Adjuvant measures and genetic evaluations to improve results of hair transplantation

Follicular unit extraction (FUE) has evolved dramatically as the most recent advancement in surgical hair restoration as it leaves a tiny scar and creates natural and pleasing results. This study aims to show the effectiveness of adjuvant measures and genetic evaluations in improving outcomes. Prospective analysis of 271 male patients with androgenic alopecia who underwent hair transplantation with FUE technique between August 2015 and February 2020 at our center was conducted. The mean age was 35.93 ±4.40 years. At one year postoperatively, patients were asked to fill up a questionnaire which included their satisfaction level, need for 2nd session, and complications. Informed written consent was obtained from all patients. Also, blood samples were provided from patients before the operation. RNA extraction and cDNA synthesis were performed using the RNX-Plus kit (Cinnagen, Iran) and Vivantis kit (Malaysia). Amplification of SRD5A2 and GAPDH genes (as internal standard) for measuring gene expression was performed by real-time PCR. Data were analyzed using the statistical package for social science SPSS V. 23. In the last 156 cases, the addition of 40 mg of Triamcinolone to the LA solution led to a dramatic reduction of the incidence of postoperative oedema, from 40% to 9%. Adding three sessions of PRP at 2nd, 4th and 6th months postoperatively resulted in an increased patient satisfaction rate with better hair density and thickness where the rate of highly satisfied patients increased from 64.5% to 83.7%. The addition of 40 mg Triamcinolone to the LA solution was highly effective in reducing postoperative oedema. Three sessions of PRP at 2nd, 4th and 6th months postoperatively were recommended. The results of SRD5A2 gene expression showed that the expression of this gene in satisfied (P = 0.049) and dissatisfied (P = 0.028) patients were significantly higher than highly satisfied patients, which means that the SRD5A2 gene expression had an essential role in the successfulness of hair transplantation. The increased expression of this gene could reduce the response to hair transplantation.

Childhood Obesity and Risk of Stroke: A Mendelian Randomisation Analysis

Background: The causal relationship between childhood obesity and stroke remains unclear. Our objective was to elucidate the causal relationship between childhood obesity and the risk of stroke and its subtypes by performing Mendelian randomisation (MR) analyses.Methods: Genetic instruments for childhood obesity were obtained from a genome-wide association study (GWAS) of 13,848 European participants. Summary level data for stroke, intracerebral haemorrhage, ischaemic stroke (IS), and its subtypes were evaluated using the MEGASTROKE GWAS dataset, which included 446,696 European adults. Inverse-variance weighting, weighted-median analysis, MR-Egger regression, MR Pleiotropy RESidual Sum and Outlier test (MR-PRESSO), and MR-Robust Adjusted Profile Score were applied in this MR analysis. The leave-one-out sensitivity test, MR-PRESSO Global test, and Cochran’s Q test were conducted to confirm the accuracy and robustness of our results.Results: Genetic evaluations revealed that childhood obesity was associated with a higher risk of stroke (OR = 1.04, 95%CI: 1.01–1.07, p = 0.005) and IS (OR = 1.05, 95%CI: 1.02–1.08, p = 0.003), but not with intracerebral haemorrhage (ICH, OR = 0.93, 95%CI: 0.80–1.09, p = 0.39). In the subtype analysis, childhood obesity was also associated with large artery stroke (LAS, OR = 1.12, 95%CI: 1.02–1.22, p = 0.016) but not with cardioembolic stroke (OR = 1.06, 95%CI: 0.96–1.18, p = 0.21) and small vessel stroke (OR = 1.06, 95%CI: 0.98–1.15, p = 0.17). These results were stable in the sensitivity analysis and remained significant after Bonferroni correction.Conclusion: Our study provides evidence that childhood obesity is associated with a higher risk of stroke, IS, and LAS. The prevention of stroke, especially IS and LAS, should be promoted in populations with childhood obesity.

