Regulatory impacts of PPARGC1A gene expression on milk production and cellular metabolism in buffalo mammary epithelial cells.

The PPARGC1A gene plays a fundamental role in regulating cellular energy metabolism, including adaptive thermogenesis, mitochondrial biogenesis, adipogenesis, gluconeogenesis, and glucose/fatty acid metabolism. In a previous study, our group investigated seven SNPs in Mediterranean buffalo associated with milk production traits, and the current study builds on this research by exploring the regulatory influences of the PPARGC1A gene in buffalo mammary epithelial cells (BuMECs). Our findings revealed that knockdown of PPARGC1A gene expression significantly affected the growth of BuMECs, including proliferation, cell cycle, and apoptosis. Additionally, we observed downregulated triglyceride secretion after PPARGC1A knockdown. Furthermore, the critical genes related to milk production, including the STATS, BAD, P53, SREBF1, and XDH genes were upregulated after RNAi, while the FABP3 gene, was downregulated. Moreover, Silencing the PPARGC1A gene led to a significant downregulation of ß-casein synthesis in BuMECs. Our study provides evidence of the importance of the PPARGC1A gene in regulating cell growth, lipid, and protein metabolism in the buffalo mammary gland. In light of our previous research, the current study underscores the potential of this gene for improving milk production efficiency and overall dairy productivity in buffalo populations.

Analysis of tightly-coupled dipole phased array antennas with metasurface superstrate.

Tightly coupled dipole arrays, including connected arrays, and capacitively coupled arrays, are one of the best solutions for wideband phased array antenna designs. However, to increase bandwidth and maximum scan angle, we can use a metasurface superstrate. We propose an analytical model to compute the scan impedance of a tightly coupled dipole array loaded with a metasurface. This analytical model helps us to simplify the calculation of the scan impedance of the array and speeds up the design process of these array antennas. It is shown that the metasurface superstrate improves the bandwidth and beam scanning angles of the array. Using the proposed general transmission line model, the computation of the scan impedance of tightly coupled dipole arrays is done very fast with minimum error. The semi-analytical model is about 7 times faster than a full-wave simulation.

Evolutionary analysis of buffalo sterol regulatory element-binding factor (SREBF) family genes and their affection on milk traits.

The sterol regulatory element-binding factor (SREBF) genes are a vital group of proteins binding to the sterol regulatory element 1 (SRE-1) regulating the synthesis of fatty acid. Two potential candidate genes (SREBF1 and SREBF2) have been identified as affecting milk traits. This study aims to identify the SREBF family of genes and find candidate markers or SREBF genes influencing lactation production in buffalo. A genome-wide study was performed and identified seven SREBF genes randomly distributed on 7 chromosomes and 24 protein isoforms in buffalos. The SREBF family of genes were also characterized in cattle, goat, sheep and horse, and using these all-protein sequences, a phylogenetic tree was built. The SREBF family genes were homologous between each other in the five livestock. Eight single nucleotide polymorphisms (SNPs) within or near the SREBF genes in the buffalo genome were identified and at least one milk production trait was associated with three of the SNP. The expression of SREBF genes at different lactation stages in buffalo and cattle from published data were compared and the SREBF genes retained a high expression throughout lactation with the trend being the same for buffalo and cattle. These results provide valuable information for clarifying the evolutionary relationship of the SREBF family genes and determining the role of SREBF genes in the regulation of milk production in buffalo.

Association analysis between FASN genotype and milk traits in Mediterranean buffalo and its expression among different buffalo tissues.

Fatty acid synthase (FASN) is a multifunctional protein that catalyzes the synthesis of long-chain saturated fatty acid. In this study, we identified the single nucleotide polymorphisms (SNPs), and their association with milk traits in Mediterranean buffalo, and the expression of FASN gene in different tissues was measured. Nine SNPs (g.-1640G > A, g.-1099C > T, g.1095C > A, g.3221G > A, g.4762G > A, g.5299G > A, g.7164G > A, g.7272 T > C, and g.8927 T > C) were identified by DNA pooled sequencing and then genotyped. Seven identified SNPs except g.3221G > A and g.8927 T > C were found significantly associated with both fat and protein percentage, and also the g.7164G > A and g.8927 T > C had significant association with peak milk yield and protein percentage, respectively. One haplotype block was successfully constructed by linkage disequilibrium (LD) analysis and it showed a significant association with both fat percentage and protein percentage. Expression of FASN gene was found in almost all the buffalo tissues including mammary gland, heart, liver, spleen, lung, kidney, uterus, and ovary, and to be highest in lung and mammary gland. Our findings suggest that polymorphisms in the buffalo FASN gene are associated with milk production traits and can be used as a candidate gene for milk traits and marker-assisted selection in buffalo breeding program.

