A prospective case series in regenerative endodontics: The effective use of diluted antibiotic hydrogels in endodontic regeneration procedures.

Objective: to investigate the effectiveness of diluted antibiotic hydrogels in endodontic regeneration procedures. Materials and Methods: One conventional treatment (calcium hydroxide) and two experimental treatments containing 1 mg/mL antibiotic in gel (MC-TAP and MC-DAP) were prepared for this study. The effect of these medicaments on the proliferation and differentiation potential of dental pulp stem cells (DPSCs) was examined before their clinical use, using WST and ALP assays. Twenty-two teeth diagnosed with necrotic immature roots were treated with calcium hydroxide, MC-TAP or MC-DAP using the endodontic regeneration protocol as described by the American Association of Endodontists (AAE). Teeth were examined after application of the medicament, at the time of regeneration and after 12-24 months. Results: All treated teeth showed resolution of signs and symptoms shortly after treatment with the different medicaments. However, signs and symptoms returned a few months after initial treatment in clinical cases treated with MC-DAP. All successfully treated teeth examined after 12-24 months were free of signs and symptoms, but vitality was not achieved. Conclusion: Low concentration antibiotic gel containing triple antibiotic paste (MC-TAP) can be successfully used for endodontic regeneration procedures.

Intracytoplasmic sperm injection is not superior to conventional IVF in couples with non-male factor infertility and preimplantation genetic testing for aneuploidies (PGT-A).

STUDY QUESTION: Does the insemination method impact the euploidy outcome in couples with non-male factor infertility? SUMMARY ANSWER: Conventional IVF can be applied in cycles with preimplantation genetic testing for aneuploidies (PGT-A), as both IVF and ICSI generate equal numbers of euploid blastocysts. WHAT IS KNOWN ALREADY: Ever since its introduction, the popularity of ICSI has increased tremendously, even in couples with non-male factor infertility. The use of conventional IVF is a contraindication for couples undergoing PGT to ensure monospermic fertilisation and to eliminate potential paternal contamination from extraneous sperm attached to the zona pellucida. Despite this, it has recently been shown that sperm DNA fails to amplify under the conditions used for trophectoderm biopsy samples. STUDY DESIGN, SIZE, DURATION: This single-centre prospective pilot study included 30 couples between November 2018 and April 2019. PARTICIPANTS/MATERIALS, SETTING, METHOD: Arab couples, with a female age between 18-40 years, body mass index ≤30 kg/m2, at least 10 cumulus oocyte complexes (COCs) following oocyte retrieval (OR) and normal semen concentration and motility (WHO) in the fresh ejaculate on the day of OR, were eligible for the study. Half of the sibling oocytes were assigned to conventional IVF, and the other half were assigned to ICSI. All embryos were cultured in a time-lapse imaging system in Global Total LP media. Blastocysts were subjected to trophectoderm biopsy on Day 5, 6 or 7 and next-generation sequencing (NGS) to determine blastocyst ploidy status. The primary objective was to determine the euploid rate in blastocysts from sibling oocytes. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 568 COCs were randomly allocated between IVF (n = 283; 9.4 ± 4.0) and ICSI (n = 285; 9.5 ± 4.1). While the incidence of normal fertilisation per cycle (6.1 ± 3.8 (64.0%) vs 6.3 ± 3.5 (65.4%); P = 0.609) was distributed equally between IVF and ICSI, the degeneration rate (0.1 ± 0.3 vs 0.7 ± 0.8; P = 0.0003) was significantly higher after ICSI and the incidence of abnormal fertilisation (≥3 pronuclei) was significantly higher after IVF (0.9 ± 1.2 vs 0.2 ± 0.4; P = 0.005). For all fertilised oocytes, there were no differences in the number of good-quality embryos on Day 3 (74% vs 78%; P = 0.467), nor in the blastulation rate on Day 5 (80.4% vs 70.8%; P = 0.076). The total number of blastocysts biopsied per cycle on Days 5, 6 and 7 was not significantly different between IVF or ICSI (4.0 ± 2.8 vs 3.9 ± 2.5; P = 0.774). With euploid rates of 49.8 and 44.1% (P = 0.755; OR: 1.05664 [0.75188-1.48494), respectively, there was no significant difference identified between IVF and ICSI (2.0 ± 1.8 vs 1.9 ± 1.7; P = 0.808) and all couples had at least one euploid blastocyst available for transfer. When considering only euploid blastocysts, the male/female ratio was 61/39 in IVF and 43/57 in ICSI (P = 0.063). LIMITATIONS, REASON FOR CAUTION: This is a pilot study with a limited patient population of 30 couples (and 568 COCs) with a normal ovarian response. The results of our study should not be extrapolated to other patient populations. WIDER IMPLICATIONS OF THE FINDINGS: It is safe to apply conventional IVF in couples with non-male factor infertility undergoing PGT-A. STUDY FUNDING/COMPETING INTEREST(S): No funding was obtained. There are no competing interests. TRIAL REGISTRATION NUMBER: NCT03708991.

