Genome-wide association analysis of eggshell color of an F2 generation population reveals candidate genes in chickens.

Eggshell color is an important visual characteristic that affects consumer preferences for eggs. Eggshell color, which has moderate to high heritability, can be effectively enhanced through molecular marker selection. Various studies have been conducted on eggshell color at specific time points. However, few longitudinal data are available on eggshell color. Therefore, the objective of this study was to investigate eggshell color using the Commission International de L'Eclairage L*a*b* system with multiple measurements at different ages (age at the first egg and at 32, 36, 40, 44, 48, 52, 56, 60, 66, and 72 weeks) within the same individuals from an F2 resource population produced by crossing White Leghorn and Dongxiang Blue chicken. Using an Affymetrix 600 single nucleotide polymorphism (SNP) array, we estimated the genetic parameters of the eggshell color trait, performed genome-wide association studies (GWASs), and screened for the potential candidate genes. The results showed that pink-shelled eggs displayed a significant negative correlation between L* values and both a* and b* values. Genetic heritability based on SNPs showed that the heritability of L*, a*, and b* values ranged from 0.32 to 0.82 for pink-shelled eggs, indicating a moderate to high level of genetic control. The genetic correlations at each time point were mostly above 0.5. The major-effect regions affecting the pink eggshell color were identified in the 10.3-13.0 Mb interval on Gallus gallus chromosome 20, and candidate genes were selected, including SLC35C2, PCIF1, and SLC12A5. Minor effect polygenic regions were identified on chromosomes 1, 6, 9, 12, and 15, revealing 11 candidate genes, including MTMR3 and SLC35E4. Members of the solute carrier family play an important role in influencing eggshell color. Overall, our findings provide valuable insights into the phenotypic and genetic aspects underlying the variation in eggshell color. Using GWAS analysis, we identified multiple quantitative trait loci (QTLs) for pink eggshell color, including a major QTL on chromosome 20. Genetic variants associated with eggshell color may be used in genomic breeding programs.

Effects of manganese glycine on eggshell quality, eggshell ultrastructure, and elemental deposition in aged laying hens.

Poor eggshell quality of eggs laid by aged laying hens is the major problem affecting the length of the rearing period in the laying hen industry. Trace elements are required and play vital roles in the eggshell quality of laying hens. Appropriate dose of organic microelements is environmentally friendly and sufficient to satisfy the needs of hens because of their greater bioavailability and lower excretion than inorganic forms. The aim of this experiment was to investigate the effects of manganese (Mn) glycine (MG) on eggshell quality, elemental deposition, and eggshell ultrastructure in aged laying hens. A total of 720 Hy-Line Brown hens 70 weeks old were assigned equally to four groups with six replicates of 30 birds each. The hens were fed basal diets (without Mn supplementation) supplemented with 120 mg/kg of Mn from manganese sulfate monohydrate (MSM), or 40, 80, or 120 mg/kg Mn from MG for 12 weeks. Dietary supplementation with 80 mg/kg Mn from MG resulted in the greatest eggshell strength after 6 weeks of treatment (P = 0.047), and in greater eggshell strength than observed in the MSM control after 12 weeks of treatment (P = 0.025). After 12 weeks of treatment, the eggs of hens in the MG groups showed lower mammillary layer thickness in the blunt end, equator, and acute end than observed in the MSM control group (P < 0.001). With the exception of the blunt ends of eggs from hens in the 120 mg/kg MG group, the eggs of hens in the MG groups, compared with the MSM control group, exhibited a lower mammillary layer ratio, and greater palisade layer ratio and effective layer ratio in the blunt end, equator, and acute end (P < 0.001). Dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control and 40 and 120 mg/kg MG, resulted in the greatest palisade layer thickness and effective layer thickness, and the lowest mammillary layer thickness in the equator (P < 0.001, P = 0.001, P < 0.001, respectively). Furthermore, supplementation with 80 mg/kg Mn from MG exhibited the greatest ratio of the palisade layer and effective layer, and the lowest mammillary layer ratio in the blunt end and equator (all P < 0.001). The Mn content of eggshells in hens-fed diets supplemented with 80 and 120 mg/kg Mn from MG was greater than that in the MSM control and 40 mg/kg MG groups (P = 0.035). Dietary supplementation with 80 or 120 mg/kg Mn from MG resulted in greater tibia Mn content than observed in the 40 mg/kg MG group (P = 0.019), and greater yolk Mn content than observed in the 40 mg/kg MG and MSM control groups (P = 0.018). In conclusion, dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control (120 mg/kg Mn), may increase the deposition efficiency of Mn, alter eggshell elemental composition, improve eggshell ultrastructure, and enhance eggshell strength in aged laying hens.

