Effects of probiotic supplements on insulin resistance in gestational diabetes mellitus: A double‐blind randomized controlled trial

Abstract Aims/Introduction To evaluate the effect of probiotic supplements on insulin resistance in pregnant women with diet‐controlled gestational diabetes mellitus. Materials and Methods A randomized, double‐blind, placebo‐controlled trial was carried out between June 2016 and February 2017. Pregnant women with diet‐controlled gestational diabetes mellitus were enrolled in the study at 24–28 weeks‐of‐gestation and randomized to receive either probiotic supplements containing Bifidobacterium and Lactobacillus or a placebo daily for four consecutive weeks. Primary outcomes were mean differences in insulin resistance (homeostatic model assessment for insulin resistance), fasting insulin and fasting plasma glucose between the two groups. Secondary outcomes were changes in maternal weight after the intervention. Results Data from 28 patients in the probiotic group and 29 in the placebo group were analyzed. The changes in metabolic parameters after randomization showed significant improvement in glucose metabolism in the probiotic group compared with the placebo group, including fasting plasma glucose (0.68 ± 5.88 vs 4.620 ± 7.78 mg/dL, mean difference −3.94 mg/dL, 95% confidence interval −7.62, −0.27, P = 0.034), fasting plasma insulin (1.11 ± 1.71 vs 3.77 ± 1.70 mIU/L, mean difference −2.67 mIU/L, 95% confidence interval −3.57, −1.76, P = 0.001) and homeostatic model assessment for insulin resistance (0.25 ± 0.37 vs 0.89 ± 0.46, mean difference −0.63, 95% confidence interval −0.86, −0.41, P = 0.001). Weight gain during randomization was similar between the two groups. Conclusions Four weeks of probiotic supplements in women with diet‐controlled gestational diabetes in the late second and early third trimester lowered fasting glucose and increased insulin sensitivity. Probiotic supplements may be considered as an adjunct treatment for glycemic control in these patients

Parental Awareness, Knowledge, and Attitudes Regarding Current and Future Newborn Bloodspot Screening: The First Report from Thailand

Newborn screening (NBS) is a public health service that is used to screen for treatable conditions in many countries, including Thailand. Several reports have revealed low levels of parental awareness and knowledge about NBS. Because of limited data on parental perspectives toward NBS in Asia and the differences in socio-cultural and economic contexts between Western and Asian countries, we conducted a study to explore parental perspectives on NBS in Thailand. A Thai questionnaire to assess awareness, knowledge, and attitudes regarding NBS was constructed. The final questionnaire was distributed to pregnant women, with or without their spouses, and to parents of children aged up to one year who visited the study sites in 2022. A total of 717 participants were enrolled. Up to 60% of parents were identified as having good awareness, which was significantly associated with gender, age, and occupation. Only 10% of parents were classified as having good knowledge relative to their education level and occupation. Providing appropriate NBS education should be initiated during antenatal care, focusing on both parents. This study noted a positive attitude toward expanded NBS for treatable inborn metabolic diseases, incurable disorders, and adult-onset diseases. However, modernized NBS should be holistically evaluated by multiple stakeholders in each country because of different socio-cultural and economic contexts

Lupeol and stigmasterol suppress tumor angiogenesis and inhibit cholangiocarcinoma growth in mice via downregulation of tumor necrosis factor-α

<div><p>Lupeol and stigmasterol, major phytosterols in various herbal plants, possess anti-inflammatory activities and have been proposed as candidates for anti-cancer agents, but their molecular mechanisms are still unclear. Here, we investigated the effects of lupeol and stigmasterol on tumor and endothelial cells in vitro and their anti-cancer activities in vivo. Our results demonstrated that lupeol and stigmasterol suppressed cell viability, migration, and morphogenesis of human umbilical vein endothelial cells (HUVECs) but not cholangiocarcinoma (CCA) cells. Expression analyses showed that the treatment of both compounds significantly reduced the transcript level of tumor necrosis factor-α (TNF-α), and Western blot analyses further revealed a decrease in downstream effector levels of VEGFR-2 signaling, including phosphorylated forms of Src, Akt, PCL, and FAK, which were rescued by TNF-α treatment. In vivo, lupeol and stigmasterol disrupted tumor angiogenesis and reduced the growth of CCA tumor xenografts. Immunohistochemical analyses confirmed a decrease in CD31-positive vessel content and macrophage recruitment upon treatment. These findings indicate that lupeol and stigmasterol effectively target tumor endothelial cells and suppress CCA tumor growth by their anti-inflammatory activities and are attractive candidates for anti-cancer treatment of CCA tumors.</p></div

Treatment of lupeol and stigmasterol reduced tumor angiogenesis, tumor growth, and macrophage recruitment in cholangiocarcinoma xenograft models.