Vitamin D in Follicular Fluid Correlates With the Euploid Status of Blastocysts in a Vitamin D Deficient Population.

Context: The widespread distribution of the Vitamin D (VitD) receptor in reproductive tissues suggests an important role for VitD in human reproduction. The assessment of patient´s VitD is based on the 25-hydroxyvitamin D (25(OH)D) metabolite measurement. However, most of the circulating 25(OH)D is bound to either VitD-binding protein (VDBP) (88%) or albumin (12%) and less than 1% circulates free. Objective: To determine a possible correlation between VitD levels in serum (S) and follicular fluid (FF) and blastocyst ploidy status in patients undergoing infertility treatment. Methods: A prospective observational study was performed including couples planned for preimplantation genetic testing for aneuploidies (PGT-A) from ART Fertility Clinics. Patients were classified according to their 25(OH)D-Serum levels: VitD deficient group <20 ng/ml and insufficient/replete ≥20 ng/ml defined as VitD non-deficient group. Results: Serum samples and 226 FF from individual follicles were collected for 25(OH)D, bioavailable 25(OH)D, free 25(OH)D, and % free 25(OH)D measurement. 25(OH)D-Serum in VitD deficient and non-deficient were 13.2±4.0 ng/ml vs 32.3±9.2 ng/ml; p<0.001. FF from 40 and 74 biopsied blastocysts was analysed of which 52.5 and 60.8% were euploid (p = 0.428), respectively. In VitD deficient patients, mean 25(OH)D-FF, bioavailable 25(OH)D-FF, and free 25(OH)D-FF were higher in euploid vs aneuploid blastocysts (18.3±6.3 ng/ml vs 13.9±4.8 ng/ml; p = 0.040; 1.5±0.5 ng/ml vs 1.1±0.4 ng/ml; p = 0.015; 0.005±0.002 ng/ml vs 0.003±0.001 ng/ml; p = 0.023, respectively), whilst no differences were found in VitD non-deficient patients (37.9±12.3 ng/ml vs 40.6±13.7 ng/ml; p = 0.380; 3.1±1.1 ng/ml vs 3.3±1.2 ng/ml; p = 0.323; 0.01±0.003 ng/ml vs 0.01±0.004 ng/ml; p = 0.319, respectively). Conclusion: VitD non-deficient patients have a significantly higher probability of obtaining a euploid blastocyst compared to VitD deficient patients (OR:33.36, p = 0.002).

Using a Multi-Stage hESC Model to Characterize BDE-47 Toxicity During Neurogenesis.

Although the ramifications associated with polybrominated diphenyl ethers (PBDEs) exposures during human pregnancy have yet to be determined, increasing evidence in humans and animal models suggests that these compounds cause neurodevelopmental toxicity. Human embryonic stem cells (hESCs) models can be used to study the effects of environmental chemicals throughout the successive stages of neuronal development. Here, using a hESC differentiation model, we investigated the effects of common PBDE congeners (BDE-47 or -99) on the successive stages of early neuronal development. First, we determined the points of vulnerability to PBDEs across 4 stages of in vitro neural development by using assays to assess for cytotoxicity. Differentiated neural progenitors were identified to be more sensitive to PBDEs than their less differentiated counterparts. In follow-up investigations, we observed BDE-47 to inhibit functional processes critical for neurogenesis (eg, proliferation, expansion) in hESC-derived neural precursor cells (NPCs) at sub-lethal concentrations. Finally, to determine the mechanism(s) underlying PBDE-toxicity, we conducted global transcriptomic and methylomic analyses of BDE-47. We identified 589 genes to be differentially expressed due to BDE-47 exposure, including molecules involved in oxidative stress mediation, cell cycle, hormone signaling, steroid metabolism, and neurodevelopmental pathways. In parallel analyses, we identified a broad significant increase in CpG methylation. In summary our results suggest, on a cellular level, PBDEs induce human neurodevelopmental toxicity in a concentration-dependent manner and sensitivity to these compounds is dependent on the developmental stage of exposure. Proposed mRNA and methylomic perturbations may underlie toxicity in early embryonic neuronal populations.

