Protective Role of Kelulut Honey against Toxicity Effects of Polystyrene Microplastics on Morphology, Hormones, and Sex Steroid Receptor Expression in the Uterus of Rats.

BACKGROUND: Microplastics (MPs) are an emerging global pollutant. Previous studies have revealed that chronic exposure to MPs can affect animal and human reproductive health, particularly by impairing the reproductive system's normal functions, which may increase the risk of infertility in both males and females. Kelulut honey (KH), an excellent source of antioxidants, has been used to counteract the disruptive effects of Polystyrene microplastics (PS-MPs) in the rat uterus. Thus, this study aimed to investigate the potential protective effects of Kelulut honey against PS-MPs-induced uterine toxicity in pubertal rats. METHODS: Prepubertal female Sprague Dawley rats were divided into four groups (n = 8): (i) normal control group (NC: treated with deionized water), MPs-exposed group (M: exposed to PS-MPs at 2.5 mg/kg), (iii) Kelulut honey group (DM: pretreated with 1200 mg/kg of KH 30 minutes before they were administered with PS-MPs at 2.5 mg/kg), and (iv) Kelulut honey control group (DC: only treated with KH at 2.5 mg/kg). The rats were treated orally once daily for six consecutive weeks. RESULTS: Uterine abnormalities in PS-MPs-exposed rats were significantly improved after concurrent treatment with Kelulut honey. Morphology improvement was observed and luminal epithelial cells seemed thicker with more goblet cells, glandular cells had a more regular and circular shape, stromal cell increased in size, interstitial gaps between stromal cells expanded, and the myometrium layer was thicker. Kelulut honey treatment also effectively normalized the suppressive effect of PS-MPs on the expression and distribution of sex steroid receptors (ERα and ERß), as well as the level of serum gonadotropin (LH and FSH) and sex steroid (estradiol and progesterone) hormones. CONCLUSION: Kelulut honey can protect the female reproductive system against the disruptive effects of PS-MPs. The phytochemical properties of Kelulut honey might be responsible for these beneficial benefits. However, future studies are warranted to identify the mechanisms involved.

Effects of Bypass Fat on Buffalo Carcass Characteristics, Meat Nutrient Contents and Profitability.

The deposition and distribution of buffalo body fats play a vital role in the quality of the buffalo carcass and are of great commercial value, since the carcass quality influences the profitability and consumer acceptability of ruminant meat. The current study examined the effect a mixture of 4% bypass fat and 26% concentrate supplementations in buffalo basal diet had on both the carcass characteristics and the proximate and fatty acid composition in longissimus thoracis et lumborum (LTL), supraspinatus (SS) and semitendinosus (ST) muscles of Murrah cross and swamp buffaloes. In addition, profit and loss analyses were performed to determine the profitability. This study employed a completely randomized 2 × 2 factorial arrangement with two diets, two breeds and four replicates per treatment. A total of sixteen buffaloes (eight buffaloes per breed, bodyweight 98.64 ± 1.93 kg) were randomly assigned into two dietary groups. The first group was given Diet A, which consisted of 70% Brachiaria decumbens + 30% concentrate, whereas the second group was given Diet B, which consisted of 70% Brachiaria decumbens + 26% concentrate + 4% bypass fat. The buffaloes were fed for 730 days before slaughter. The results showed that supplemented bypass fat significantly (p < 0.05) increased the pre-slaughter weight, hot and cold carcass weights, meat:fat ratio, pH at 24 h, moisture and crude protein of LTL, ST and SS, the ether extract of LTL and ST and the meat fatty acid of C16:0, C16:1, C18:1, PUFA n-6/n-3 and total MUFA. The carcass yield and carcass fat percentages, the ash content in ST, the EE in the SS muscle and the meat fatty acid of C18:3, total PUFA n-3, UFA/SFA and PUFA/SFA were significantly (p < 0.05) decreased. Furthermore, Murrah cross showed a significantly (p < 0.05) higher pre-slaughter weight, hot and cold carcass weights, carcass bone percentage and total fatty acid, but a lower (p < 0.05) meat:bone ratio, ash of LTL and CP of LTL and ST when compared to swamp buffaloes. No significant changes were found in the proximate composition of different types of muscle, but the ST muscle revealed significantly high C14:0, C16:0 and C18:1, and the SS muscle had high C18:2 and total fatty acid (p < 0.05). Supplementing using bypass fat increased the cost of buffalo feeding but resulted in a higher revenue and net profit. In conclusion, the concentrate and bypass fat supplementations in the buffalo diet could alter the nutrient compositions of buffalo meat without a detrimental effect on carcass characteristics, leading to a higher profit.