Genetic Association of PPARGC1A Gene Single Nucleotide Polymorphism with Milk Production Traits in Italian Mediterranean Buffalo.

PPARGC1A gene plays an important role in the activation of various important hormone receptors and transcriptional factors involved in the regulation of adaptive thermogenesis, gluconeogenesis, fiber-type switching in skeletal muscle, mitochondrial biogenesis, and adipogenesis, regulating the reproduction and proposed as a candidate gene for milk-related traits in cattle. This study identified polymorphisms in the PPARGC1A gene in Italian Mediterranean buffaloes and their associations to milk production and quality traits (lactation length, peak milk yield, fat and protein yield, and percentage). As a result, a total of seven SNPs (g.-78A>G, g.224651G>C, g.286986G>A, g.304050G>A, g.325647G>A, g.325817T>C, and g.325997G>A) were identified by DNA pooled sequencing. Analysis of productivity traits within the genotyped animals revealed that the g.286986G>A located at intron 4 was associated with milk production traits, but the g.325817T>C had no association with milk production. Polymorphisms in g.-78A>G was associated with peak milk yield and milk yield, while g.304050G>A and g.325997 G>A were associated with both milk yield and protein percentage. Our findings suggest that polymorphisms in the buffalo PPARGC1A gene are associated with milk production traits and can be used as a candidate gene for milk traits and marker-assisted selection in the buffalo breeding program.

Adaptive Molecular Evolution of AKT3 Gene for Positive Diversifying Selection in Mammals.

The V-Akt Murine Thymoma Viral Oncogene Homolog 3 (AKT3) gene is of the serine/threonine-protein kinase family and influences the production of milk fats and cholesterol by acting on the sterol administrative area restricting protein (SREBP). The AKT3 gene is highly preserved in animals, and during lactation in cattle, its expression increases. The AKT3 gene is expressed in the digestive system, mammary gland, and immune cells. A phylogenetic investigation was performed to clarify the evolutionary role of AKT3, by maximum probability. The AKT3 gene sequence data of various mammalian species was evident even with animals undergoing breeding selection. From 39 mammalian species studied, there was a signal of positive diversifying selection with Hominidae at 13Q, 16G, 23R, 24P, 121P, 294K, 327V, 376L, 397K, 445T, and 471F among other codon sites of the AKT3 gene. These sites were codes for amino acids such as arginine, proline, lysine, and leucine indicating major roles for the function of immunological proteins, and in particular, the study highlighted the importance of changes in gene expression of AKT3 on immunity.

Effects of In Ovo Injection of Coenzyme Q10 on Hatchability, Subsequent Performance, and Immunity of Broiler Chickens.

Effects of in ovo injection of Q10 on hatchability, performance (feed intake (FI), body weight gain (BWG), feed/gain ratio (F/G)) traits, and immune status of Ross × Ross 308 broiler chicks, hatched from eggs laid by a 38-week-old breeder flock, were determined through 42 days after hatch. Eggs containing live embryos were injected in the amnion with 0.1 and 0.2 mL Q10 solution on day 18 of incubation. Two controls groups were included as sham and/or as an uninjected group. At 28 and 42 days of age, performance traits, serum enzyme activity, weights of immune organs, and serum antibody titer of viral diseases were determined. Results were shown that hatchability % increased by Q10 on average of 6.54% (P≤0.025) and body weight/egg weight after hatching increased up to 4.74% (P≤0.002), compared with uninjected and sham controls. Injection of Q10 at different levels led to significant increases (P≤0.001) in performance traits all over the rearing period (P<0.05). Weight of immune organs significantly improved compared to uninjected and sham controls (P<0.05). In addition, serum antibody titers of viral diseases as well as serum enzyme activity of AST, ALT, CAT, and SOD were significantly changed by Q10 treated groups than controls (P≤0.01). In conclusion, in ovo injection of Q10 at levels of 0.1 and 0.2 mL led to significant increases in hatchability%, internal egg characteristics, and performance parameters as well as serum enzyme activity, weight of immune organs, and serum antibody titer of ND, AI, and IBD diseases.