Clinical reassessment of human embryo ploidy status between cleavage and blastocyst stage by Next Generation Sequencing.

One of the most important limitations of genetic testing in preimplantation embryos is embryonic mosaicism, especially when performed on D3 with only a single blastomere evaluated. Previous publications, using Array-Comparative Genomic Hybridization (a-CGH) to compare day 3 (D3) biopsies versus trophectoderm biopsies for the analysis of aneuploid embryos, showed similar high concordance rates per embryo diagnosis for D3 biopsies and trophectoderm biopsies. Next generation sequencing (NGS) was introduced lately as a new technique for preimplantation genetic testing for aneuploidies (PGT-A). Using this technique, this retrospective descriptive study evaluated the degree of the concordance of the diagnosis between preimplantation human cleavage stage (D3) and blastocyst stage (D5) embryos. Double biopsies on D3 and D5 were performed on 118 embryos, reaching blastocyst stage on D5 and had not been selected for transfer. As the fertilization law of the United Arab Emirates does not allow embryo freezing, also surplus euploid embryos after D 3 biopsy were included. Analysis of the NGS results from D3 and D5 embryo biopsies showed a total concordance rate per embryo diagnosis of 85.6% for euploid and aneuploid embryos. The concordance rates per embryo chromosomal pattern for embryo diagnosed as aneuploid at both biopsy stages was 82.2%. However, the status regarding the affected chromosomes was not identical on D3 and D5. Hence, the total concordance rate between D3 biopsy and D5 biopsy was limited to 67.8%. This current study clearly demonstrated that the concordance rates between D3 and D5 biopsies in aneuploid and euploid embryos are lower than previously reported.

Vitamin D deficiency and level of asthma control in women from North of Jordan: a case-control study.

INTRODUCTION: Reduced vitamin-D levels in patients with asthma have been associated with impaired lung function, increased airway hyper-responsiveness, and reduced glucocorticoid responsiveness. Nationwide studies revealed a considerable prevalence of vitamin-D deficiency (VDD) in Jordanian women. OBJECTIVE: A case-control study was conducted to determine the relationship between serum vitamin A and D levels and asthma among women in North of Jordan. METHODS: Sixty-eight asthmatics, age range between 14 and 65 years and 77 healthy women, age range between 19 and 51 years, were enrolled. Asthma severity was classified using Global Initiative for Asthma (GINA) guidelines and Asthma Control Test (ACT) questionnaire. Serum vitamin-A and 25-hydroxyvitamin-D (25(OH)D3) levels were measured using high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods, respectively. RESULTS: The prevalence of VDD (<15 ng/ml) was higher but not statistically significant for women with asthma compared with controls (95.6% vs. 87.0%; p = 0.070). The severity of VDD correlated with the number of asthma medications (p = 0.020). 25(OH)-D3 serum levels directly correlated with asthma control level using ACT score (p = 0.012) and GINA classification (p = 0.046). After adjusting for age, the odds of having VDD for asthmatic women were 35.9 times higher than that for women with no asthma. There was no difference in serum vitamin-A level between healthy and asthmatic women (p = 0.214) and none had vitamin-A deficiency (<200 µg/dl). CONCLUSIONS: VDD is prevalent in women with asthma in northern Jordan. The severity of VDD correlated with poor asthma control and a need for more medications to control asthma. There was no association between vitamin-A and asthma.

High prevalence rates of anemia, vitamin A deficiency and stunting imperil the health status of Bedouin schoolchildren in North Badia, Jordan.

