Nevin Manimala

Planta. 2022 Jan 14;255(2):38. doi: 10.1007/s00425-022-03820-7.

ABSTRACT

Three known sugarcane aphid-resistant pollinator parents were sterilized in A3 cytoplasmic male sterility and were confirmed in this study to be resistant to sugarcane aphid allowing for the development of sugarcane aphid-resistant forage hybrids. We utilized A3 cytoplasmic male sterility and converted known sugarcane aphid-resistant sorghum TX 2783, and newly released R. LBK1 (Reg. No. GP-865, PI 687244) and R. LBK2 (Reg. No. GP-866, PI 687245) into A3 sterility to determine if the sterile counterparts would also equally express tolerance and or antibiosis to sugarcane aphid. Free-choice flat screen trials and life-table demographic studies were utilized and compared to know susceptible/fertile entries KS 585, and TX 7000, and known resistant/fertile entries TX 2783 and DKS 37-07. The R. LBK1 fertile entry was more tolerant than the known susceptible entries KS 585 and TX 7000, but was not as resistant as the other resistant entries, sustaining a damage rating of 6.0 across two different screen trials. The sterile A3 R. LBK2 showed a greater tolerance and expressed higher levels of antibiosis during aphid reproductive studies when compared to the known resistant and fertile TX 2783. All other fertile (R. LBK2, TX2783) and the A3 male sterile counterparts (A3 R. LBK2, A3 TX2783) were very similar in expression of high levels of tolerance and exhibited statistically similar damage ratings of 3.3-4.3 when exposed to sugarcane aphids. No entry, either fertile or sterile, was as tolerant as DKS 37-07, a known resistant commercial hybrid. Other plant measurements including percent loss in chlorophyll content, difference in plant height, and number of true leaves for sugarcane aphid infested versus non-infested were very consistent and highly correlated with damage ratings. Antibiosis was also exhibited in both fertile and sterile versions of the resistant lines. There was a 2 × reduction in fecundity between the R. LBK1 fertile and its sterile A3 R. LBK1 when compared to the susceptible KS 585 and TX 7000; however, the remaining fertile and sterile entries had 3.8 × to 5.8 × decrease in fecundity when compared to the susceptible KS 585 and TX 7000. Other measurements in life-table statistics such as nymphs produced/female/d, and the intrinsic rates of increased were significantly lower for all fertile and sterile lines, showing that antibiosis significantly affected sugarcane aphid reproduction. In conclusion, the A3 cytoplasmic male sterility shows consistency for maintaining the single dominant trait SCA-resistant trait of TX 2783 for expressing both antibiosis and tolerance, and great utility in the development of sugarcane aphid-resistant forage sorghums.

PMID:35031833 | DOI:10.1007/s00425-022-03820-7

Curr Protoc. 2022 Jan;2(1):e330. doi: 10.1002/cpz1.330.

ABSTRACT

Three-dimensional organoid cultures enable the study of stem cell and tissue biology ex vivo, providing improved access to cells for perturbation and live imaging. Typically, organoids are grown in hydrogel domes that are simple to prepare but that lead to non-uniform tissue growth and viability. We recently developed a simple alternative culture method to embed intestinal organoids in multilayered hydrogels, called “triple-decker sandwiches,” that align organoids in a common z-plane with uniform access to medium. This culture configuration improves the growth and survival of organoids over a wide working area and facilitates long-term confocal imaging and molecular perturbation. Here, we present protocols for preparing organoids in triple-decker sandwich cultures and using them for live imaging, immunostaining, and single-cell RNA sequencing. We have tested our methods on mouse and human intestinal organoids and expect them to be useful for other highly proliferative three-dimensional cell cultures. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Pre-coating plates with PolyHEMA to prepare them for triple-decker sandwich culture Support Protocol 1: Preparing PolyHEMA solution to coat glass-bottom dishes Basic Protocol 2: Embedding intestinal organoids in triple-decker sandwiches Alternate Protocol 1: Seeding single cells or organoids at low density in triple-decker sandwiches Support Protocol 2: Embedding intestinal organoids in hydrogel domes Support Protocol 3: Production of Wnt3a-conditioned medium Support Protocol 4: Production of Rspo1-conditioned medium Basic Protocol 3: Live imaging of mouse intestinal organoids in triple-decker sandwich cultures Alternate Protocol 2: Live imaging of vital dye-treated mouse intestinal organoids in triple-decker sandwich cultures Basic Protocol 4: Immunofluorescence imaging of mouse organoids liberated from triple-decker sandwich cultures Alternate Protocol 3: Liberating and fixing mouse intestinal organoids from dome cultures Support Protocol 5: Measuring cell proliferation by EdU staining of mouse intestinal organoids Basic Protocol 5: Single-cell RNA sequencing and analysis of mouse intestinal organoids.