The effect of trabeculectomy on contrast sensitivity, corneal topography and aberrations.

PURPOSE: To investigate the effect of trabeculectomy on corneal topography, corneal sensitivity and aberrations. METHODS: Twenty-four eyes of 24 subjects with open-angle glaucoma who required glaucoma filtration surgery were assessed. The evaluation of corneal topography, corneal sensitivity and aberrations were done before the trabeculectomy procedure, 1 week and 1 month after the trabeculectomy. RESULTS: There were significant differences in cylindrical power (P = 0.02), contrast sensitivity at 12 cycle/degree spatial frequency (P = 0.04) as well as high order aberration (P = 0.04) and high order without spherical component (P = 0.02) following trabeculectomy. However, significant differences were found for keratometric results and Fourier index in 3 and 6 mm pupil diameters between pre- and post-trabeculectomy (P > 0.05). CONCLUSIONS: According to the findings of the current study, trabeculectomy affects contrast sensitivity at 12 cycle/degree spatial frequency, higher-order aberration and higher order without spherical component aberration 1 month after trabeculectomy. Being knowledgeable about these changes may lead to some advancement in post-surgical management of patients particularly in early stages following trabeculectomy.

Genetic variants in the promoter region of the KLF3 gene associated with fat deposition in Qinchuan cattle.

Krüppel-like factor 3 (KLF3), a member of the Krüppel-like factor (KLF) family, plays an important role in adipogenesis and lipid metabolism. The aim of this study was to investigate whether KLF3 could be used as a candidate gene in the breeding of cattle. The expression pattern of bovine KLF3 gene revealed that it was highly expressed in abdominal fat and perirenal fat. Using DNA sequencing, three single nucleotide polymorphisms (SNPs) within the promoter regions of KLF3 gene were identified in 448 Qinchuan cattle, which are located in the recognition sequences of 11 transcription factors and the four haplotypes representing four potential different compositions of polymorphic potential cis-acting elements. Association analysis results indicated that individuals with the Hap7/7 diplotype showed higher (P < 0.05) intramuscular fat content (IFC) than those with H7/8. In addition, the H7 haplotype had much higher (P < 0.05) transcriptional activity than the H8 haplotype, consistent with the association analysis. We speculated that polymorphisms in transcription factor binding sites of the KLF3 promoter region affected transcriptional activity of KLF3, which subsequently influence intramuscular fat content in Qinchuan cattle and KLF3 gene could be used as molecular markers for fat deposition traits using early marker-assisted selection (MAS) of Qinchuan cattle breeding in the future.

KLF15 promotes transcription of KLF3 gene in bovine adipocytes.

The Krüppel-like factors (KLF) family plays an important role in adipogenesis, which is subject to internal hierarchical regulation. KLF3 is a member of KLF family, mainly responsible for adipocyte differentiation and fat deposition. However, the transcriptional regulation of bovine KLF3 gene and its relationship with KLF15 gene remains unclear during bovine adipogenesis. Here, we report that the expression pattern of KLF3 and KLF15 genes during bovine adipogenesis, when KLF15 gene was overexpressed through adenoviral vector (Ad-KLF15) in bovine adipocytes the expression level of KLF3 gene was increased, similarly when KLF15 was down regulated through siRNA the expression level of KLF3 was also reduced. To explore the transcriptional regulation of bovine KLF3 gene and its relationship with KLF15, serial deletion constructs of the 5'flanking region of bovine KLF3gene revealed through dual-luciferase reporter assay that the core promoter is located in -264 to -76 regions. The most proximal GGGG element in the promoter of the bovine KLF3 gene (located in -264 to -76 regions) is required for promotion by KLF15. Electrophoretic mobility shift (EMSA) and chromatin immunoprecipitation (ChIP) assays further confirmed that KLF15 gene binds to the KLF3 gene core promoter region in bovine adipocytes. These findings conclude that KLF15 promotes the transcriptional activity of KLF3 in bovine adipocytes. This mechanism to provides a new direction for further study of adipogenesis by internal regulation of members within KLF family.