Cytotoxic, apoptotic, and genetic evaluations of Nigella sativa essential oil nanoemulsion against human hepatocellular carcinoma cell lines

Background Phytochemicals and plant extracts are showing promising anticancer potentials. In the current study, the volatile faction (essential oil) of Nigella sativa seeds was evaluated against some hepatocellular carcinoma (HCC). The essential oil was extracted and characterized by chromatographic techniques to reveal its chemical composition, especially thymoquinone. Then, the oil was fabricated in two nanoemulsion formulations (F1 and F2), which differ in their composition of surfactants. The cytotoxicity and apoptotic activities of the essential oil and its nanoemulsions were evaluated in vitro against HepG2 and Huh-7 cell lines. Normal WI-38 cell line was also included in that evaluation to study the selectivity and safety of the different formulations on normal cells. Results Gas chromatographic analysis indicated that the essential oil is composed mainly of p-cymene (40.0%), thymoquinone (31.2%) and trans-α-thujene (12.8%). Particle size of the nanoemulsions ranged between 9.4 and 119.7 nm depending on the type of surfactant used in the formulation process. The pure essential oil and its two nanoemulsions (F1 and F2) showed dose-dependent antiproliferative activity against both HCC cells. This activity reached its highest cell inhibition in the case of nanoemulsion (F2) where the proliferation percentage was only 21.9% and 9.2% against HepG2 and Huh-7 cells, respectively. The same nanoemulsion (F2) also showed the lowest IC50 values (55.7 and 35.5 µg/ml) against both HepG2 and Huh-7 cells, respectively, compared to 100 µg/ml for the reference drug Doxorubicin. Flow cytometric analysis also confirmed that nanoemulsion (F2) has the highest apoptotic activity compared to nanoemulsion (F1) and the pure unformulated essential oil. Genetic expressions of pro-apoptotic (Bax) and the anti-apoptotic (Bcl-2) gene markers evaluation revealed that nanoemulsion (F2) has better activity in upregulating (Bax) and down-regulate (Bcl-2) with the highest Bax/Bcl-2 ratio (69) was found against Huh-7 cells. All N. sativa nanoemulsions showed minimal cytotoxicity on the normal WI-38 cell, indicating wide safety margins due to selective properties. Conclusion Overall, the study revealed the potentials of N. sativa essential oil, after formulation in specially tailored nanoemulsion for application as potential adjuvant liver anticancer agent. Graphical Abstract

Genotypic Variation and Talent Identification in Sports

Top performance of athletes is not limited to the demand of fame, public recognition, sponsorship, and prize money but genetic inheritance contributes a prime role to hold such traits. Recent years, we have witnessed the rise of sports specific tests that identify person’s athletic talents, but human vary on genetic factors which silently work to achieve success in sports. Recent progress on the genetic determination in the sports sciences offer great perspective to analyze the genotype profile associated with the athletes. One of the most used advances in this field is the identification of variations in the DNA sequence, known as Single Nucleotide Polymorphisms (SNPs). Genetic evaluations should be combined with other tools to get an accurate identification of athletes and their respective fields to achieve optimum success.

Bias in variance component estimation in swine crossbreeding schemes using selective genotyping and phenotyping strategies

Selective genotyping of crossbred (CB) animals to include in traditionally purebred (PB) dominated genetic evaluations has been shown to provide an increase in the response to selection for CB performance. However, the inclusion of phenotypes from selectively genotyped CB animals, without the phenotypes of their non-genotyped cohorts, could cause bias in estimated variance components (VC) and subsequent estimated breeding values (EBV). The objective of the study was to determine the impact of selective CB genotyping on VC estimates and subsequent bias in EBV when non-genotyped CB animals are not included in genetic evaluations. A swine crossbreeding scheme producing 3-way CB animals was simulated to create selectively genotyped datasets. The breeding scheme consisted of three PB breeds each with 25 males and 450 females, F1 crosses with 1200 females and 12,000 CB progeny. Eighteen chromosomes each with 100 QTL and 4k SNP markers were simulated. Both PB and CB performance were considered to be moderately heritable (h2=0.4). Factors evaluated were, 1) CB phenotype and genotype inclusion of 15% (n=1800) or 35% (n=4200), 2) genetic correlation between PB and CB performance (rpc=0.1, 0.5 or 0.7) and 3) selective genotyping strategy. Genotyping strategies included: a) Random: random CB selection, b) Top: highest CB phenotype and c) Extreme: half highest and half lowest CB phenotypes. Top and Extreme selective genotyping strategies were considered by selecting animals in full-sib (FS) families or among the CB population (T). In each generation, 4320 PB selection candidates contributed phenotypic and genotypic records. Each scenario was replicated 15 times. VC were estimated for PB and CB performance utilizing bivariate models using pedigree relationships with dams of CB animals considered to be unknown. Estimated values of VC for PB performance were not statistically different from true values. Top selective genotyping strategies produced deflated estimates of phenotypic VC for CB performance compared to true values. When using estimated VC, Top_T and Extreme_T produced the most biased EBV, yet EBV of PB selection candidates for CB performance were most accurate when using Extreme_T. Results suggest that randomly selecting CB animals to genotype or selectively genotyping Top or Extreme CB animals within full-sib families can lead to accurate estimates of additive genetic VC for CB performance and unbiased EBV.