Telomere Length, Apoptotic, and Inflammatory Genes: Novel Biomarkers of Gastrointestinal Tract Pathology and Meat Quality Traits in Chickens under Chronic Stress (Gallus gallus domesticus).

This study was designed to examine the potentials of telomere length, mitochondria, and acute phase protein genes as novel biomarkers of gastrointestinal (GI) tract pathologies and meat quality traits. Chickens were fed a diet containing corticosterone (CORT) for 4 weeks and records on body weight, telomere length, GI tract and muscle histopathological test, meat quality traits, mitochondria, and acute phase protein genes were obtained at weeks 4 and 6 of age. The body weight of CORT-fed chickens was significantly suppressed (p < 0.05). CORT significantly altered the GI tract and meat quality traits. The interaction effect of CORT and age on body weight, duodenum and ileum crypt depth, pH, and meat color was significant (p < 0.05). CORT significantly (p < 0.05) shortened buffy coat telomere length. UCP3 and COX6A1 were diversely and significantly expressed in the muscle, liver, and heart of the CORT-fed chicken. Significant expression of SAAL1 and CRP in the liver and hypothalamus of the CORT-fed chickens was observed at week 4 and 6. Therefore, telomere lengths, mitochondria, and acute phase protein genes could be used as novel biomarkers for GI tract pathologies and meat quality traits.

Telomere Length and Regulatory Genes as Novel Stress Biomarkers and Their Diversities in Broiler Chickens (Gallus gallus domesticus) Subjected to Corticosterone Feeding.

This study was designed to characterize telomere length and its regulatory genes and to evaluate their potential as well-being biomarkers. Chickens were fed a diet containing corticosterone (CORT) for 4 weeks and performances, organ weight, plasma CORT levels, telomere lengths and regulatory genes were measured and recorded. Body weights of CORT-fed chickens were significantly suppressed (p < 0.05), and organ weights and circulating CORT plasma levels (p < 0.05) were altered. Interaction effect of CORT and duration was significant (p < 0.05) on heart and liver telomere length. CORT significantly (p < 0.05) shortened the telomere length of the whole blood, muscle, liver and heart. The TRF1, chTERT, TELO2 and HSF1 were significantly (p < 0.05) upregulated in the liver and heart at week 4 although these genes and TERRA were downregulated in the muscles at weeks 2 and 4. Therefore, telomere lengths and their regulators are associated and diverse, so they can be used as novel biomarkers of stress in broiler chickens fed with CORT.

Effects of Concentrate and Bypass Fat Supplementations on Growth Performance, Blood Profile, and Rearing Cost of Feedlot Buffaloes.