Effects of exogenous energy on synthesis of steroid hormones and expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen.

Energy and the cAMP-response element binding protein (CREB)/steroidogenic acute regulatory protein (StAR) signaling pathway play important roles in steroid hormone production and follicular development in hens. This present study aimed to investigate the effects of exogenous energy on the synthesis of steroid hormones and the expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen. The primary theca cells of small yellow follicles were randomly divided into 6 treatments and cultured in medium with glucose concentrations of 1, 1.5, 3, 4.5, 6, and 7.5 mg/mL for 48 h. It was found that growth was robust and cell outlines were clear when cells were cultured with 1, 1.5, 3, and 4.5 mg/mL glucose, but cell viability was diminished and cell density decreased after exposure to glucose at 6 and 7.5 mg/mL for 48 h. Cell viability showed an increasing and then decreasing quadratic response to increasing glucose concentration in culture (r2 = 0.688, P < 0.001). The cell viability of theca cells cultured with 4.5 mg/mL glucose was greater than those cultured with 1, 1.5, 6, and 7.5 mg/mL glucose (P < 0.05). The concentration of estradiol in the medium containing 3 mg/mL glucose was higher than in medium containing 1, 1.5, and 6 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between progesterone concentrations and glucose concentrations (r2 = 0.522, P = 0.002). The concentration of progesterone in medium with 4.5 mg/mL glucose was higher than in medium with 1 and 7.5 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between the relative expression of CREB1 (r2 = 0.752, P < 0.001), StAR (r2 = 0.456, P = 0.002), CYP1B1 (r2 = 0.568, P < 0.001), and 3ß-HSD (r2 = 0.319, P = 0.018) in theca cells of laying hens and glucose concentrations after treatment with different glucose concentrations for 48 h. After treatment with 4.5 mg/mL glucose, the expression of StAR, CYP1B1, and 3ß-HSD genes were increased compared to treatment with 1, 1.5, 3, 6, and 7.5 mg/mL glucose (P < 0.001). There was an increasing and then decreasing quadratic correlation between glucose concentrations and protein expression of CREB1 (r2 = 0.819, P < 0.001), StAR (r2 = 0.844, P < 0.001), 3ß-HSD (r2 = 0.801, P < 0.001), and CYP11A1 (r2 = 0.800, P < 0.001) in theca cells of laying hens. The protein expression of CREB1, StAR, and 3ß-HSD in theca cells cultured with 4.5 mg/mL glucose was higher than in other groups (P < 0.001). The results indicate that the appropriate glucose concentration (4.5 mg/mL) can improve the synthesis of steroid hormones in theca cells of laying hens through the upregulation of key genes and proteins in the CREB/StAR signaling pathway.

Effects of energy restriction during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk.