PSXV-1 Genetic evaluation of longevity of cows culled due to fertility-related problems using random regression models and censored data

A recent study showed that longevity based on different culling reasons should be considered as different traits in genetic evaluations. However, it is still necessary to create a pipeline that avoid including/excluding animals culled for different reasons in every genetic evaluation run. This study aimed to: 1) perform a genetic evaluation of the longevity of cows culled due to fertility-related problems including records of animals culled for other reasons (i.e., age, structural problems, disease, and performance) as censored records; and, 2) identify the impact of censored data in the genetic parameters and breeding values estimated. Two longevity indicators were evaluated: traditional (TL; time from first calving to culling) and functional (FL; time period in which the cow was alive and also calving after its first calving) longevity. Both TL and FL were evaluated from 2 to 15 years-old, and codified as binary traits for each age (0 = culled and 1 = alive/calved). Both trait definitions were analyzed using a Bayesian random regression linear model. Animals culled for reasons other than fertility were either excluded from the data (standard) or had their records censored after the culling date reported in the dataset (censored). After the quality control, 154,419 and 450,124 animals had uncensored and censored records, respectively. Heritabilities estimated for TL over the ages ranged from 0.02 to 0.13 for standard, and from 0.01 to 0.12 for censored datasets. Heritabilities estimated for FL ranged from 0.01 to 0.14 (standard), and from 0.01 to 0.13 (censored). Average (SD) correlation of breeding values predicted over all ages, using the standard and censored datasets, was 0.77 (0.16) for TL, and 0.83 (0.11) for FL. Our findings suggest that including censored data in the analyses might impact the genomic evaluations and further work is need to determine the optimal predictive approach.

33 Impact of Including the Cause of Missing Records on Genetic Evaluations for Growth in Commercial Pigs

It is of interest to evaluate crossbred pigs for hot carcass weight (HCW) and birth weight (BW); however, obtaining a HCW record is dependent on livability (LIV) and retained tag (RT). The purpose of this study is to analyze how HCW evaluations are affected when herd removal and missing identification are included in the model and examine if accounting for the reasons for missing traits improves the accuracy of predicting breeding values. Pedigree information was available for 1,965,077 purebred and crossbred animals. Records for 503,716 commercial three-way crossbred terminal animals from 2014 to 2019 were provided by Smithfield Premium Genetics. Two pedigree-based models were compared; model 1 (M1) was a threshold-linear model with all four traits (BW, HCW, RT, and LIV), and model 2 (M2) was a linear model including only BW and HCW. The fixed effects used in the model were contemporary group, sex, age at harvest (for HCW only), and dam parity. The random effects included direct additive genetic and random litter effects. Accuracy, dispersion, bias, and Pearson correlations were estimated using the linear regression method. The heritabilities were 0.11, 0.07, 0.02, and 0.04 for BW, HCW, RT, and LIV, respectively, with standard errors less than 0.01. No difference was observed in heritabilities or accuracies for BW and HCW between M1 and M2. Accuracies were 0.33, 0.37, 0.19, and 0.23 for BW, HCW, RT, and LIV respectively. The genetic correlation between BW and RT was 0.34 ± 0.03, and between BW and LIV was 0.56 ± 0.03. The positive and moderate genetic correlations between BW and other traits imply a heavier BW resulted in a higher probability of surviving to harvest. Despite the heritable and correlated aspects of RT and LIV, results imply no major differences between M1 and M2; hence, it is unnecessary to include these traits in classical models for BW and HCW.

PSVIII-7 Genetic parameters for health traits in dairy calves

Recent issues in the dairy industry related to both animal and public health concerns are leading farmers away from the use of drugs, while placing more focus on animal health and welfare. Public demands are also shifting towards ensuring socially acceptable production practices in terms of good animal health and welfare. Such challenges are moving the focus in dairying from solely financial to a broader set of themes that, once addressed, will enhance the sustainability of dairying and provide a long-term competitive advantage for the Canadian industry. Furthermore, from a production standpoint, calf diseases, such as, diarrhea and respiratory disease (RD) have been associated with decreased first lactation production and growth rate, therefore decreasing an animal’s potential lifetime profitability. As part of a larger project aiming to add calf health traits to genetic evaluations in Canadian dairy cattle, this study provides the groundwork through the estimation of genetic parameters of two calf health traits, diarrhea and RD. Data were provided by Lactanet Canada, and included 20,594 calf records for diarrhea from 741 herds, and 48,927 calf records for RD from 1,412 herds, recorded between 2004 and 2021 across Canada. Total herd records ranged between 1 and 3,860 for RD with an average of 37 records per herd, while for diarrhea records ranged between 1 and 3,724 with an average of 28 records per herd. The results of this study will be used to optimally fit both diarrhea resistance and RD resistance into a novel resiliency index for use in national genetic evaluations in Canada.

