstrap support, with the fish molecules in each type grouping closely together. The expression of the trout SOCS-1, 2 and 3 genes are detectable by real-time PCR in all the eight tissues studied; the gills, skin, muscle, liver, spleen, head kidney, intestine and brain. SOCS-1 is highly expressed in intestine, head kidney, spleen, gills and skin. SOCS-2 is highly expressed in brain, head kidney, muscle, spleen, gills, skin and intestine.