<p>(A) KKU-M213 and (B) RMCCA-1 tumors weighed less in the lupeol or stigmasterol, or combination treatment groups compared with control. Data, as mean tumor weight ± SD; *P value < 0.05 (n = 4–5). (C) CD31 staining of compound-treated KKU-M213 tumors; scale bar, 200 μm. (D) Quantification of CD31-positive areas. Data, mean percentage of the CD31-positive area ± SD; *P < 0.003 (n = 4–5). (E) H&E, Masson’s trichrome, and (F) F4/80 staining of KKU-M213 tumors. Scale bar, 200 μm. L, lupeol; S; stigmasterol.</p

Lupeol and stigmasterol did not exhibit significant toxic effects in mice.

<p>(A) Treatment timeline for the toxicity studies. (B) No significant difference in weight change was observed between the control and treatment groups. Data presented as percent weight mean change ± S.D. (n = 5). (C) H&E staining of the livers showed no difference between the control and treatment groups. (D) Sera were also collected from mice for blood chemistry analyses. No significant differences were observed in ALT, AST, RBC, Hemoglobin, WBC, Hematocrit using one-way ANOVA. Group 1, control; Group 2, lupeol (1 mg/kg); Group 3, lupeol (10 mg/kg); Group 4, stigmasterol (1 mg/kg); Group 5, stigmasterol (10 mg/kg); Group 6, lupeol + stigmasterol (1 mg/kg); Group 7, lupeol + stigmasterol (10 mg/kg).</p

Lupeol and stigmasterol inhibited HUVEC migration and capillary network formation in vitro.

<p>(A) Lupeol and stigmasterol inhibited HUVEC migration. The effects of the compounds on HUVEC migration abilities were determined using 2-dimensional wound healing assays. HUVECs were seeded at 100% confluency and scratched at the middle of each well, then each compound was added to the culture at 5 μM concentration. Scale bar, 50 μm. (B) Lupeol and stigmasterol also significantly disrupted HUVEC network formation. Endothelial network formation assays were performed by seeding HUVECs between two type I collagen gel layers and cultured for 4 days. Scale bar, 200 μm.</p

Lupeol, β-sitosterol, and stigmasterol exhibited cytotoxic effects on HUVECs.

<p>Structures and cell viability of lupeol (A), β-sitosterol (B), and stigmasterol (C). Cell viability of HUVECs, which were exposed for 48 hours to different doses of lupeol, β-sitosterol, and stigmasterol, as determined by MTT assays. Means ± S.D. *P value < 0.05 vs. control. n = 3. (D) The IC₅₀ values of bevacizumab, lupeol, β-sitosterol, and stigmasterol in HUVECs. Means ± S.D.</p

Lupeol and stigmasterol downregulated TNF-α and inhibited VEGF signaling.

<p>The expression of inflammatory cytokines and components of the VEGF pathway was examined by qRT-PCR in HUVECs treated with the compounds at 5 μM. (A-C) Lupeol and stigmasterol suppressed expression levels of TNF-α but showed no impact on IL-6 and CXCL-8. (D-F) Expression of VEGFR-2, but not VEGF-A or VEGFR-1, significantly decreased upon lupeol and stigmasterol treatment. (G) Western blotting showed a decrease in the p-FAK level at both concentrations and a slight decrease in levels of p-Src, p-PCL, p-Akt only at low concentration. L, lupeol; S; stigmasterol. (H) Densitometry calculations for TNF-αWestern blot data in (G) using ImageJ normalized to beta-actin. (I) ELISA for secreted TNF-alpha in the conditioned media from HUVECs treated with lupeol or stigmasterol or combination. Means ± S.D. *P value < 0.05 vs. control. n = 3. BV, bevacizumab; L, lupeol; S, stigmasterol; (1), 1 μM; (5), 5 μM.</p