Tissue Sections Of Organs Research Articles

Protocol refinement for quenching autofluorescence of red blood cells in FFPE sections of organ samples from cattle, pigs and chickens

One of the most common problem that researchers encounter when using fluorescence to visualize immunohistochemistry is the autofluorescence of the studied organ tissue sections and cell cultures. Autofluorescence quenching is necessary for a wide variety of organs and tissues, as well as for different methods of fixation and histochemical processing of sections. In addition to autofluorescence quenching, it is necessary to take into account the need for histological readability of tissue sections when using counterstains afterwards. Such protocol refinement for fluorescent immunohistochemistry for chicken, porcine and cattle tissues was carried out for the first time, as well as the use of a dimethyl sulfoxide (DMSO) solution with ethanol as Sudan Black B (SBB) solvent. Incubation of sections in SBB was chosen as the simplest and most nonspecific one. The most effective dissolution of the dye is achieved at a concentration of 0.3% SBB in a solution of 70% ethanol and absolutized DMSO in a 4:1 v/v ratio. The most thorough removal of SBB solution excess is achieved by rinsing the sections 5 times with 70% ethanol and then rinsing the sections with TBST (tris-buffered saline and Tween-20) buffer 5 times.

Study the Histopathological Effect of Aristolochia bracteolata L. on Common Freshwater Carp Oreochromis Niloticus

Abstract: Effects of water-soluble extracts of the fresh mature leaves of Aristolochia bracteolata were studied on common freshwater carp Oreochromis niloticus and observed the Histopathological effect on different tissues, including liver, kidney, and intestine in the laboratory. First, select the two fishes for the experiment. Then, each fish was placed in a glass aquarium containing 10 litres of dechlorinated tap water and named treated and controlled. The 1.5 ml leaf extract exposed fish in the first aquarium; after that, the to and fro movement increased. At the end of the experiment, the mortality of fish occurred. Next, take the tissue sections of organs from both the fishes and treat and stain with haematoxylin and eosin. The stained slides were observed under a compound microscope at 45X and recorded in different observations. Finally, we concluded that the exposure of aqueous leaf extract of Aristolochia bracteolata L. resulted in apparent toxic effects on the kidneys of common carp Oreochromis niloticus.

Evaluation of the Safety and Gastrointestinal Migration of Guanidinylated Chitosan after Oral Administration to Rats.

Arginine-rich membrane-permeable peptides (APPs) can be delivered to cells by forming complexes with various membrane-impermeable bioactive molecules such as proteins. We recently reported on the preparation of guanidinylated chitosan (GCS) that mimics arginine peptides, using chitosan, a naturally occurring cationic polysaccharide, and confirmed that it enhances protein permeability in an in vitro cell system. However, studies on the in vivo safety of GCS are not available. To address this, we evaluated the in vivo safety of GCS and its translocation into the gastrointestinal tract in rats after a single oral administration of an excessive dose (500 mg/kg) and observed changes in body weight, major organ weights, and organ tissue sections for periods of up to 2 weeks. The results indicated that GCS causes no deleterious effects. The results of an oral administration of rhodamine-labeled chitosan and an evaluation of its migration in the gastrointestinal tract suggested that the disappearance of rhodamine-labeled GCS from the body appeared to be slower than that of the non-dose group and pre-guanidinylated chitosan due to its mucoadhesive properties. In the future, we plan to investigate the use of GCS to improve absorption using Class III and IV drugs, which are poorly water-soluble as well as poorly membrane-permeable.

Effect of Calcitonin Hormone of Biochemical And Histological Changes for Female Sailfin Black Molly (Poecilia latipinna)

Introduction: Calcitonin (CT) regulates blood calcium (Ca+2) levels in mammals, but the results of experiments on its role in fish are uncertain, its functions exceeded the limits of regulation of Ca+2.Aim: This experience represents a trial to remove that ambiguity and determine role of calcitonin by clarifying its relationship with different biochemistry parameters. Therefore, we investigate the effect of (CT) on some biochemical parameters of blood in female fish Poecilia latipinna, these are Ca+2, urea, creatinine, alkaline phosphatase enzyme (ALP), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). The study also deals with the follow up of histological changes that are caused by (CT) in the liver, kidney, and intestines of the fish.Material and Methods: The fish were brought from ornamental fish shops and were adapted and reared in the laboratory. The sample of fish was divided into two groups; The first group was injected with 4 microliters of (CT) for ten consecutive days. At the end of the experiment, blood was drawn from the fish and then it anatomized and tissue sections of organs under study were prepared.Result: The finding show calcitonin works to reduce level of Ca+2 in the blood with a significant difference (0.03) at the probability level P ≤ 0.05. And was not affected urea and creatinine by the hormone, while the level of ALP enzyme increased with a significant difference (0.04) at the probability level P ≤ 0.05. With sex hormones, the effect was limited to LH without FSH. LH levels increased significantly with (0.02) at P ≤ 0.05. CT caused histological changes in the kidney and intestines, while the liver was not affected by (CT).Conclusion: Calcitonin has been shown to play a role in Ca+2 regulation in fish blood and its work overlaps with different organs of the fish body.