Polymorphism in promoter of SIX4 gene shows association with its transcription and body measurement traits in Qinchuan cattle.

The sine oculis homeobox homolog 4 (SIX4) gene belongs to the SIX gene family, which plays a critical role in muscle regeneration and early stages of ontogeny. This study aimed to detect promoter variations of bovine SIX4 genes in Qinchuan cattle, and to evaluate the effect of transcription regulations and body measurement traits. Quantitative real-time PCR (qPCR) results showed that the mRNA expression levels of SIX4 gene were found significantly highest in longissimus thoracis tissue and individual before attaining the stage of physiological maturity. Using sequencing technology on a total of 428 Qinchuan cattle, seven single nucleotide polymorphisms (SNPs) were identified in the promoter region of SIX4, and seven haplotypes representing 18 potential transcription factor compositions of polymorphic potential cis-acting elements. Association analysis indicated that the H3-H3 diplotype performed greater withers height, chest depth, chest circumference, back fat thickness and ultrasound loin muscle area (P < 0.05) than H5-H6, which were consistent with the promoter activity of Hap3 haplotype was higher than the Hap5 and Hap6 haplotype in vitro. These potential transcription factor information and combined genotypes H3-H3 of the SIX4 gene can be used as a molecular marker for selection of economic traits in Qinchuan cattle.

Evaluation of anterior chamber angle under dark and light conditions in angle closure glaucoma: An anterior segment OCT study.

AIM: To evaluate changes of nasal and temporal anterior chamber angle (ACA) in subjects with angle closure glaucoma using Spectralis AS-OCT (SAS-OCT) under dark and light conditions. METHODS: Based on dark-room gonioscopy, 24 subjects with open angles and 86 with narrow angles participated in this study. The nasal and temporal angle opening distance at 500 µm anterior to the scleral spur (AOD500), nasal and temporal ACA were measured using SAS-OCT in light and dark conditions. RESULT: In 2 groups, ACA and AOD500 in nasal and temporal quadrants were significantly greater in light compared to dark (all with p=0.000). The AOD500 and ACA were significantly higher in nasal than temporal in measured conditions for 2 groups except the ACA and AOD500 of normal group measured in light. The difference between nasal and temporal in dark (29.07 ± 65.71 µm for AOD500 and 5.7 ± 4.07° for ACA) was greater than light (24.86 ± 79.85 µm for AOD500 and 2.09 ± 7.21° for ACA) condition. But the difference was only significant for ACA (p=0.000). The correlation analysis showed a negative correlation between AOD500 and pupil diameter in temporal and nasal quadrants (both with p=0.000). While temporal AOD500 difference correlated with spherical equivalent, temporal and asal gonioscopy, nasal AOD correlated with IOP, temporal and nasal gonioscopy. CONCLUSIONS: Clinically important changes in ACA structure could be detected with SAS-OCT in nasal and temporal quadrants under different illumination intensity. The results could help in improvement of examination condition for better and more accurate assessment of individuals with angle closure glaucoma.

Prevalence of Refractive Errors in Students with and without Color Vision Deficiency.

PURPOSE: To evaluate refractive errors in school age children with color vision deficiency (CVD) and those with normal color vision (NCV) in order to make a better understanding of the emmetropization process. METHODS: A total of 4,400 primary school students aged 7-12 years were screened for color vision using Ishihara pseudoisochromatic color vision plate sets. Of these, 160 (3.6%) students had CVD. A total of 400 age- and sex-matched students with NCV were selected as controls. Refractive status was evaluated using objective cyclorefraction. RESULTS: The CVD group included 136 male (85%) and 24 female (15%) subjects with mean age of 10.1 ± 1.8 years. The NCV group comprised of 336 male (84%) and 64 female (16%) subjects with mean age of 10.5 ± 1.2 years. The prevalence of myopia (7.7% vs. 13.9%, P < 0.001) and hyperopia (41% vs. 57.4%, P = 0.03) was significantly lower in the CVD group. Furthermore, subjects with CVD subjects demonstrated a lower magnitude of refractive errors as compared to the CVD group (mean refractive error: +0.54 ± 0.19 D versus + 0.74 ± 1.12 D, P < 0.001). CONCLUSION: Although the lower prevalence of myopia in subjects with CVD group supports the role of longitudinal chromatic aberration in the development of refractive errors; the lower prevalence of hyperopia in this group is an opposing finding. Myopia is a multifactorial disorder and longitudinal chromatic aberration is not the only factor influencing the emmetropization process.