PMID:35030297 | DOI:10.1002/cpz1.330

Int J Dent Hyg. 2022 Jan 14. doi: 10.1111/idh.12580. Online ahead of print.

ABSTRACT

INTRODUCTION: Parents and caregivers of pediatric patients usually tend to search for health information on social networks and other online platforms. At this point, the quality and reliability of these sources play an essential part in maintaining the oral health of pediatric patients.

AIM: The recent study aimed to analyze the reliability, quality, and content of YouTube^TM videos on pediatric oral health instructions, assess the efficacy of these videos, and help health providers lead the parents accessing accurate information on the subject mentioned.

DESIGN: The searching term was detected as “children oral health” (Google Trends Application). The first 150 videos were taken into the study. Six excluding criteria were used, and 40 videos have lasted for the further examinations. Content headings were determined according to the AAPD guidelines on pediatric oral health. The content analysis scores, reliability scores, and Global Quality Score were calculated for each of these 40 videos. SPSS 26 statistical software was used in the statistical analysis process of the data; p <0.05 was considered statistically significant.

RESULTS: The recent study results showed that according to content analyses, the videos were classified as ”poor” content with the mean values 4.07 (± 1.32). The mean reliability score 2.96 (± 0.69) has shown that the reliability of the videos was moderate. Mean Global Quality Score values were 2.95 (± 0.73), which means the subjects were moderate quality. The mean reliability of the rich content videos (3.39 ± 0.59) and the Global Quality Score score of them (3.43 ± 0.62) were statistically higher compared to the average values of the poor content videos (respectively; 2.73 ± 0.64; 2.69± 0.66) (p<0.05) The average number of likes, dislikes, length, interaction index and viewing rate of rich content videos were higher than the same features of poor content videos (p<0.05). The videos with rich content were determined in the first 50 videos listed for analyzes, and the distribution of the included videos on the list was equally.

CONCLUSION: The present study concluded that the YouTubeTM videos providing oral health instructions for pediatric patients and their parents had poor content, medium quality, and reliability. Although all the listed content headings were not mentioned in the videos, the content of them still may be useful and educational for individuals. However, dental hygiene practitioners and dentists should be aware of the need in this area and put more effort into improving the YouTube^TM videos on children’s oral health in a more detailed way regarding content headings, quality, and reliability.

PMID:35030292 | DOI:10.1111/idh.12580

Int Endod J. 2022 Jan 14. doi: 10.1111/iej.13681. Online ahead of print.

ABSTRACT

AIM: This preliminary randomised, prospective, controlled trial aimed to compare the clinical and radiographic outcomes of two regenerative endodontic procedures (REPs), revitalisation and a platelet-rich fibrin (PRF)-based technique, in the treatment of mature permanent teeth with necrotic pulps.

METHODOLOGY: The trial has been reported according to the Preferred Reporting Items for Randomised Trials in Endodontics 2020 guidelines. The study protocol was registered at the clinical trial registry (ClinicalTrials.gov) with identifier number NCT04158232. Twenty patients with mature necrotic anterior teeth with large periapical lesions were randomly allocated into two groups (n=10): group I, treated with revitalisation with the blood clot (BC) technique, and Group II, treated with a PRF-based technique. The follow-up was for 12 months. Periradicular healing was assessed using standardised radiographs taken at baseline, and at 6 and 12 months after treatment. An electric pulp tester was used to assess whether pulp sensibility had been regained during the follow-up period. Statistical analysis was conducted using Mann-Whitney test and Wilcoxon test for non-parametric data. For parametric data, repeated measures analysis of variance was used. The significance level was set at P≤0.05.