26 Are Subjectively Scored Linear Type Traits Suitable Predictors of Genetic Merit for Feed Intake in Grazing Holstein-friesian Dairy Cows?

Measuring dry matter intake (DMI) in grazing dairy cows using currently available techniques is invasive, time consuming, and expensive. An alternative to directly measuring DMI for use in genetic evaluations is to identify a set of readily available data sources that can be used in a multi-trait genetic evaluation with DMI. The objectives of the present study were to estimate the genetic correlations between readily available body-related linear type trait information and DMI in grazing, lactating Irish dairy cows and to estimate the partial genetic correlations between linear traits and DMI, after adjusting for differences in genetic merit for body weight. After edits, a total of 8,055 test-day records of DMI, body weight, and milk yield from 1,331 multiparous dairy cows were available, as were chest width, body depth, and stature scores for 47,141 first lactation dairy cows. In addition to considering the routinely recorded linear type traits individually, novel traits were defined as the product of two or three linear type traits as an approximation of rumen volume. The genetic variance of DMI, body weight, milk yield, and linear type traits were estimated using univariate animal linear mixed models. Sire linear mixed models were used to calculate genetic and phenotypic covariances. All linear type traits were moderately heritable (0.27 to 0.49) and genetically correlated (0.29 to 0.63) with DMI. The genetic correlations between the individual linear type traits and DMI, when the latter was adjusted for differences in the genetic merit for body weight, varied from 0.00 to 0.39. If the (partial) genetic correlations were validated with genetic evaluations, routinely available linear type trait records and live weight data could facilitate the selection of DMI in dairy cows, removing the need to capture large amounts of cost prohibitive feed intake phenotypes.

Cytotoxic, Apoptotic, and Genetic Evaluations of Nigella Sativa Essential Oil Nanoemulsion Against Human Hepatocellular Carcinoma Cell Lines

Background: Phytochemicals and plant extracts are showing promising anticancer potentials. In the current study the volatile faction (essential oil) of Nigella sativa seeds was evaluated against some hepatocellular carcinoma (HCC). The essential oil was extracted and characterized by chromatographic techniques to reveal its chemical composition, especially thymoquinone. Then, the oil was fabricated in two nanoemulsion formulations (F1 and F2), which differ in their composition of surfactants. The cytotoxicity and apoptotic activities of the essential oil and its nanoemulsions were evaluated in vitro against HepG2 and Huh-7 cell lines. Normal WI-38 cell line was also included in that evaluation to study the selectivity and safety of the different formulations on normal cells. Results: Gas chromatographic analysis indicated that the essential oil is composed mainly of p-cymene (40.0%), thymoquinone (31.2%) and trans-α-thujene (12.8%). Particle size of the nanoemulsions ranged between 9.4 nm to 119.7 nm depending on the type of surfactant used in the formulation process. The pure essential oil and its two nanoemulsions (F1 & F2) showed does-dependent antiproliferative activity against both HCC cells. This activity reached its highest cell inhibition in the case of nanoemulsion (F2) where the proliferation percentage was only 21.9% and 9.2% against HepG2 and Huh-7 cells, respectively. The same nanoemulsion (F2) also showed the lowest IC50 values (55.7 and 35.5 µg/ml) against both HepG2 and Huh-7 cells, respectively, compared to 100 µg/ml for the reference drug Doxorubicin. Flow cytometric analysis also confirmed that nanoemulsion (F2) has the highest apoptotic activity compared to nanoemulsion (F1) and the pure unformulated essential oil. Genetic expressions of pro-apoptotic (Bax) and the anti-apoptotic (Bcl-2) gene markers evaluation revealed that nanoemulsion (F2) has better activity in up-regulating (Bax) and down-regulate (Bcl-2) with the highest Bax/Bcl-2 ratio (69) was found against Huh-7 cells. All N. sativa nanoemulsions showed minimal cytotoxicity on the normal WI-38 cell, indicating wide safety margins due to selective properties. Conclusion: Overall, the study revealed the potentials of N. sativa essential oil, after formulation in specially tailored nanoemulsion for application as potential adjuvant liver anticancer agent.