Novel contrast media based on the liquid metal gallium for in vivo digestive tract radiography: a feasibility study

The barium sulfate has been playing an important role as the contrast medium in gastrointestinal radiography and disease diagnosis. However, its application has been gradually reduced due to the limitation of its imaging effect and the progress of other imaging techniques. Here, the liquid metal gallium was proposed as an improved contrast agent, which was applied in the in vivo digestive tract radiography of the mice for the first time. Under the CT scanning, the gallium produced excellent contrast effect intuitively. According to the records of discharge time, the tissue sections of organs, the survival state and body weight, the liquid metal was proven to be capable and safe for gastrointestinal radiography. Further, the mixture of the gallium and the barium sulfate has been tested, which showed better performance in both contrast and detail. Therefore, with the characteristics of better imaging contrast effect and acceptable safety, the gallium and its mixture with the barium sulfate might be useful as potential candidates for digestive tract contrast agent in animal experiments, even possibly as alternative contrast agents for clinical use.

Evaluation of antiviral effect and toxicity of total flavonoids extracted from Robinia pseudoacacia cv. idaho

In this study, we aimed to evaluate the antiviral effect of total flavonoids extracted from Robinia pseudoacacia cv. idaho (RPTF) in vivo and its toxicity on rats with oral gavage. RPTF was prepared by percolation with 70% ethanol for 24 h and its antiviral effect on different kinds of viruses was evaluated in vitro by MTT staining. The long-term toxicity of RPTF on rats was evaluated through the detection of general behavior, body weight, food intake and related organ tissue sections of experimental animals. We found that RPTF produced significantly inhibitory effects on HSV-1 and EV-71 viruses with the therapeutic index TI values 113.8 and 46.2, respectively. Moreover, toxicity evaluation in vivo showed no significantly adverse effects in rats, indicating that RPTF was safe in use. In conclusion, we demonstrated that RPTF, natural compounds in the Chinese traditional medicine, could act as promising and effective antiviral therapeutics with relative safety in use.

Carbon nanotube translocation to distant organs after pulmonary exposure: insights from in situ (14)C-radiolabeling and tissue radioimaging.

Few approaches are available to investigate the potential of carbon nanotubes (CNTs) to translocate to distant organs following lung exposure, although this needs to be taken into account to evaluate potential CNT toxicity. Here, we report a method for quantitative analysis of the tissue biodistribution of multiwalled CNTs (MWCNTs) as a function of time. The method relies on the use of in situ (14)C-radiolabeled MWCNTs and combines radioimaging of organ tissue sections to ex vivo analysis of MWCNTs by electron microscopy. To illustrate the usefulness of this approach, mice were exposed to a single dose of 20 μg of (14)C-labeled MWCNTs by pharyngeal aspiration and were subjected to a follow-up study over one year. After administration, MWCNT were cleared from the lungs, but there was a concomitant relocation of these nanoparticles to distant organs starting throughout the follow-up period, with nanoparticle accumulation increasing with time. After one year, accumulation of MWCNTs was documented in several organs, including notably the white pulp of the spleen and the bone marrow. This study shows that the proposed method may be useful to complement other approaches to address unresolved toxicological issues associated with CNTs. These issues include their persistence over long periods in extrapulmonary organs, the relationship between the dose and the extent of translocation, and the effects of "safety by design" on those processes. The same approach could be used to study the translocation propensity of other nanoparticles containing carbon atoms.

Specificity of antibodies established from mammals in rainbow trout (Oncorhynchus mykiss)

Summary This study deals with the question as to whether antibodies established for mammals are also specific for rainbow trout. The reason for this examination is the major economic importance of rainbow trout in aquaculture and the growing scientific attention. However, there are few primary antibodies so far defined for this fish species. Therefore, the aim of the current study was to test the ability of 15 commercially available antibodies for rainbow trout in an indirect immunofluoresence assay to analyse tissue sections of organs. Five commercially available primary antibodies were identified, which were directed against proteins of one or two organs/ cell types in rainbow trout.