RESULTS: There was a significant increase in periradicular healing in both groups at 6 and 12 months, compared to that at baseline, with no significant difference between the studied groups after 12 months (P=0.143). There was a significant difference between the tooth sensibility readings at baseline, 6-month, and 12-month follow-up timepoints (P<0.001).

CONCLUSIONS: The findings of this preliminary trial indicate the potential for using REPs, such as revitalisation or PRF-based techniques, as treatment options for mature teeth with necrotic pulps. A higher level of evidence obtained through adequately powered clinical trials and longer follow-up periods are required to conclusively validate the different outcomes of REPs.

PMID:35030270 | DOI:10.1111/iej.13681

Am J Occup Ther. 2022 Jan 1;76(1):7601180100. doi: 10.5014/ajot.2022.049202.

ABSTRACT

IMPORTANCE: Rates of occupational therapy service utilization among people with autism spectrum disorder (ASD) or intellectual disability (ID) have not been explored in population-based samples.

OBJECTIVE: To describe occupational therapy services delivered to Medicaid-eligible persons younger than age 65 yr identified as having ASD, ID, or both and to evaluate demographic factors associated with occupational therapy service utilization in this population.

DESIGN: Retrospective, case-control, cohort study using claims records from Medicaid Analytic eXtract files (2009-2012).

SETTING: Data from all 50 states and Washington, DC.

PARTICIPANTS: Beneficiaries identified as having ASD only, ASD+ID, or ID only who were younger than age 18 yr (N = 664,214) and ages 18-64 yr (N = 702,338). Outcomes and Measures: We analyzed Current Procedural Terminology® and Healthcare Common Procedure Coding System procedure codes, Medicaid Statistical Information System type of service codes, and Center for Medicare & Medicaid Services provider specialty codes.

RESULTS: Only 3.7% to 6.3% of eligible adult beneficiaries received occupational therapy; in contrast, 20.5% to 24.2% of children received occupational therapy. Significant predictors of service use varied by group; however, differences by race-ethnicity, eligibility on the basis of poverty, and geographic location were observed. Among children, the most frequent billing code was for “therapeutic activities” (43%-60%); among adults, it was “community/work reintegration training” (29%-39%).

CONCLUSIONS AND RELEVANCE: Billed procedure code patterns do not consistently reflect the unique occupational focus that occupational therapy providers deliver to people with developmental disabilities. Disparities in occupational therapy receipt warrant further attention to understand the social and structural factors affecting service delivery. What This Article Adds: Occupational therapy services paid for by Medicaid are used more frequently by children with ASD and ID than by adults with these diagnoses. Greater understanding of the intersectional factors that drive service delivery and disparities is needed.

PMID:35030249 | DOI:10.5014/ajot.2022.049202

Clin Chem. 2022 Jan 14:hvab274. doi: 10.1093/clinchem/hvab274. Online ahead of print.

ABSTRACT

BACKGROUND: Droplet digital PCR (ddPCR) is a widely used and sensitive application for circulating tumor DNA (ctDNA) detection. As ctDNA is often found in low abundance, methods to separate low-signal readouts from noise are necessary. We aimed to characterize the ddPCR-generated noise and, informed by this, create a sensitive and specific ctDNA caller.

METHODS: We built 2 novel complimentary ctDNA calling methods: dynamic limit of blank and concentration and assay-specific tumor load estimator (CASTLE). Both methods are informed by empirically established assay-specific noise profiles. Here, we characterized noise for 70 mutation-detecting ddPCR assays by applying each assay to 95 nonmutated samples. Using these profiles, the performance of the 2 new methods was assessed in a total of 9447 negative/positive reference samples and in 1311 real-life plasma samples from colorectal cancer patients. Lastly, performances were compared to 7 literature-established calling methods.

RESULTS: For many assays, noise increased proportionally with the DNA input amount. Assays targeting transition base changes were more error-prone than transversion-targeting assays. Both our calling methods successfully accounted for the additional noise in transition assays and showed consistently high performance regardless of DNA input amount. Calling methods that were not noise-informed performed less well than noise-informed methods. CASTLE was the only calling method providing a statistical estimate of the noise-corrected mutation level and call certainty.