AIMS: This survey aimed to identify nutritional deficits affecting Bedouin children as they enter school age and illustrate their families' dietary consumption patterns. METHODS: 560 healthy schoolchildren aged 5.5-10 years from 8 Badia hamlets were nutritionally investigated by applying anthropometric, laboratory and dietary assessment methods. RESULTS: Mean and median serum retinol concentrations (SRCs) were 228 and 218 microg/l, respectively. Mean prevalence of nutritional deficits among children were stunting (23.4%), anemia (57.5%), vitamin A deficiency (29.5%), iron deficiency (28.4%), and vitamin E deficiency (17.1%). Including those with borderline values, the proportion of children vulnerable to VAD threats reached 90%. Compared to normal subjects, anemic children had lower SRC (207 vs. 233 microg/l; p < 0.001) and ferritin (9.6 vs. 11.3 microg/l; p < 0.001) levels; stunted children had lower serum vitamin E (6.5 vs. 7.3 mg/l; p < 0.001) and ferritin (10.3 vs. 11.1 microg/l; p < 0.001) levels; vitamin A-deficient children had lower hemoglobin (11.3 vs. 11.8 g/dl; p < 0.001), serum vitamin E (6.69 vs. 7.23 mg/l; p < 0.01), and serum ferritin (10.4 vs. 11.2 microg/l; p < 0.001) levels, and vitamin E-deficient subjects had lower SRC levels (206 vs. 232 microg/l; p < 0.001). Except for hemoglobin status, no significant differences between genders were detected. SRC correlated strongly with hemoglobin, vitamin E, BMI and serum ferritin. The frequency of consumption of meat and fats seemed to be among the determinants of SRC and hemoglobin values. CONCLUSION: Urbanization has brought Bedouins towards poverty and undernutrition, with older children suffering more severe consequences in comparison with preschool-age children. This multifaceted nutritional problem requires implementation of multicomponent interventions.

Poor nutritional health of Bedouin preschool children in Jordan: the irony of urbanization.

AIMS: The purpose of this cross-sectional study was to assess growth and nutritional status of Bedouin preschool children with special focus on the extent of the interaction of anemia with iron deficiency and vitamin A deficiency as possible underlying factors. METHODS: Subjects aged 6-66 months were included by recruitment of 262 mothers having either an infant or a preschool child. Nutritional assessment by anthropometric, laboratory and dietary methods was carried out, with anthropometric indices expressed in z-score units. Serum retinol concentration, alpha-tocopherol, ferritin and blood hemoglobin and hematocrit were assessed. RESULTS: Stunting, vitamin A deficiency, anemia, and iron deficiency were prevalent at rates of 21.1, 25.2, 57.3, and 20.2%, respectively. Mean serum retinol concentration was 249 +/- 64 microg/l. More than 50% of the children were at risk of vitamin A deficiency. Compared to others, subjects with vitamin A deficiency tended to have lower hemoglobin (10.0 +/- 1.9 vs. 10.95 +/- 1.99 g/dl, p < 0.001) and lower serum ferritin concentrations (9.2 +/- 3.4 vs. 16.3 +/- 6.2 microg/l, p < 0.001). Serum retinol concentration correlated with hemoglobin (r = 0.41, p < 0.001) and ferritin (r = 0.42, p < 0.001). No gender differences were found. CONCLUSIONS: Bedouin preschool children are at risk for anemia, iron deficiency, nonocular vitamin A deficiency and stunting. Vitamin A deficiency exacerbates the development of anemia and iron deficiency among children. Urbanization has led Bedouins to the verge of poverty. They have been losing their characteristic lifestyle as traditional livestock keepers, with their children deprived of milk to drink or meat to eat. Coordinated interventional actions by the socioeconomic development planners and decision makers are needed.

High prevalence of subclinical vitamin A deficiency in Jordan: a forgotten risk.

This longitudinal study assessed growth and vitamin A status of schoolchildren after earlier surveys had linked stunting among Jordanian children to dietary zinc and iron inadequacies. A group of 1,023 subjects ages 5.5 to 9.9 years were randomly recruited for study from seven disadvantaged semirural districts. Baseline assessment included anthropometric and laboratory data with the relevant dietary information. Over nine months of study, the subjects received a daily snack meal. Immediately before the final assessment, each student received one 100,000 IU vitamin A capsule. At baseline there was a 19.9% prevalence of stunting, 18.8% for anemia, and 21.8% for subclinical vitamin A deficiency. Mean and median serum retinol concentrations were 248 (sd +/- 66) and 242 micrograms/L, respectively. In 98% of the cases, vitamin A-rich vegetables were consumed three or more days per week. About 60% of subjects had serum retinol levels in the range 200 to 300 micrograms/L. Only vitamin A foods from animal sources showed an influence (p < .05) on mean serum vitamin A values and growth score. Dietary and capsule supplementation had a significant positive impact only on serum retinol levels (p < .01) and on the anemia (p < .05) indicators. The conclusion underlines vitamin A deficiency among schoolchildren as a public health problem, and that the situation is anticipated to be more profound among preschool children, who are usually at greater risk of becoming deficient. Launching another, but controlled, intervention study in other sites, preferably with use of a tracer to rule out malabsorption in young children, is highly indicated.