This study investigates the effects of supplementation of the basal diet with concentrate and rumen bypass fat on the dry matter intake (DMI), growth performance, blood metabolites and hormonal changes, and the feeding cost of feedlot water buffaloes. Thirty-six healthy, three- to four-month-old male Murrah crossbred (n = 18) and Swamp (n = 18) buffaloes with a similar average initial body weight of 98.64 ± 1.93 kg were each randomly allocated into three dietary experimental groups. Buffaloes were fed with Diet A, which consisted of 100% Brachiaria decumbens, Diet B, consisting of 70% Brachiaria decumbens and 30% concentrate, and Diet C, consisting of 70% Brachiaria decumbens, 26% concentrate, and 4% rumen bypass fat for a period of 730 days. Feed intake was measured daily, while blood samples were collected for every eight months. Furthermore, body scores were noted prior to and at the end of the experimental period. The results showed that the average daily gain for buffaloes fed with Diet C was the highest. The DMI, BCS, FI, and FCR for the three groups showed significant (p < 0.05) differences, in the following order: Diet C > Diet B > Diet A. At the end of the two-year feeding trial, buffaloes fed with Diet B had significantly (p < 0.05) higher cholesterol levels than Diet A and Diet C. In addition, buffaloes fed with Diet C had significantly (p < 0.05) higher levels of serum total protein, growth hormone, and insulin-like growth factor-I hormone compared to Diet A and Diet B. On the other hand, buffaloes fed with Diet B and Diet C showed significant (p < 0.05) decrease in glucose levels. Supplemented diet improved the buffalos' weight gain to achieve the market weight in a shorter period of time, thus, giving farmers a greater return. In conclusion, concentrate and bypass fat supplementations in the diet of water buffaloes improved the growth performance without adverse effect on the blood metabolites, which enabled better farmer profitability.

Comparative Growth and Economic Performances between Indigenous Swamp and Murrah Crossbred Buffaloes in Malaysia.

This study was conducted to compare the growth and economic performances between Swamp and Murrah crossbred buffaloes. The records of 108 Swamp and 276 Murrah crossbred buffaloes born between January 2010 and December 2015 were used in this study. The farm was practicing an extensive grazing system without supplementation from January 2010 to December 2011 (pre-intervention) and a new implementation of supplement in the feeding regime from January 2012 to December 2015 (post-intervention). The birth, weaning, and body weight at three monthly intervals, number of calves born, and mortality rate of calves at different years and during pre- and post-intervention were analyzed using a general linear model procedure. The interventions in 2012 had a positive effect on increasing the number of calves born for both breeds, average birth weight, economic performance, and reduce mortality calf rate. As a result, the birth weight of Murrah crossbred buffaloes was higher (36.63 ± 0.50 kg) than Swamp buffaloes (34.69 ± 0.40 kg) (p < 0.05). The average pre-weaning daily weight gain for Swamp and Murrah crossbred buffaloes was 0.73 and 0.98 kg/day (p < 0.05), while the average post-weaning daily weight gain was 0.39 and 0.44 kg/day, respectively (p < 0.05). The Swamp and Murrah crossbred buffaloes achieved the targeted market weight of 250 kg at 18 and 15 months old, respectively, while the targeted breeding weight of 385 kg was achieved at 30 and 26 months old, respectively. In this farm, on average a total of 64 calves were born yearly, with the ratio number of born calves per number of mated dams recorded higher in Murrah crossbred buffaloes as compared to Swamp buffalo (0.64 vs. 0.37) (p < 0.05). Furthermore, the average number of calves born in the post-intervention period (January 2012-December 2015) was significantly higher than in the pre-intervention period (January 2010-December 2011), respectively (Swamp: 23 vs. 8 and Murrah crossbred: 53 vs. 31, respectively) (p < 0.05). Partial budget method was used to estimate the net gain or loss between the two breeds. The average annual revenue was 2304.14 MYR (566.13 USD) for Swamp buffaloes and 4531.50 MYR (1113.39 USD) for Murrah crossbred buffaloes. The average annual cost saving was 340.02 MYR (83.54 USD) for Swamp and 215.75 MYR (53.01 USD) for Murrah crossbred buffaloes. On the other hand, annual added cost was 84.95 MYR (20.87 USD) for Swamp and 96.76 MYR (23.77 USD) for Murrah crossbred buffaloes. Therefore, the annual net benefit was 2559.21 MYR (628.80 USD) for Swamp and 4650.49 MYR (1142.63 USD) for Murrah crossbred buffaloes. As a conclusion, this study had shown that the higher average daily weight gain contributes to better cost savings, as shown by the crossbred buffaloes.