The aim of this study was to assess the effects of energy-restricted feeding during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk. A total of 720 six-week-old layer chicks were allocated equally to 3 groups with 6 replicates of 40 pullets each, and were fed 1 of 3 diets that were nutritionally similar except for the apparent metabolizable energy corrected for nitrogen (AMEn) content. At the age of 6 to 17 wk, the pullets in the control group were given diet with 2,850 kcal/kg AMEn, and were fed ad libitum. The levels of AMEn in diet of pullets in the experimental groups were 90% (2,565 [2,850 × 90%] kcal/kg) and 80% (2,280 [2,850 × 80%] kcal/kg) of that in control group, and the daily amount of feed was restricted to the absolute quantity of the diet consumed by pullets in control group. At the age of 18 to 72 wk, all the hens were fed with the same diets ad libitum. As energy restriction increased in the growing phase, body weight (BW) dropped at the ages of 12 and 15 to 23 wk (at 23 wk: P = 0.001; at other ages: P < 0.001), but it showed no significant difference at 24 wk (P = 0.071). At 20 wk, restricting energy induced a delay in the development of sexual organs, including the ovary stroma, oviduct, and small yellow follicle (P < 0.05), as well as a delay in sexual maturity (P < 0.05). Consequently, the laying rate in the first and second periods dropped linearly (P = 0.046, 0.030, and 0.038, P < 0.001, respectively). The coefficient of variation (CV) in the BW at 19, 20, and 21 wk (P = 0.040, 0.023, and 0.042, respectively), the CV of age at first egg (P < 0.001), and CV of individual egg number at age 18 to 72 wk (P < 0.001) decreased linearly. There was a linear increase in the laying rate of hens in the later periods (at age 32-72 wk, P < 0.05), as well as in the average total egg number per hen and average laying rate at the age of 18 to 72 wk (P = 0.006). The average egg mass also showed a linear increase with increasing levels of energy restriction (P < 0.001). In summary, although appropriate energy restriction during growing phase delayed sexual maturity and sexual organ development in early-laying Hyline Brown pullets, it improved uniformity of BW, age at first egg laying, and individual egg number, and increased egg number per hen, laying rate, average egg mass, and number of settable eggs from 18 to 72 wk of age.

[Concordant systemic and local eosinophilia relates to poorer disease control in patients with nasal polyps].

Objective: To explore whether blood and polyp tissue eosinophil numbers are independent risk factors for poor disease control in patients with nasal polyp. Methods: By using the electronic medical records database and manual evaluation, 183 nasal polyp patients who had undergone endoscopic sinus surgery at least one year prior to the study with complete data of tissue specimens, baseline blood routine test, nasal endoscopy and sinus computed tomography, were identified and recruited to assess disease control based on the criteria of a European Position Paper on Rhinosinusitis and Nasal Polyps 2012 (EPOS 2012). Multiple logistic regression model was used to determine the association between blood and tissue eosinophil numbers and risk of poor disease control by adjusting for demographics and comorbidities. Results: We broke down the cohort into 4 groups according to blood (0.3×109/L) and tissue (10%) eosinophils. The patients without eosinophilic inflammation represented the largest group (41.5%). The group with concordant blood and tissue eosinophilia represented the second largest (31.2%), and the patients with isolated tissue (15.3%) or blood (12.0%) eosinophilia were relatively rare. Multiple logistic regression models found blood eosinophil count and tissue eosinophil percentage were independently associated with increased risk for poor disease control after adjustments for covariates related to poor treatment outcome. Furthermore, subjects with concordant blood and tissue eosinophilia had a higher risk for poor disease control than those with isolated blood or tissue eosinophilia. Conclusion: Concordant blood and tissue eosinophilia relates to a higher likelihood of poor disease control than isolated blood or tissue eosinophilia after adjustment of potential confounders in nasal polyp patients.

Effects of energy-restricted feeding during rearing on sexual maturation and reproductive performance of Rugao layer breeders.

