Notch signaling promotes intestinal crypt fission in the infant rat

BACKGROUND Growth of the small intestine in the infant rat is promoted by crypt fission and later by increased crypt cell proliferation. Notch signaling could promote crypt fission. Hes-1 is a Notch target gene. AIM We assessed the effect of Notch signaling on intestinal crypt fission and on growth of the intestine in the infant rat. METHODS Hes-1 expression was determined in the small intestine of litters of Hooded Wistar rats aged between 3 and 72 days. Hes-1 RNA expression was measured by quantitative RT-PCR. Four groups of rats (n = 8 or 9) were injected daily, ip, either with vehicle or with the Notch inhibitor DAPT at doses of 3, 10, and 30 mg/kg, from days 9 to 13 of life, and killed on day 14. A microdissection technique was used to measure crypt fission, mitotic count, and apoptotic count. Data were analyzed by ANOVA and by use of Dunnett’s F test. RESULTS Hes-1 expression and crypt fission peaked on day 14. DAPT reduced Hes-1 immunostaining in proportion to dose. DAPT reduced villous area to 72 % (p < 0.01), 53 % (p < 0.001), and 38 % (p < 0.001) of control values for 3, 10 and 30 mg/kg doses, respectively, and reduced crypt fission to 53 % (p < 0.001) and 38 % (p < 0.001) of control values, respectively, for 10 and 30 mg/kg doses. Crypt mitotic count was not affected by any DAPT dose. DAPT at 10 and 30 mg/kg significantly increased apoptosis in crypts, by 6.5 and 4.8-fold, respectively. CONCLUSIONS We conclude that Notch signaling promotes crypt fission and growth of the intestine by maintaining low apoptosis of crypt cells.Adrian G. Cummins, Joshua A. Woenig, Rino P. Donato, Simon J. Proctor, Gordon S. Howarth, Phulwinder K. Grove

Analysis of Wnt signalling dynamics during colon crypt development in 3D culture

Many systems biology studies lack context-relevant data and as a consequence the predictive capabilities can be limited in developing targeted cancer therapeutics. Production of colon crypt in vitro is ideal for studying colon systems biology. This report presents the first production of, to our knowledge, physiologically-shaped, functional colon crypts in vitro (i.e. single crypts with cells expressing Mucin 2 and Chromogranin A). Time-lapsed monitoring of crypt formation revealed an increased frequency of single-crypt formation in the absence of noggin. Using quantitative 3D immunofluorescence of β-catenin and E-cadherin, spatial-temporal dynamics of these proteins in normal colon crypt cells stimulated with Wnt3A or inhibited by cycloheximide has been measured. Colon adenoma cultures established from APC(min/+) mouse have developmental differences and β-catenin spatial localization compared to normal crypts. Quantitative data describing the effects of signalling pathways and proteins dynamics for both normal and adenomatous colon crypts is now within reach to inform a systems approach to colon crypt biology

Chemical composition and health benefits of coconut oil: an overview

Coconut oil is an integral part of Sri Lankan and many South Asian diets. Initially, coconut oil was classified along with saturated fatty acid food items and criticized for its negative impact on health. However, research studies have shown that coconut oil is a rich source of medium-chain fatty acids. Thus, this has opened new prospects for its use in many fields. Beyond its usage in cooking, coconut oil has attracted attention due to its hypocholesterolemic, anticancer, antihepatosteatotic, antidiabetic, antioxidant, anti-inflammatory, antimicrobial and skin moisturizing properties. Despite all the health benefits, consumption of coconut oil is still underrated due to a lack of supportive scientific evidence. Even though studies done in Asian countries claim a favorable impact on cardiac health and serum lipid profile, the limitations in the number of studies conducted among Western countries impede the endorsement of the real value of coconut oil. Hence, long-term extensive studies with proper methodologies are suggested to clear all the controversies and misconceptions of coconut oil consumption. This review discusses the composition and functional properties of coconut oils extracted using various processing methods