From lessons on the long-term effects of the preimplantation environment on later health to a "modified ART-DOHaD" animal model.

Background: At its earliest stages, mammalian embryonic development is apparently simple but vulnerable. The environment during the preimplantation period, which only lasts a couple of days, has been implicated in adult health, extending to such early stages the concept of the developmental origin of health and disease (DOHaD). Methods: In this review, we first provide a brief history of assisted reproductive technology (ART) focusing on in vitro culture and its outcomes during subsequent development mainly in mice and humans. Further, we introduce the "MEM mouse," a novel type 2 diabetes mouse model generated by in vitro culture of preimplantation embryos in alpha minimum essential medium (αMEM). Main findings: The association between ART and its long-term effects has been carefully examined for its application in human infertility treatment. The "MEM mouse" develops steatohepatitis and kidney disease with diabetes into adulthood. Conclusion: The close association between the environment of preimplantation and health in postnatal life is being clarified. The approach by which severe mouse phenotypes are successfully induced by manipulating the environment of preimplantation embryos could provide new chronic disease animal models, which we call "modified ART-DOHaD" animal models. This will also offer insights into the mechanisms underlying their long-term effects.

Heterogeneity of nucleolar morphology in four-cell mouse embryos after IVF: association with developmental potential.

In mammals, around fertilization, the nucleolus of embryos transforms into the nucleolus precursor bodies (NPBs), which continue to mature until the blastocyst stage, leading to distinct morphological changes. In our study, we observed two types of nucleolar morphology in mouse in vitro fertilized embryos at the four-cell stage, which we refer to single nucleolus (SN) and multiple nucleoli (MN). To visualize nucleolar morphology, four-cell embryos were immunostained with anti-NOPP140 antibody. These embryos were categorized into five types based on the number of blastomeres carrying SN: SN4/MN0, SN3/MN1, SN2/MN2, SN1/MN3, and SN0/MN4, with percentages of 13, 27, 21, 23 and 9, respectively. Next, using a light microscope, we divided the four-cell in vitro fertilized embryos without fixation into two groups: those with at least two blastomeres displaying SN (SN embryos) and those without (MN embryos). Notably, significantly more SN embryos developed into blastocysts and offspring at 18.5 dpc compared with MN embryos. Furthermore, SN embryos displayed a higher NANOG-positive cell number at the blastocyst stage, significantly lower body and placental weights, resulting in a higher fetal/placental ratio. These findings suggest a close association between nucleolar state at the four-cell stage and subsequent developmental potential.

Clonal Blumea lacera (Burm. f.) DC. ameliorates diabetic conditions by modulating carbohydrate and lipid hydrolases: a combine in vivo experimental and chemico-biological interaction study.

Blumea lacera (Burm. f.) DC. is an aromatic annual herb that has traditionally been used to treat or protect against diabetes. Although it has infallible uses, its supply is limited due to its short lifespan. In this study, we aim to investigate the anti-diabetic potential of its micropropagated plants in type 2 diabetic mammalian (mouse) model and further expand the molecular mechanistic understanding of its activity. The water extract of the micropropagated plants was tested in mice with streptozotocin-induced diabetes. The extract effectively suppressed glucose levels prevented weight loss, and improved dyslipidemia in mice. Additionally, it improved liver injury as well as all investigated toxicity indicators, including serum glutamate-pyruvate transaminase, serum glutamic oxaloacetic transaminase, and serum anti-inflammatory marker C-reactive protein. The intramolecular interaction study revealed that the innate polyphenolic constituents of this plant more profoundly inhibited α-amylase, α-glucosidase, and lipase compared to the standard. The prolific bioactive compounds of the micropropagated plant could be attributed to these superior anti-diabetic effects, presumably via an elaborate inhibition of carbohydrate and lipid hydrolyzing enzymes. Thus, the obtained results provide solid experimental proof of the year-round utility of micropropagated plants as a standard source plant material of Blumea lacera (Burm. f.) DC. for drug research and therapeutic production.

Determination of in vitro antioxidant activity and in vivo antineoplastic effects against Ehrlich ascites carcinoma of methanolic extract of Sphagneticola calendulacea (L.) Pruski.

BACKGROUND AND AIM: Sphagneticola calendulacea (L.) Pruski (S. calendulacea) is a well-known medicinal plant that is widely used in traditional medicine. This study was conducted to evaluate the antioxidant and antineoplastic properties of S. calendulacea leaf. EXPERIMENTAL PROCEDURE: Methanol was used as a solvent to prepare leaf extract from the dried leaf of S. calendulacea (MESL). We have evaluated the total phenolic and flavonoid contents of MESL, and using five different assays; we have also evaluated the antioxidant property of MESL. In vivo antineoplastic activity of MESL against Ehrlich ascites carcinoma (EAC) cells in experimental mice was also explored. RESULTS: The phytochemical analysis of MESL exhibited the total phenolic, and total flavonoid contents in the dry extracts were 32.86 mg/gm of gallic acid equivalent and 127.26 mg/gm of catechin equivalent, respectively. Moreover, MESL also showed promising scavenging activity in all assays (IC50 value ≥70.5 µg/mL). In the in vivo antineoplastic assay, MESL showed increased EAC cell death at the doses of 50 mg/kg and 100 mg/kg body weight. MESL administration also induced apoptosis of EAC cells, significantly inhibited EAC cell growth, and increased the life span of EAC cell-bearing mice compared to EAC cell-bearing control mice. CONCLUSIONS: All findings of this study suggest potential antioxidant and antineoplastic properties of MESL.

Phytochemical profiling of Blumea laciniata (Roxb.) DC. and its phytopharmaceutical potential against diabetic, obesity, and Alzheimer's.

Blumea laciniata (Roxb.) DC. is a folk medicinal annual herb of the Asteraceae family that grows in South and Southeast Asia. In order to evaluate its phytopharmaceutical potential against diabetic, obesity, and Alzheimer's, a comprehensive phytochemical profile, in vitro and in silico enzyme inhibitory activity against α-amylase, α-glucosidase, lipase, cholinesterases, and tyrosinase along with in vitro antioxidant activity were performed. Additionally, in vivo antidiabetic activity and acute toxicity were also evaluated. The total phenolic content in various organs follows the following order: old leaf > flower bud > young leaf > flower > young stem > old stem > root, while total flavonoids followed the order: flower bud > old leaf > young leaf > flower > young stem > old stem > root. The identified phenolic compounds are 3,4-dihydroxybenzoic acid, caffeic acid, vanillic acid, p-coumaric acid, syringic acid, rosmarinic acid, trans-cinnamic acid, catechin, catechol, (-) epicatechin, rutin, quercetin, myricetin, and kaempferol, which are also expressed differently in various organs. Solvent extracts demonstrated strong antioxidant activity as well as varying levels of inhibition against the enzymes tested, with strong inhibitory activity against α-amylase, α-glucosidase, and lipase. Thirteen phenolic compounds displayed strong binding affinity in silico against studied enzymes, thus documented as bioactive. Furthermore, solvent extracts significantly suppressed blood glucose levels in mice with induced diabetes and extracts were not acutely toxic. The results suggest that Blumea laciniata (Roxb.) DC. could be a potential candidate for developing new phytopharmaceuticals and bioactive ingredients.