Isolation and Characterization of Neural Crest-Derived Stem Cells from Dental Pulp of Neonatal Mice

Dental pulp stem cells (DPSCs) are shown to reside within the tooth and play an important role in dentin regeneration. DPSCs were first isolated and characterized from human teeth and most studies have focused on using this adult stem cell for clinical applications. However, mouse DPSCs have not been well characterized and their origin(s) have not yet been elucidated. Herein we examined if murine DPSCs are neural crest derived and determined their in vitro and in vivo capacity. DPSCs from neonatal murine tooth pulp expressed embryonic stem cell and neural crest related genes, but lacked expression of mesodermal genes. Cells isolated from the Wnt1-Cre/R26R-LacZ model, a reporter of neural crest-derived tissues, indicated that DPSCs were Wnt1-marked and therefore of neural crest origin. Clonal DPSCs showed multi-differentiation in neural crest lineage for odontoblasts, chondrocytes, adipocytes, neurons, and smooth muscles. Following in vivo subcutaneous transplantation with hydroxyapatite/tricalcium phosphate, based on tissue/cell morphology and specific antibody staining, the clones differentiated into odontoblast-like cells and produced dentin-like structure. Conversely, bone marrow stromal cells (BMSCs) gave rise to osteoblast-like cells and generated bone-like structure. Interestingly, the capillary distribution in the DPSC transplants showed close proximity to odontoblasts whereas in the BMSC transplants bone condensations were distant to capillaries resembling dentinogenesis in the former vs. osteogenesis in the latter. Thus we demonstrate the existence of neural crest-derived DPSCs with differentiation capacity into cranial mesenchymal tissues and other neural crest-derived tissues. In turn, DPSCs hold promise as a source for regenerating cranial mesenchyme and other neural crest derived tissues

Optimising blood glucose control with portioned meal box in type 2 diabetes mellitus patients: a randomised control trial

BackgroundThe impact of dietary factors on glycaemic control in type 2 diabetes mellitus (T2DM) is well established. However, the effectiveness of transforming portion control into a practical innovation for glycaemic control in T2DM has not yet been established for counselling in nutrition. The aim of this study was to compare the effect of general counselling in nutrition (GCN) and a portioned meal box (PMB) on fasting blood glucose, glycated haemoglobin (HbA1c) and body composition.MethodsA randomised, parallel intervention trial was conducted over 12 weeks, with GCN: carbohydrate portion control concept by using food exchange lists (n = 25) and PMB: portioned meal box was set by energy requirements (n = 25).ResultsBoth GCN and PMB demonstrated reductions in HbA1c levels at the 6th and 12th weeks compared to baseline. However, no significant difference in HbA1c was observed between GCN and PMB at either the 6th or 12th week. Using PMB at least four times a week significantly decreased HbA1c during the intervention period (p = 0.021 and p < 0.001 for weeks 6 and 12 when compared with baseline, respectively). Changes in body composition were observed: body weight decrease in PMB only, body fat decrease and constant muscle mass in both groups. Both methods tended to relieve hunger and increased satiety in both groups. The satisfaction evaluation showed that participants preferred to use PMB over GCN (p = 0.001). Additionally, participants consumed less energy, carbohydrate and fat in PMB (p = 0.001, p = 0.019, and p = 0.001, respectively) and less energy and fat in GCN (p = 0.006 and p = 0.001, respectively).ConclusionA better diet, either through GCN or PMB, can play an important role in improving dietary intake compliance and controlling blood glucose

Deposition of Triglyceride on Soft Contact Lenses from Lipid-Containing Artificial Tears

Objective: To determine the amount of triglyceride deposition from a lipid-containing artificial tear eye drop (Endura® ) on three different types of contact lenses after one day and one month of simulated use. Methods: Simulated use of artificial tears was performed in vitro on three different types of contact lenses, including: high water content (HW), low water content (LW), and silicone hydrogel (SI) contact lenses. To simulate one day of use, contact lenses were incubated in artificial tear solution for 16 hours. To simulate a one month of use, they underwent repeated cycles of 16-hour incubation in artificial tear solution, lens cleaning, and 8-hour storage in a multipurpose solution daily for 30 days. Triglyceride deposited was extracted and determined. Results: After one day of simulated use, amount of deposited triglyceride was significantly different among 3 types of contact lenses with the highest deposition in SI followed by LW and HW contact lenses; the corresponding values (mean ± SD) were 3.79 ± 0.35, 0.84 ± 0.27, and 0.26 ± 0.17 μg/lens, respectively. Only between SI and HW was it found to be statistically different. After one month of simulated use, deposition on SI lenses was slightly increased (6.56 ± 1.10 μg/lens) with that on LW and HW lenses remaining low (0.10 ± 0.12 and 0.55 ± 0.34 μg/lens, respectively). Conclusion: Triglyceride from lipid-containing artificial tears can absorb into contact lenses, particularly those made of silicone hydrogel, most notably after long periods of use. However, levels of triglyceride deposition are relatively low, when compared to lipid deposition from normal tear films during regular use