CONCLUSIONS: Accurate error modeling is necessary for sensitive and specific ctDNA detection by ddPCR. Accounting for DNA input amounts ensures specific detection regardless of the sample-specific DNA concentration. Our results demonstrate CASTLE as a powerful tool for ctDNA calling using ddPCR.

PMID:35030248 | DOI:10.1093/clinchem/hvab274

Am J Clin Nutr. 2022 Jan 14:nqac005. doi: 10.1093/ajcn/nqac005. Online ahead of print.

ABSTRACT

BACKGROUND: Visualizations of the emerging obesity epidemic, such as with serial United States (US) color prevalence maps, provide graphic images that extend informative public health messages beyond those in written communications. Advances in low-cost 3D optical technology now allows for development of large image databases that include participants varying in race/ethnicity, body mass, height, age, and circumferences. When combined with contemporary statistical methods, these data sets can be used to create humanoid avatar images with pre-specified anthropometric features.

OBJECTIVE: The current study aimed to develop a humanoid avatar series with characteristics of representative US adults extending over the past six decades.

METHODS: 3D optical scans were conducted on a demographically diverse sample of 570 healthy adults. Image data was converted to principal components and manifold regression equations were then developed with body mass, height, age, and waist circumference as covariates. Humanoid avatars were generated for representative adults with these four characteristics as reported in Center for Disease Control surveys beginning in 1960-62 up to 2015-18.

RESULTS: There was a curvilinear increase in adult US population body mass, waist circumference, and body mass index (BMI) in males and females across the nine surveys spanning six decades. A small increase in average adult population age was present between 1960 and 2018; height changes were inconsistent. A series of four avatars developed at approximately 20-year intervals for representative males and females reveal the changes in body size and shape consistent with the emergence of the obesity epidemic. An additional series of developed avatars portray the shapes and sizes of males and females at key BMI cut points.

CONCLUSIONS: New mathematical approaches and accessible 3D optical technology combined with increasingly available large and diverse data sets across the lifespan now makes unique visualization of body size and shape possible on a previously unattainable scale. The study is registered at: https://clinicaltrials.gov/ct2/show/NCT03637855.

PMID:35030235 | DOI:10.1093/ajcn/nqac005

Am J Clin Nutr. 2022 Jan 14:nqab422. doi: 10.1093/ajcn/nqab422. Online ahead of print.

ABSTRACT

BACKGROUND: Excessive vitamin A (VA) can cause bone resorption and impair growth. Government-mandated VA supplementation (VAS) and adequate intake through dietary fortification and liver consumption led to excessive VA in South African children.

OBJECTIVES: We evaluated the relationship of VAS and underlying hypervitaminosis A assessed by retinol isotope dilution (RID) with measures of growth and bone turnover in this cohort.

METHODS: Primary outcomes in these children (n = 94, 36-60 mo) were anthropometric measurements [height-for-age (HAZ), weight-for-age (WAZ), and weight-for-height (WHZ) Z-scores]; serum bone turnover markers [C-terminal telopeptide of type I collagen (CTX) and N-terminal propeptide of type I procollagen (P1NP)]; and inflammation defined as C-reactive protein (CRP ≥ 5 mg/L) and/or α1-acid glycoprotein (AGP ≥1 g/L). VA status was previously measured by RID-estimated total body VA stores (TBSs) and total liver VA reserves (TLRs), and serum retinol and carotenoid concentrations, before and 4 wk after children were administered 200,000 IU VAS. Serum 25-hydroxyvitamin D3 was measured by ultra-performance LC. (Registered at Clinicaltrials.gov NCT02915731).

RESULTS: In this largely hypervitaminotic A cohort, HAZ, WAZ, and WHZ were negatively associated with increasing TLRs, where TLRs predicted 6-10% of the variation before VAS (P < 0.05), increasing to 14-19% 4 wk after VAS (P < 0.01). Bone resorption decreased after VAS (P < 0.0001) while formation was unaffected. Neither CTX nor P1NP were correlated with TLRs at either time. Serum carotenoids were low. One child at each timepoint was vitamin D deficient (<50 nmol/L). CRP and AGP were not associated with growth measurements.