Efficacy of corneal tomography parameters and biomechanical characteristic in keratoconus detection.

AIM: To determine the efficacy of corneal thickness parameters and corneal biomechanical properties (CBPs) in discriminating between normal and keratoconus eyes. METHOD: After performing a comprehensive ophthalmic examination, 50 mild to moderate keratoconus and 50 age and sex matched myopic astigmatism eyes were prospectively included in the study. The corneal topographic maps and CBP were obtained by Pentacam and Ocular response analyser, respectively. Central corneal thickness (CCT), thinnest corneal thickness (TCT), corneal thickness (CT) and percentage thickness increase (PTI) at 1, 3 and 5mm from the thinnest point and corneal volume (CV) at 3, 5, 7 and 10 centred on thinnest point, corneal hysteresis (CH) and corneal resistance factor (CRF) were recorded. Independent t-test and receiver operating characteristic (ROC) were done with SPSS software (version 15.0, SPSS, Inc.). RESULTS: CCT, TCT, CT at 1, 3 and 5, CV at 3, 5, 7 and 10mm, CH and CRF were significantly lower in keratoconus eyes compared to controls (p<0.001). In addition, PTI at 1, 3 and 5mm from the thinnest point showed significantly higher values in keratoconus group. ROC analysis demonstrated good predictive accuracy for cut-off point values. However, the centrally located indices had higher predictive accuracy compared to the peripherally located indices. CONCLUSION: Although good sensitivity and specificity were found for the mentioned parameters, the centrally located indices had higher predictive accuracy compared to peripherally located indices. It is suggested to use a combination of corneal pachymetry together with CBP for more accurate detection of keratoconus.

Assessment of the anterior chamber parameters after laser iridotomy in primary angle close suspect using Pentacam and gonioscopy.

AIM: To evaluate the changes in the anterior segment parameters of the subjects with primary angle closure suspect (PACS) before and after laser iridotomy (LI) using the Pentacam and gonioscopy. METHODS: Forty-eight eyes of 48 PACS were included. Anterior chamber angle (ACA), central anterior chamber depth (ACD), anterior chamber volume (ACV) and central corneal thickness (CCT) were recorded from the Pentacam before and one month after LI. ACA was graded according to Shaffer classification using the Goldmann gonioscopy. RESULTS: ACA increased significantly from 25.59±4.41 to 26.46±4.33 degrees (P=0.009) and ACV changed from 85.97±16.07mm(3) to 99.25±15.83mm(3) (P=0.000). The changes in ACD, CCT and intraocular pressure were non-significant (P>0.05). Gonioscopy showed significant widening of the Shaffer angle in 4 quadrants (P<0.001). CONCLUSION: Pentacam can serve as the objective instrument in assessing the efficacy of LI.

Anterior segment changes following short-term reading and its correlation with corneal biomechanical characteristics.

PURPOSE: To investigate the changes in anterior segment following short-term reading and evaluate the correlation of such changes with corneal biomechanical characteristics (CBC). METHODS: Thirty-six right eyes of 36 healthy subjects were examined. Anterior segment parameters were measured using the Pentacam before and after 30 min of reading. Ocular Response Analyzer was used to record CBC after reading. The following were recorded: central corneal thickness (CCT), central corneal power (CCP), superior corneal power (SCP), inferior corneal power (ICP), anterior chamber depth (ACD), anterior chamber volume (ACV), anterior chamber angle (ACA), corneal hysteresis (CH) and corneal resistance factor (CRF). Statistical analysis was performed with the paired student t-test and Pearson correlation test in SPSS 16. RESULTS: There were statistically significant decreases in CCP, SCP, ACD and ACV values following reading (p < 0.05). Our results showed a statistically significant negative correlation between CH and changes in ICP (r = 0.36, p = 0.02). Significant negative correlations were also found between CRF and changes in ICP (r = 0.41, p = 0.01) and SCP (r = 0.34, p = 0.04). On the other hand, statistical analysis indicated no correlation between CBC and other studied parameters (p > 0.05). CONCLUSION: This study demonstrated significant changes in some anterior segment parameters after reading. Being knowledgeable about these changes may have important implications in high accuracy examinations such as pre-operative assessment of corneal refractive surgery candidates. This could also help researchers have a better understanding of the factors that may influence near work related development of refractive errors.