Quantification of Cooking Method Effect on COP Content in Meat Types Using Triple Quadrupole GC-MS/MS.

A diet containing cholesterol is an essential component of biological function; however, cholesterol oxidation products (COPs) remain a major public health concern. This study investigated the effects of cooking methods (boiling and frying) on the production levels of COPs in processed foods. Samples, as represented by minced beef, chicken sausages, and fish fillets, were subjected to different cooking methods followed by COP extraction using a saponification method. Then, six common COPs, 5α-cholest, α-epoxy, ß-epoxy, 25-HC, triol, and 7-keto, were quantified by triple quadrupole gas chromatography-mass spectrometry (GS-MS/MS). A significantly high number of COPs were detected in minced meat, of which 7-keto and triol were detected as major oxidation products, followed by chicken sausages and fish fillets (p ≤ 0.05). Compared to boiling, frying generated significantly more COPs, specifically triol (0.001-0.004 mg/kg) and 7-keto (0.001-0.200 mg/kg), in all samples. Interestingly, cholesterol level was found to be slightly (but not significantly) decreased in heat-treated samples due to oxidation during cooking, producing a higher number of COPs. Notably, the fish fillets were found to produce the fewest COPs due to the presence of a low amount of cholesterol and unsaturated fatty acids. In conclusion, adapting boiling as a way of cooking and choosing the right type of meat could serve to reduce COPs in processed foods.

Prevalence of chromosome anomalies in a deer farm with fertility decline in Malaysia.

BACKGROUND: A number of factors are known to reduce fertility rate in animals and one of the important categories of such factors is chromosome anomalies. They can occur with or without causing phenotypic abnormalities on animals; in some cases, they may directly affect meiosis, gametogenesis and the viability of conceptus. In many instances, balanced structural rearrangements can be transmitted to offspring, affecting fertility in subsequent generations. AIM: This work investigated the occurrence of chromosome aberrations in Rusa timorensis, Rusa unicolor and Axis axis raised in a nucleus deer farm in Malaysia with a history of declining fertility of unknown origin. MATERIALS & METHODS: Blood samples were collected from 60 animals through venipuncture, cultured for 72 h and arrested at metaphase. SmartType® and Ideokar® software were used to karyotype the chromosomes. RESULTS: We found 15 out of the 60 animals screened from both sexes harbor some form of chromosome aberration. Chromosomal aberrations exist at the rate of 25% and may not be unconnected with the observed reduced fertility on the farm. Further investigations should be carried out, especially on the offspring of the studied animals to transmission of these aberrations. The animals that are confirmed to transmit the chromosomal aberrations should be culled to arrest the propagation of their abnormalities.

Correction: Effects of dietary n-6: n-3 polyunsaturated fatty acid ratios on meat quality, carcass characteristics, tissue fatty acid profiles, and expression of lipogenic genes in growing goats.

[This corrects the article DOI: 10.1371/journal.pone.0188369.].

Dose-response relationship of tryptophan with large neutral amino acids, and its impact on physiological responses in the chick model.

Tryptophan (Trp) has been associated with the regulation of several behavioral and physiological processes, through stimulation of serotonergic activity. Tryptophan utilization at the metabolic level is influenced by the competitive carrier system it shares with large neutral amino acids (LNAA). This study was carried out using meat-type chicken as a model, to investigate the dose response effects of Trp/LNAA on fear response (tonic immobility; TI) and hormonal responses, including corticosterone (CORT), serotonin (5-HT), triiodothyronine (T3) and thyroxine (T4). A total of 12 cages (48 birds) were assigned to each of the six experimental groups at 29-42 days of age. Experimental diets were formulated to have incremental levels of Trp/LNAA (0.025, 0.030, 0.035, 0.040, 0.045, and 0.050). The results revealed that, Trp/NAA had no significant effect on growth performance and TI of the birds. However, elevation of Trp/LNAA was concurred with a linear reduction in CORT (P < .0001, r2 = 0.819) and linear increases in 5-HT (P < .0001, r2 = 0.945), T3 (P = .0003, r2 = 0.403) and T4 (P < .0001, r2 = 0.937) levels. In conclusion, the results from the current study demonstrated that, although incremental levels of Trp/LNAA did not affect bird growth performance or fearfulness, it increased 5-HT, T3 and T4, and decreased CORT levels in a linear dose-dependent manner. Manipulation of Trp feeding levels could be applied to manage stressful conditions in birds.