The aim of this study was to assess the effects of energy-restricted feeding during rearing on the sexual maturation and reproductive performance of Rugao layer breeders. A total of 2,400 8-wk-old Rugao layer breeders were randomly assigned to one of 5 groups (480 pullets per group) with eight replicates and were fed one of 5 diets that were nutritionally similar with the exception of apparent metabolizable energy corrected for nitrogen (AMEn) content (2,850, 2,750, 2,650, 2,550, and 2,450 kcal AMEn/kg) from 8 to 18 wks of age. The daily amount of feed was restricted to the absolute quantity of the diet consumed by laying hens fed 2,850 kcal AMEn per kg diet ad libitum (control). From 18 to 52 wks of age, all hens were fed basal diets ad libitum. The body weight of layer breeders at 18 wks of age decreased linearly with increasing energy restriction (P < 0.001), but caught up within 3 wks of ad libitum feeding (P = 0.290). The coefficient of variation of the body weight of the hens at 18, 21, and 24 wks of age decreased linearly (P = 0.010, 0.025, and 0.041, respectively) with increasing energy restriction during rearing. Energy-restricted feeding delayed sexual organ development at 18, 20, and 22 wks of age, including the number of large yellow follicles, oviduct length, oviduct length index, oviduct index, and ovary stroma index (P < 0.05), and delayed sexual maturity, including the age at laying the first egg and the age at 5% and 50% egg production (P = 0.042, 0.004, and 0.029, respectively). Consequently, egg number from 5% to 50% egg production decreased linearly as the degree of energy restriction increased (P = 0.001) and egg production of hens in the energy-restricted feeding groups was lower than that of hens in the ad libitum feeding group (6.36, 6.43, 6.4, and 4.61% vs. 14.29%; P < 0.05) from 18 to 20 wks of age. Furthermore, egg weight increased linearly as energy restriction increased (P < 0.001) and laying hens in the most severe energy-restricted feeding group had more setting eggs (normal eggs weighing >40 g) than hens in the ad libitum feeding and lighter energy-restricted feeding groups (149.57 vs. 144.34, 142.66, 143.63, and 141.78; P < 0.05). No significant differences were observed in fertility, hatchability of fertile eggs, and hatchability of setting eggs (P = 0.381, 0.790, and 0.605, respectively). In conclusion, moderate energy restriction (85.97%, 2,450 vs. 2,850 kcal AMEn/kg) from 8 to 18 wks of age increased egg weight as well as the production of setting eggs in native layer breeders throughout the laying period, without adverse effects on productive performance from 18 to 52 wks of age, or fertility and hatchability at 52 wks of age.

Genome-wide association studies for mottled eggs in chickens using a high-density single-nucleotide polymorphism array.

Mottled eggs in layer chickens are gaining increasing attention because of the economic impact on the egg industry caused by the reduced sale value of commodity eggs. However, the genetic architecture underlying mottled eggs is not well understood. The genetic architecture underlying the mottled egg trait was investigated using genome-wide association studies (GWAS) by high-density arrays, using a total of 407 pink eggs and 799 blue eggs from an F2 resource population generated by crossing Dongxiang Blue-shelled and White Leghorn chickens. The mottled egg score in blue eggs was found to be higher than that in pink eggs. The single-nucleotide polymorphism heritability of mottled egg at laying day and storage for 7 days was 0.18 and 0.20, respectively. Bivariate GWAS provided 29 significant loci, mainly located on GGA2, GGA3, GGA8, GGA10, GGA15, GGA17, and GGA23, affecting mottled egg on laying day. Candidate genes RIMS2, SLC25A32, RIMBP2, VPS13B, and RGS3 were obtained for mottled eggshell by bivariate GWAS and gene annotation. Our findings provide new insights into the genetic architecture of mottled egg in hens, and demonstrate that a genomic selection method would be profitable for breeding out the mottled egg trait.

Efficacy evaluation of selenium-enriched yeast in laying hens: effects on performance, egg quality, organ development, and selenium deposition.