Detection of specific glycosaminoglycans and glycan epitopes by in vitro sulfation using recombinant sulfotransferases

Sulfated glycans play critical roles during the development, differentiation and growth of various organisms. The most well-studied sulfated molecules are sulfated glycosaminoglycans (GAGs). Recent incidents of heparin drug contamination convey the importance of having a convenient and sensitive method for detecting different GAGs. Here, we describe a molecular method to detect GAGs in biological and biomedical samples. Because the sulfation of GAGs is generally not saturated in vivo, it is possible to introduce the radioisotope (35)S in vitro using recombinant sulfotransferases, thereby allowing detection of minute quantities of these molecules. This strategy was also successfully applied in the detection of other glycans. As examples, we detected contaminant GAGs in commercial heparin, heparan sulfate and chondroitin samples. The identities of the contaminant GAGs were further confirmed by lyase digestion. Oversulfated chondroitin sulfate was detectable only following a simple desulfation step. Additionally, in vitro sulfation by sulfotransferases allowed us to map glycan epitopes in biological samples. This was illustrated using mouse embryo and rat organ tissue sections labeled with the following carbohydrate sulfotransferases: CHST3, CHST15, HS3ST1, CHST4 and CHST10.

Direct MALDI-MS Analysis of Cardiolipin from Rat Organs Sections

Cardiolipins (CL) are mitochondria specific lipids. They play a critical role in ATP synthesis mediated by oxidative phosphorylation. Abnormal CL distribution is associated with several disease states. MALDI-MS and MALDI-MS/MS were used to demonstrate in situ analysis and characterization of CL from tissue sections of organs containing high concentrations of mitochondria. Once the experimental parameters were established, a survey of CL distribution in heart, liver, kidney, leg muscle, and testis was undertaken. The major CL specie in the heart muscle, leg muscle, liver, and kidney is the (18:2)(4) CL, while liver and kidney also contain a minor specie, (18:2)(3)/(18:1) CL. The major CL specie in testis is the (16:0)(4) CL. The CL species distribution in various organs appeared to be in agreement with prior reports. Overall, proper matrix selection, tissue section handling, instrument tuning, and the inclusion of cesium ion in matrix ensured successful in situ MALDI-MS and MALDI-MS/MS analysis of CL. Upon modification and standardization, this method could be streamlined for rapid pathological diagnosis with short turnaround time in clinical settings.

Neointimal growth can be influenced by local adventitial gene manipulation via a needle injection catheter

Revascularization by percutaneous transluminal coronary angioplasty is limited in the long-term by restenosis, which is luminal renarrowing in the first 6 months after the procedure. Smooth muscle cell proliferation is thought to be an important factor in restenosis; this leads to neointima formation and arterial lumen narrowing. Local therapy delivered perivascularly may have an effect on events in the neointima and reduce restenosis. The effect of delivering expression vector plasmids for senescent cell-derived inhibitor SDI-1, which regulates cell proliferation, and its antisense, into the perivascular tissue of injured arteries was investigated in a porcine arterial injury model using a needle injection catheter. Transfection efficiency, biological effect and plasmid dissemination were evaluated in arterial and organ tissue sections between 2 days and 4 months. A limited number of adventitial, medial and neointimal cells were transfected up to 4 months. sdi gene transfer did not result in a change in neointima. Transfer of antisense sdi resulted in an increase in neointima after 3 weeks. No DNA plasmid was detected in control tissues. Liposomally-mediated adventitial local gene delivery is feasible and safe using the needle injection catheter in a porcine model. A limited number of cells was transfected, with expression of transfected genes up to 4 months after delivery. A transient biological effect with increased neointima was observed after delivery of the antisense sdi gene.

Differentiation of Hog Cholera and Bovine Virus Diarrhoea Viruses in Pigs using Monoclonal Antibodies

Monoclonal antibodies against hog cholera and bovine viral diarrhoea viruses were assayed on organ tissue sections of experimentally infected animals. The animals had been infected simultaneously with both viruses. The antibodies were tested using an indirect immunofluorescence test and an indirect enzyme immunoassay with a biotin/streptavidin/peroxidase detection system. A polyclonal hyperimmune serum was used as a control in direct immunofluorescence tests. Both techniques based on monoclonal antibodies were more sensitive and more specific than the conventional test, the enzyme immunoassay being more sensitive than the immunofluorescence test. Small amounts of BVD viral antigen were demonstrable with monoclonal antibodies in most organ tissues.