Prevalence of the CYP2C19*2 (681 G>A), *3 (636 G>A) and *17 (­806 C>T) alleles among an Iranian population of different ethnicities.

Polymorphisms in the cytochrome P (CYP) 450 family may cause adverse drug responses in individuals. Cytochrome P450 2C19 (CYP2C19) is a member of the CYP family, where the presence of the 681 G>A, 636 G>A and 806 C>T polymorphisms result in the CYP2C19*2, CYP2C19*3 and CYP2C19*17 alleles, respectively. In the current study, the frequency of the CYP2C19*2, CYP2C19*3 and CYP2C19*17 alleles in an Iranian population cohort of different ethnicities were examined and then compared with previously published frequencies within other populations. Allelic and genotypic frequencies of the CYP2C19 alleles (*2, *3 and *17) were detected using polymerase chain reaction (PCR)­restriction fragment length polymorphism analysis, PCR­single­strand conformation polymorphism analysis and DNA sequencing from blood samples of 1,229 unrelated healthy individuals from different ethnicities within the Iranian population. The CYP2C19 allele frequencies among the Iranian population were 21.4, 1.7, and 27.1% for the CYP2C19*2, CYP2C19*3 and CYP2C19*17 alleles, respectively. The frequency of the homozygous A/A variant of the CYP2C19*2 allele was significantly high and low in the Lur (P<0.001) and Caspian (P<0.001) ethnicities, respectively. However, the frequency of the homozygous A/A variant of the CYP2C19*3 allele was not detected in the Iranian cohort in the current study. The frequency of the heterozygous G/A variant of the CYP2C19*3 allele had the significantly highest and lowest frequency in the Fars (P<0.001) and Lur (P<0.001) groups, respectively. The allele frequency of the homozygous T/T variant of the CYP2C19*17 allele was significantly high in the Caspian (P<0.001) and low in the Kurd (P<0.05) groups. The frequency of the CYP2C19 alleles involved in drug metabolism, may improve the clinical understanding of the ethnic differences in drug responses, resulting in the advancement of the personalized medicine among the different ethnicities within the Iranian population.

Dietary Euphorbia hirta Extract Improved the Resistance of Sharptooth Catfish Clarias gariepinus to Aeromonas hydrophila.

Aqueous and methanol extracts of lemon Citrus limon peel, Euphorbia hirta (aerial parts), and fenugreek Trigonella foenum-graecum seeds were tested for their in vitro antimicrobial activities against the bacterium Aeromonas hydrophila. A swab paper disk method showed that the methanol extract of E. hirta (EHE) had the largest inhibition zone and the lowest minimal inhibitory concentration compared to all other herbal extracts. Based on these results, EHE was included in the diets of Sharptooth Catfish Clarias gariepinus at 0 (control), 2, 5, or 7 g/kg of diet (experiment 1). Each treatment was conducted in triplicate, with 30 fish (mean weight ± SE = 9.4 ± 0.4 g) in each replicate. After 30 d, the growth, feed intake, hepatosomatic index (HSI), and plasma biochemical parameters were measured. With a separate batch of Sharptooth Catfish, the efficacy of the EHE diets in conferring fish resistance to A. hydrophila over 30 d was compared to that of a diet containing oxytetracycline (OTC; experiment 2). Six treatments were conducted in triplicate groups of 30 fish (mean weight ± SE = 9.0 ± 0.3 g); the Control fish were fed the control diet and were not injected with A. hydrophila, while the Control-AH and OTC-AH groups were infected with A. hydrophila and were fed either the control diet or the diet containing OTC at 1 g/199 g. The other three treatments included fish that were injected with A. hydrophila but fed diets with increasing EHE at 2, 5, or 7 g/kg. Experiment 1 showed no change to growth, feeding efficiency, HSI, or plasma biochemical parameters. In experiment 2, however, fish that were fed dietary EHE at 5 g/kg had significantly lower mortality than the Control-AH group, with further resistance observed for fish fed EHE at 7 g/kg. Dietary OTC was more effective than EHE as a prophylactic to A. hydrophila infection in Sharptooth Catfish. Nevertheless, EHE can potentially be a valuable dietary supplement to improve the resistance of Sharptooth Catfish to A. hydrophila infection. Received May 3, 2017; accepted August 24, 2017.