The aim of this study was to compare the dynamic changes of egg selenium (Se) deposition and deposition efficiency and to evaluate the efficacy of selenium-enriched yeast (SY) in laying hens over the 84 d feeding period after SY supplementation. A total of one thousand one hundred fifty-two 30-wk-old, Hy-Line Brown hens were randomly assigned to 1 of 6 groups (192 laying hens per group) with 6 replicates of 32 birds each, fed a basal diet (without Se supplementation), basal diet with 0.3 mg/kg of Se from sodium selenite (SS) or basal diets with 0.1, 0.2, 0.3, or 0.4 mg/kg of Se from SY, respectively. The results showed that the Se concentrations in the eggs and breasts from hens fed a SY-supplemented diet were significantly higher than those from hens fed a SS-supplemented diet or a basal diet (P < 0.001). There was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed a SY-supplemented diet and dietary Se supplementation on days 28, 56, and 84 (r2 = 0.931, 0.932, 0.976, P < 0.001; r2 = 0.946, 0.935, 0.976, P < 0.001), respectively. The Se deposition efficiency in whole eggs from hens fed a basal or SY-supplemented diet weresignificantly higher than those in eggs from hens fed a SS-supplemented diet on days 28, 56, and 84 (P < 0.001), respectively. In addition, there was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed SY-supplemented diet (r2 = 0.655, 0.779, 0.874, 0.781, P < 0.001; r2 = 0.666, 0.863, 0.944, 0.781, P < 0.001) or SS-supplemented diet (r2 = 0.363, P = 0.002; r2 = 0.440, P = 0.002) and number of feeding days. In conclusion, the organic Se from SY has higher bioavailability and deposition efficiency of Se in whole eggs as compared with inorganic Se from SS. The Se concentrations and Se deposition efficiency in the eggs increased from hens fed a SS- or SY-supplemented diet but decreased from hens fed a basal diet with the extension of the experimental duration. The results indicate that the dietary Se supplementation from SY should be limited to a maximum of 0.1 mg Se/kg complete feed when the eggs and meat produced from hens fed a SY-supplemented diet are used as food for humans directly, whereas up to 0.4 mg/kg organic Se from SY can be used to supplement the diets for laying hens when the products are used as raw materials for producing Se-enriched food.

Effects of high-dose selenium-enriched yeast on laying performance, egg quality, clinical blood parameters, organ development, and selenium deposition in laying hens.

Organic selenium (Se) supplementation from Se-enriched yeast (SY) has been advocated and approved for use in animal feeds by some nutritionists and researchers rather than inorganic Se from sodium selenite. However, there is little available safety data of SY in laying hens. A subchronic study was conducted to determine if high-dose SY affects the safety of hens. A total of 768, 30-wk-old, Hy-Line Brown hens were randomly assigned to 1 of 4 groups (192 laying hens per group) with 6 replicates of 32 birds each. After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 0.3, 1.5, or 3.0 mg/kg Se from SY for 12 wk. Throughout the study period, clinical observations and laying performance were measured. The hematological and chemical parameters of blood samples and the Se concentration in eggs were examined after SY supplementation for 4, 8, and 12 wk, and the egg quality was measured after 12 wk. At the end of the study, full post-mortem examinations were conducted: breast Se concentrations were measured, visceral, and reproductive organs were weighed, and specified tissues were collected for subsequent histological examinations. Although the Se concentrations in the eggs and breast meat from hens fed 3.0 mg/kg of Se from SY were 1036.73% and 2127.93% higher (P < 0.001) than those from hens fed a basal diet after 12 wk, no treatment-related changes of toxicological significance were observed. Therefore, up to 3 mg/kg organic Se from SY can be used to supplement the diets for laying hens without adverse effects following 84-d administration.

[Effect of CD4(+) T cell surface CD1d molecules on progression of multiple sclerosis in mouse experimental autoimmune encephalomyelitis model].

Objective: To investigate the effect of CD1d molecules on the surface of CD4(+) T cells on the progression of experimental autoimmune encephalomyelitis (experimental, allergic, encephalomyelitis, EAE) mouse models. Methods: EAE model of C57BL/6 mice was established, Splenic cells were isolated at different stages of the progression of the disease.The proportion of CD1d(+) cells on the surface of activated and non activated CD4(+) T cells was detected by flow cytometry. Results: The proportion of CD1d(+) cells in the control group (normal group and complete Freund's adjuvant (CFA) group), in the peak and recovery period of disease in the EAE group were compared.The proportion of CD1d(+) cells in the control group was (8.98 ±0.36)%, and the proportion of CD1d(+) cells in the peak and recovery period of disease in the EAE group were respectively (2.14±0.15)% and (13.80±0.84)%.The differences were statistically significant difference (P<0.05). Conclusion: The trend of the proportion of CD4(+) T cells expressing CD1d molecules during the course of EAE pathogenesis is verified, which lays a foundation for further study on the interaction between CD4(+) T cells and NKT cells in the progression of EAE models.