Anterior segment parameters in Indian young adults using the Pentacam.

To evaluate Pentacam-Scheimpflug imaging of anterior segment parameters in young Indian adults. In this prospective study 120 eyes of 60 normal Indian subjects with a mean age of 25.93 ± 6.58 years (range 17-39 years) were assessed by Pentacam. Main outcome measures were central corneal thickness (CCT), thinnest corneal thickness (TCT), apex corneal thickness (apex CT), peripheral corneal thickness at 2, 4, 6 and 8 mm from the thinnest point, location of the thinnest pachymetry, corneal volume (CV), anterior chamber depth (ACD), anterior chamber volume (ACV) and anterior chamber angle (ACA). Independent samples t test, dependent samples t test, ANOVA and Pearson correlation test were used for statistical analysis. The mean CCT, TCT, Apex CT and CV were 544.95 ± 35.42, 542 ± 35.19, 545.43 ± 35.45 and 61.64 ± 4.17 µm, respectively. There was a gradual increase in CT from the thinnest point to the periphery. The mean ACD was 3.14 ± 0.33 mm, mean ACV was 177.77 ± 29.02 mm(3), and mean ACA was 39.36° ± 5.42°. There was no significant difference between CCT, TCT and Apex CT. A significant positive correlation was found between CCT and peripheral CT and also between anterior chamber parameters. TCT was mainly located in the inferotemporal and superotemporal zone. No significant difference was found in parameters between the right and left eyes and also between genders. This study provided information about a wide range of parameters in the anterior segment of healthy Indian eyes. These results could be helpful in assessment of patients with corneal diseases, glaucoma and screening for refractive surgeries.

Anterior segment characteristics of keratoconus eyes in a sample of Asian population.

PURPOSE: To assess changes in anterior segment parameters of keratoconus eyes at different stages of the disease in a sample of the Asian population. METHODS: Files of 32 patients (48 eyes) diagnosed as clinical keratoconus were assessed and the following parameters noted: central corneal thickness (CCT), thinnest corneal thickness (TCT), location of thinnest pachymetry, anterior chamber depth (ACD) at the centre from posterior corneal surface, ACD at 1, 2 and 3mm inferior-paracentral, ACD at thinnest pachymetry, anterior chamber volume (ACV) and anterior chamber angle (ACA). For analysis, keratoconus eyes were classified into 3 subgroups according to mean keratometry readings (mild: K≤47.0D, moderate: 47.0

Corneal thickness and volume in subclinical and clinical keratoconus.

To evaluate corneal thickness and volume in subclinical and clinical keratoconus in Asian population with the aim of discriminating between normal and ectatic cornea. Eyes were placed into one of the following three groups: normal, subclinical, and mild-moderate keratoconus. Pentacam Scheimpflug imaging (Oculus Inc., Wetzlar, Germany) was performed for each participant to record thinnest corneal thickness, central corneal thickness, corneal volume (CV), peripheral corneal thickness (PCT) and percentage thickness increase (PTI) at 2, 4, 6, and 8 mm. The data were exported to SPSS for statistical analysis. Subjects comprised 52 normal, 15 subclinical keratoconus, and 32 mild-moderate clinical keratoconus eyes. Our results indicated that corneal thickness (CT) distribution, PTI, and CV in normal eyes were significantly different compared with subclinical and clinical keratoconus (P < .05). Overall, subclinical group exhibited lower CT distribution and volume, and higher PTI in comparison with normal eyes. However, they showed higher CT distribution and volume, and lower PTI compared with keratoconus group. In addition, there was a smaller change in PCT and PTI from the thinnest point of the cornea to the periphery. The results of the present study indicate that CT parameters and CV were significantly different in normal versus subclinical group and in normal versus keratoconus group. These findings could help clinicians to better discriminate between normal and ectatic cornea.