CONCLUSIONS: Excessive TLRs due to dietary vitamin A intake and supplementation is associated with lower anthropometric measures and bone resorption decreased after supplementation. Vitamin A supplementation programs should monitor VA status with biomarkers sensitive to TLRs to avoid causing negative consequences in children with hypervitaminosis A.The Clinical Trial Registry number is NCT02915731: https://clinicaltrials.gov/ct2/show/NCT02915731.

PMID:35030234 | DOI:10.1093/ajcn/nqab422

PLoS One. 2022 Jan 14;17(1):e0262584. doi: 10.1371/journal.pone.0262584. eCollection 2022.

ABSTRACT

The equine neonate is considered to have impaired glucose tolerance due to delayed maturation of the pancreatic endocrine system. Few studies have investigated insulin sensitivity in newborn foals using dynamic testing methods. The objective of this study was to assess insulin sensitivity by comparing the insulin-modified frequently sampled intravenous glucose tolerance test (I-FSIGTT) between neonatal foals and adult horses. This study was performed on healthy neonatal foals (n = 12), 24 to 60 hours of age, and horses (n = 8), 3 to 14 years of age using dextrose (300 mg/kg IV) and insulin (0.02 IU/kg IV). Insulin sensitivity (SI), acute insulin response to glucose (AIRg), glucose effectiveness (Sg), and disposition index (DI) were calculated using minimal model analysis. Proxy measurements were calculated using fasting insulin and glucose concentrations. Nonparametric statistical methods were used for analysis and reported as median and interquartile range (IQR). SI was significantly higher in foals (18.3 L·min-1· μIU-1 [13.4-28.4]) compared to horses (0.9 L·min-1· μIU-1 [0.5-1.1]); (p < 0.0001). DI was higher in foals (12 × 103 [8 × 103-14 × 103]) compared to horses (4 × 102 [2 × 102-7 × 102]); (p < 0.0001). AIRg and Sg were not different between foals and horses. The modified insulin to glucose ratio (MIRG) was lower in foals (1.72 μIUinsulin2/10·L·mgglucose [1.43-2.68]) compared to horses (3.91 μIU insulin2/10·L·mgglucose [2.57-7.89]); (p = 0.009). The homeostasis model assessment of beta cell function (HOMA-BC%) was higher in horses (78.4% [43-116]) compared to foals (23.2% [17.8-42.2]); (p = 0.0096). Our results suggest that healthy neonatal foals are insulin sensitive in the first days of life, which contradicts current literature regarding the equine neonate. Newborn foals may be more insulin sensitive immediately after birth as an evolutionary adaptation to conserve energy during the transition to extrauterine life.

PMID:35030228 | DOI:10.1371/journal.pone.0262584

PLoS One. 2022 Jan 14;17(1):e0262425. doi: 10.1371/journal.pone.0262425. eCollection 2022.

ABSTRACT

In this study, we examined the total bacterial community associated with ureolytic biomineralization from urine drainage systems. Biomineral samples were obtained from 11 California Department of Transportation public restrooms fitted with waterless, low-flow, or conventional urinals in 2019. Following high throughput 16S rRNA Illumina sequences processed using the DADA2 pipeline, the microbial diversity assessment of 169 biomineral and urine samples resulted in 3,869 reference sequences aggregated as 598 operational taxonomic units (OTUs). Using PERMANOVA testing, we found strong, significant differences between biomineral samples grouped by intrasystem sampling location and urinal type. Biomineral microbial community profiles and alpha diversities differed significantly when controlling for sampling season. Observational statistics revealed that biomineral samples obtained from waterless urinals contained the largest ureC/16S gene copy ratios and were the least diverse urinal type in terms of Shannon indices. Waterless urinal biomineral samples were largely dominated by the Bacilli class (86.1%) compared to low-flow (41.3%) and conventional samples (20.5%), and had the fewest genera that account for less than 2.5% relative abundance per OTU. Our findings are useful for future microbial ecology studies of urine source-separation technologies, as we have established a comparative basis using a large sample size and study area.

PMID:35030221 | DOI:10.1371/journal.pone.0262425