Comparison of dynamic change of egg selenium deposition after feeding sodium selenite or selenium-enriched yeast.

The aim of this study was to compare the dynamic change of egg selenium (Se) deposition after sodium selenite (SS) or selenium-enriched yeast (SY) supplementation for 1, 3, 5, 7, 14, 21, 28, 56, and 84 d. A total of 576 32-wk-old Hy-Line Brown laying hens were randomly assigned to 3 groups (192 laying hens per group) with 6 replicates, and fed a basal diet (without Se supplementation) or basal diets with 0.3 mg/kg of Se from SS or 0.3 mg/kg of Se from SY, respectively. The results showed that the Se concentrations in the eggs from hens fed a SY-supplemented diet were significantly higher (P < 0.001) than those from hens fed a SS-supplemented diet or a basal diet after 3 d. And the Se concentrations in the eggs from hens fed a SS-supplemented diet were significantly higher (P < 0.001) than those from hens fed a basal diet after 14 d. There was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed a SY-supplemented diet (r2 = 0.782, P < 0.001; r2 = 0.837, P < 0.001) or SS-supplemented diet (r2 = 0.355, P < 0.001; r2 = 0.413, P < 0.001) and number of feeding days. The Se concentrations in the breasts from hens fed a SY-supplemented diet were 126.98% higher (P < 0.001) than those from hens fed a SS-supplemented diet, and were 299.44% higher (P < 0.001) than those from hens fed a basal diet after the 84-d feeding period. In conclusion, the dietary Se was gradually transferred into eggs with the extension of the experimental duration. The deposition rate of Se in the eggs from hens fed a SY-supplemented diet was much more rapid than that from hens fed a SS-supplemented diet, and the organic Se from SY had higher bioavailability as compared to inorganic Se from SS.

Non-communicable chronic diseases and timely breast cancer screening among women of the Eastern Caribbean Health Outcomes Research Network (ECHORN) Cohort Study.

PURPOSE: The Caribbean population faces a growing burden of multiple non-communicable chronic diseases (NCDs). Breast cancer is the leading cause of cancer death for women in the Caribbean. Given the substantial burden of NCDs across the region, cancer prevention and control strategies may need to be specifically tailored for people with multiple co-morbidities. Preventive screening, such as timely mammography, is essential but may be either facilitated or hampered by chronic disease control. The main objective of this study is to examine the relationship between a chronic disease and timely breast cancer screening. METHODS: We conducted a cross-sectional data analysis using baseline data from the Eastern Caribbean Health Outcomes Research Network (ECHORN) Cohort Study-ECS. Our independent variables were presence of chronic diseases (hypertension or diabetes), defined as having been told by a clinical provider. Our dependent variable was timely screening mammography, as defined by receipt of mammography within the past 2 years. We examined bivariate and multivariate associations of covariates and timely screening mammography. RESULTS: In our sample (n = 841), 52% reported timely screening mammography. Among those with timely screening, 50.8% reported having hypertension, and 22.3% reported having diabetes. In our bivariate analyses, both diabetes and hypertension were associated with timely screening mammography. In partially adjusted models, we found that women with diabetes were significantly more likely to report timely screening mammography than women without diabetes. In our fully adjusted models, the association was no longer significant. Having a usual source of healthcare and a woman's island of residence were significantly associated with timely screening mammography (p < 0.05). CONCLUSIONS: We found that half of eligible women received timely screening mammography. Diabetes and hypertension, though common, are not associated with timely screening mammography. Usual source of care remains an important factor to timely breast cancer screening.

Safety evaluation of daidzein in laying hens: Effects on laying performance, hatchability, egg quality, clinical blood parameters, and organ development.