Isolation, Characterization, and Transplantation of Cardiac Endothelial Cells

Isolation and ex vivo expansion of cardiac endothelial cells have been a recurrent challenge due to difficulties in isolation, cell heterogeneity, lack of specific markers to identify myocardial endothelial cells, and inadequate conditions to maintain long-term cultures. Herein, we developed a method for isolation, characterization, and expansion of cardiac endothelial cells applicable to study endothelial cell biology and clinical applications such as neoangiogenesis. First, we dissociated the cells from murine heart by mechanical disaggregation and enzymatic digestion. Then, we used flow cytometry coupled with specific markers to isolate endothelial cells from murine hearts. CD45+ cells were gated out to eliminate the hematopoietic cells. CD31+/Sca-1+ cells were isolated as endothelial cells. Cells isolated from atrium grew faster than those from ventricle. Cardiac endothelial cells maintain endothelial cell function such as vascular tube formation and acetylated-LDL uptake in vitro. Finally, cardiac endothelial cells formed microvessels in dorsal matrigel plug and engrafted in cardiac microvessels following intravenous and intra-arterial injections. In conclusion, our multicolor flow cytometry method is an effective method to analyze and purify endothelial cells from murine heart, which in turn can be ex vivo expanded to study the biology of endothelial cells or for clinical applications such as therapeutic angiogenesis

Effect of Thunbergia laurifolia Herbal Tea on Glucose Homeostasis in Healthy Volunteers: A Single-Arm Phase I Study

Background. Thunbergia laurifolia (TL) is a commonly used herbal medicine in Thailand and in other Asian countries. TL has been approved as a Thai traditional medicine for detoxifying poisons, and the list of possible adverse effects includes hypoglycemia. TL showed hypoglycemic effect in animals possibly due to antioxidant effect and beta-cell preservation. However, the safety of TL herbal tea and its effects on glucose homeostasis have never been investigated in humans. Methods. Twenty healthy volunteers (10 men and 10 women) drank TL herbal tea 3 times/day for 2 weeks. Ten subjects took TL herbal tea 9 grams daily. After the safety of TL herbal tea was established, 10 more subjects took TL 12 grams daily. Clinical and biochemical tests were assessed at baseline and at 2 weeks. Results. Mean age was 34.9 ± 10.2 years, and mean body mass index was 27.5 ± 5.8 kg/m2. Baseline and posttreatment plasma concentrations were as follows: fasting plasma glucose (89 ± 6 vs. 89 ± 7 mg/dL), fructosamine (213 ± 32 vs. 212 ± 33 μmol/L), fasting insulin (8.8 [IQR: 5.9–18.4] vs. 10.4 [IQR: 7.4–15.2] μU/mL), HOMA-B (101.6 [IQR: 82.3–189.8] vs. 120.4 [IQR: 93.2–153.2]), and HOMA-IR (1.1 [IQR: 0.8–2.3] vs. 1.4 [IQR: 0.9–2.0]), all respectively. There were no significant changes in these parameters, including body weight, blood pressure, lipid profile, and C-reactive protein. No serious adverse events were observed during the study period. Conclusions. TL herbal tea at doses of 9 and 12 grams daily had good tolerability without any significant adverse effects on fasting plasma glucose level or other glucose homeostasis parameters measured

Autoverification improved process efficiency, reduced staff workload, and enhanced staff satisfaction using a critical path for result validation

Objective: Continuous process improvements enhance accuracy and productivity in a clinical laboratory setting. This study aimed to investigate the accuracy and efficiency of a new autoverification (AV) system designed to improve the consistency and uniformity of reported laboratory test results. Methods: Limit checks, delta checks, and consistency checks were established, and then retrospective data from 500 requested tests were used to evaluate the accuracy of AV rules compared to manual verification, which was performed by five experienced medical technologists. Efficiency was evaluated by comparing turnaround time (TAT), error rates, workload, and staff satisfaction between before and after AV implementation. Results: AV had 100% sensitivity, 77.6% specificity, and a 22% false-positive rate. The AV passing rate was 95%, 85%, 42%, and 39% for chemistry, coagulation, microscopy, and hematology, respectively. The overall passing rate was 65%. After implementation, the mean overall TAT decreased from 54.2±26.6 to 52.4±24.2 min (p<0.001). However, TAT during peak hours increased (p<0.05). Incident reports decreased 8-fold (p<0.05), net workload decreased by 0.76 full-time equivalent, and overall staff satisfaction increased (p<0.001). Conclusion: Our laboratory’s new AV system demonstrated an overall passing rate of 65% with decreases in TAT, incident reports, and workload, and an increase in staff satisfaction

Maternal Vitamin D Status and Its Related Factors in Pregnant Women in Bangkok, Thailand

<p>Dataset from manuscript submitted to PLOS ONE entitled</p> <p>"Maternal Vitamin D Status and Its Related Factors in Pregnant Women in Bangkok, Thailand" by </p> <p>Busadee Pratumvinit, Preechaya Wongkrajang,Tuangsit Wataganara, Sithikan Hanyongyuth, Akarin Nimmannit, Somruedee Chatsiricharoenkul, Kotchamol Manonukul, Kanit Reesukumal</p

Scatter plot showing the relationship between maternal intact parathyroid hormone and 25-hydroxyvitamin D level.

<p>The maternal plasma level of 25-hydroxyvitamin D [25(OH)D]showed a weakly negative correlation with the maternal plasma level of intact parathyroid hormone (iPTH), n = 147. Linear regression equation: iPTH = –0.0193[25(OH)D] + 4.3884. (Pearson’s <i>r</i> = –0.29, <i>P</i><0.01)</p