In situ hybridization

cDNAs of rat GR (~0.4 kb) and rat SERT (~0.7 kb) in pGEM-3Zf vectors with T7 RNA polymerase and DIG RNA labeling mix (Roche, Branford, CT, USA). The amount of the transcript was estimated by comparison to DIG-labeled control RNA (Roche, Branford, CT, USA) in Dot-blot assays on nylon membranes (Roche, Branford, CT, USA). The ISH protocol is similar to the one from Cold Spring Harbor Laboratory with minor modifications (Javelle et al. 2011). In brief, slides were fixed with 4% (w/v) paraformaldehyde followed by 0.05% (w/v) proteinase K treatment for 5 min at 21oC and acetylation with acetic anhydride. Lipids were extracted by washing through an ethanol series: 50, 70, 95 and 100% (vols). Pre-hybridized sections were incubated with 200 µl hybridization solution per slide containing 200 ng DIG-labeled probes at 55oC overnight. At the next morning, the sections were washed extensively and treated with RNase A, then proceed to block in 4% goat serum for one hour. The anti-DIG-alkaline phosphatase antibody (1:200 dilution, Roche, Branford, CT, USA) was applied to sections at 4oC overnight. The color was developed by NBT/BCIP substrates (Roche, Branford, CT, USA ) with refreshments twice a day for 3~5…

molecular genetic tests for NF1: Comprehensive multistep mutation detection, Genomic DNA sequence analysis, Deletion/ duplication analysis, and Cytogenetic analysis. The comprehensive multistep mutation detection protocol has been effective in detecting 95% of individuals with a pathogenic NF1 variant while Genomic sequencing, deletion/duplication analysis, and Cytogenetic analysis have only been able to detect a defect in the NF1 gene in 61%, 5%, and less than 1% of individuals. The most…

nanoparticles in samples were visualised. Bio-distribution of nanoparticles in samples was also observed by the confocal fluorescence microscopy, hyperspectral fluorescence microscopy and transmission electron microscopy imaging for confirmation purpose1. Bioluminescence observation suggested a significant proportion of samples were successfully labelled with nanodots and they can be clearly visualised in all three different experimental conditions: in vitro (labelled spermatozoa and oocytes),…

A quote to highlight the misuse of medical ethics is “TeLinde disagreed— he believed carcinoma in situ was simply an early stage of invasive carcinoma that, if left untreated, eventually became deadly. So he treated it aggressively, often removing the cervix, uterus, and most of the vagina” (Skloot 165). This was unethical because there was no proven way to tell if this was the best treatment method, he did it just because he thought it was best. This is an example of a doctor using his own…

Most gynecologic oncologists use the FIGO (International Federation of Gynecologists and Obstetricians) classification. The disease is divided into five stages, with further divisions in each stage. A carcinoma in situ is Stage 0. In Stage I, the cancer is confined to the cervix. When the disease spreads beyond the cervix but not to the pelvic sidewall, or involves the vagina but not the lower third, this is considered Stage II. Stage III carcinoma spreads to the pelvic sidewall, involves the…

CHAPTER 2.LITERATURE REVIEW 2.1. Definition The cervix is the lower part of the uterus (womb). It is sometimes called the uterine cervix. It connects the body of uterus to the vagina. The part of the cervix inside the cervix canal is called endocervix. The part on the outside is called the ectocervix. Most cervical cancers start where these two parts meet. There are two main types, squamous cell carcinoma and adenocarcinoma. Occasionally, mixed carcinomas,…

This island has to be saved for its trees, it has to be saved for its animals it is a part of a reserve forest, it belongs to a project to save tigers, which is paid for by people from all around the world….who love animals so much that they are willing to kill us for them…this whole world has become a place for animals, and our fault, our crime was that we were just human beings, trying to live as human beings always have, from the water and the soil. No human being…

Animals are born with natural instincts that kick in at a very young age. If an animal is raised in a zoo it may not know what to do when released into the wild. These animals may not be able to survive. So which is best for the animals, the wild or the zoo? Honestly, I believe animals should live in the wild. When an animal is born in captivity their natural instincts may not kick in. These animals are raised without the fear of predators, the instinct to hunt, or even to be on their own. Once…

Captive breeding and reintroduction programs are important tools for the establishment of threatened animal populations, but experimental approaches require to compare the outcomes of these programs that will increase post-release survival. The Kaiser’s Mountain Newt, Neurergus kaiseri is listed as a vulnerable by IUCN criteria, to aid about the ecology and the conservation of this species, we carried out a trial release of 20 newts reared in captivity on 4 occasions during the spring and summer…

mention, they participate in internships at animals sanctuaries, animals camps, and veterinary hospitals. When handling these exotic animals, zookeepers do it with care as they were trained to so. These zookeepers maintain the animals’ habitat and also engage them in activities. Care has improved a lot over the last few decades mostly because of the improvement in education that zookeepers attain. They now understand that animals need to be kept busy and active in order to prevent boredom and…