Daidzein has become increasingly popular as a dietary supplement, particularly for postpeak-estrus animals, as a safe and natural alternative estrogen-like compound. However, there is little available safety data of daidzein in laying hens. A study was conducted to examine if high-dose daidzein affected the safety of hens, including mortality, laying performance, egg quality, hematological parameters, clinical chemical parameters, organ development parameters, and hatchability. A total of 2,448 42-wk-old Rugao laying hens were randomly assigned to 4 groups with 6 replicates of 102 birds each (612 laying hens per group). After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 10, 100, or 200 mg/kg of daidzein for 12 wk. The hatchability of setting eggs increased linearly with increasing dietary daidzein supplementation (P = 0.034), while the hatchability of fertile eggs also tended to increase linearly (P = 0.069). The red cell distribution width (RCDW) and coefficient variation of RCDW showed an increasing and then decreasing quadratic response to increasing dietary daidzein supplementation (P = 0.001 and 0.002, respectively). No statistically significant changes were observed in mortality, laying performance, egg quality, clinical chemistry parameters, or organ development parameters (P > 0.05). The magnitude of these hematological changes was such that they were considered to be of no toxicological significance. Therefore, a nominal daidzein concentration of 200 mg/kg is not expected to cause adverse effects following daily administration to laying hens for 84 d.

Sequencing of Gag/Env association with HIV genotyping resolution and HIV-related epidemiologic studies of HIV in China.

HIV genotyping has led to conflicting results between laboratories. Therefore, identifying the most accurate gene combinations to sequence remains a priority. Datasets of Chinese HIV subtypes based on several markers and deposited in PubMed, Metstr, CNKI, and VIP databases between 2000 and 2015 were studied. In total, 9177 cases of amplification-positive samples from 26 provinces of China were collected and used to classify HIV subtypes based on eight individual genes or a combination thereof. CRF01_AE, CRF07_BC, CRF08_BC and B were the prevalent HIV subtypes in China, accounting for 84.07% of all genotypes. Gag/Env sequencing classified a greater number of HIV subtypes compared to other genes or combination of gene fragments. The geographical distribution of Gag and Gag/Env genotypes was similar to that observed with all genetic markers. Further principal component analysis showed a significantly different geographical distribution pattern of HIV in China for HIV genotypes detected with Gag/Env, which was in line with the distribution of all HIV genotypes in China. Gag/Env sequences had the highest diversity of the eight markers studied, followed by Gag and Gag/Pol/Env; Pol/Env polymorphisms were the least divergent. Gag/Env can serve as a high-resolution marker for HIV genotyping.

Improved protocol for extracting genomic DNA from frozen formalin-fixed tissue resulting in high-quality whole mtDNA.

Formalin fixation and paraffin embedding is widely used for convenient and long-term storage of tumor tissue and precious sources to perform genetic studies. However, DNA fragmentation is one of the major flaws of genomic DNA isolation from formalin fixation tissues, which limits its further usage. Here, we present an improved method for isolating high-quality genomic DNA from formalin fixation tissue. We obtained high-quality genomic DNA of more than 20 kb from samples frozen for more than 2 years. Furthermore, to verify DNA quality, the whole mitochondrial DNA (mtDNA) genomes from the normal and tumor tissue of the same patient were successfully amplified with two overlapping PCR fragments comprising more than 8379 bp in length for each fragment. In addition, the whole genomes were sequenced with a 48-well based primer panel in order to avoid potential sequencing errors from artificial recombination, which was further confirmed with an mtDNA phylogenetic strategy. Our improved DNA extraction method from formalin fixation tissue and sequencing strategy for entire mtDNA genomes will generate unambiguous sequence analysis results for clinical samples.

Use of health care services by patients with psoriasis: a population-based study.

BACKGROUND: Although psoriasis is seldom life threatening, very few studies have compared differences in health care service use between patients with and without psoriasis. OBJECTIVES: To investigate differences in health care service use between patients with and without psoriasis. METHODS: Patient details and data on their use of health services were retrieved from the Taiwan Longitudinal Health Insurance Database 2000. We included 3649 patients with psoriasis and 3649 without it. Each patient was followed for a 1-year period to estimate their utilization of health care resources. Student t-tests were used to compare differences in health care services use between patients with and without psoriasis. RESULTS: For dermatology services, patients with psoriasis had significantly more outpatient visits (3·5 vs. 0·9), and higher outpatient and total costs (US$148·00 vs. US$12·20 and US$581·60 vs. US$347·20, respectively) than those without psoriasis. For nondermatology services, patients with psoriasis had more outpatient visits (21·3 vs. 17·6), and higher outpatient and total costs (US$904·60 vs. US$663·50 and US$1335·50 vs. US$998·30, respectively) than those without psoriasis. For overall health care service use, patients with psoriasis had significantly more outpatient visits (24·8 vs. 18·5; P < 0·01) and greater total costs (US$1917·10 vs. US$1345·60; P < 0·01) than those without psoriasis. This indicates that the total cost was about 1·4-fold greater for patients with psoriasis than those without it. CONCLUSIONS: Patients with psoriasis used health care services significantly more often than those without psoriasis.

Scabies increased the risk of chronic kidney disease: a 5-year follow-up study.

BACKGROUND: The most documented complication of scabies has been reported to be infection by group A streptococci, which has in turn been suggested to contribute to the development of glomerulonephritis. OBJECTIVE: This study aimed to investigate the risk of chronic kidney disease (CKD) subsequent to scabies utilizing a population-based dataset in Taiwan. METHODS: This retrospective matched-cohort study included 5071 subjects with scabies and 25 355 randomly selected comparison subjects. We individually tracked each subject for a 5-year period to identify those who subsequently received a diagnosis of CKD during the follow-up period. Stratified Cox proportional hazards regressions were performed to compute the hazard ratio (HR) of CKD during the 5-year follow-up period. RESULTS: The incidence rate of CKD during the 5-year follow-up period was 9.66 (8.51-10.93) per 1,000 person-years and 6.24 (5.82-6.69) per 1000 person-years for subjects with and without scabies respectively. The HR for CKD during the 5-year follow-up period for subjects with scabies was 1.34 (95% CI = 1.15-1.56) that of comparison subjects after adjusting for monthly income, hypertension, diabetes, obesity, stroke, coronary heart disease, chronic obstructive pulmonary disease, tobacco use disorder, hyperlipidemia and alcohol abuse during the 5-year follow-up period. Male subjects with scabies were 1.40 (95% CI = 1.14-1.71) times more likely than comparison subjects to suffer from subsequent CKD, and female study subjects were 1.27 (95% CI = 1.05-1.61) times more likely. CONCLUSIONS: We concluded that there was an increased risk for CKD among patients suffering from scabies.

Increased risk of pemphigoid following scabies: a population-based matched-cohort study.

BACKGROUND: No prior study has investigated the possibility that scabies patients may be at an increased risk for developing pemphigoid. OBJECTIVE: To evaluate the risk of pemphigoid following scabies during a 3-year follow-up period using a Taiwanese population-based claims database and taking clinical and demographic characteristics into consideration. METHODS: This investigation consisted of a study group of 6793 subjects with a diagnosis of scabies and 33 965 randomly selected subjects used as a comparison group. Each patient was tracked for 3 years following their index dates to identify those who received a subsequent diagnosis of pemphigoid. Stratified Cox proportional hazards regressions were used to compute the hazard ratio (HR) of pemphigoid during the 3-year follow-up period. RESULTS: Of the 40 758 subjects, 52 (0.13%) had received a diagnosis of pemphigoid during the 3-year follow-up period; 33 (0.49% of the study group) were from the study group and 19 (0.06% of the comparison group) were from the comparison group. Compared to subjects without scabies, the HR for pemphigoid for subjects with scabies was 5.93 within the 3-year follow-up period following the index date after adjusting for monthly income, hypertension, diabetes, obesity, psoriasis, stroke, dementia, Parkinson's disease, coronary heart disease, schizophrenia or bipolar disorder, and after censoring those that died during the follow-up period. CONCLUSIONS: This study detected an increased risk for pemphigoid among patients suffering from scabies. Physicians treating elderly patients with a history of scabies should be alert to the development of pemphigoid.