Identification of a new mutation in an Iranian family with hereditary multiple osteochondromas.

BACKGROUND: Hereditary multiple osteochondromas (HMO), previously named hereditary multiple exostoses (HME), is an autosomal dominant skeletal disorder characterized by the growth of multiple osteochondromas and is associated with bony deformity, skeletal growth reduction, nerve compression, restriction of joint motion, and premature osteoarthrosis. HMO is genetically heterogeneous, localized on at least three chromosomal loci including 8q24.1 (EXT1), 11p11-p13 (EXT2), and 19p (EXT3). The median age of diagnosis is 3 years; almost all affected individuals are diagnosed by age 12. The risk for malignant degeneration to osteochondrosarcoma increases with age, although the lifetime risk of malignant degeneration is low (~1%). METHODS AND RESULTS: This study was performed on an Iranian family with nine affected individuals from three consecutive generations. Here, the proband was an affected woman who received genetic counseling prior to pregnancy. All exons of the three genes were examined in the proband using polymerase chain reaction and sequencing methods (the last member of this family is a male with severe deformities and lesions, especially around his large joints). Exon 4 of EXT1 (c.1235 G>A) was changed in affected individuals. This mutation alters tryptophan to a premature stop codon on amino acid position 412 (p.Trp412x). CONCLUSION: The outcome of this study has extended the genotypic spectrum of Iranian patients with HMO, revealing a way for improving detection and genetic counseling in carriers.

Dietary n-6:n-3 Fatty Acid Ratios Alter Rumen Fermentation Parameters and Microbial Populations in Goats.

Revealing the ruminal fermentation patterns and microbial populations as affected by dietary n-6:n-3 PUFA ratio would be useful for further clarifying the role of the rumen in the lipid metabolism of ruminants. The objective of the present study was to investigate the effects of dietary n-6:n-3 PUFA ratios on fermentation characteristics, fatty acid (FA) profiles, and microbial populations in the rumen of goats. A total of twenty-one goats were randomly assigned to three dietary treatments with different n-6:n-3 PUFA ratios of 2.27:1 (low ratio, LR), 5.01:1 (medium ratio, MR), and 10.38:1 (high ratio, HR). After 100 days of feeding, all goats were slaughtered. Dietary n-6:n-3 PUFA ratios had no effect (P > 0.05) on rumen pH and NH3N concentration. Goats fed HR diet had lower (P < 0.05) propionate and total volatile fatty acids and higher (P < 0.05) butyrate compared with those fed the MR and LR diets. The proportion of C18:0 decreased (P < 0.05) as dietary n-6:n-3 PUFA ratios increased. The proportions of C18:1 trans-11, C18:2n-6, cis-9 trans-11 CLA, and C20:4n-6 were greater in the HR goats compared with the MR and LR goats. Lowering dietary n-6:n-3 PUFA ratios enhanced (P < 0.05) the proportion of C18:3n-3 and total n-3 PUFA in the rumen fluid of goats. The populations of R. albus and R. flavefaciens decreased (P < 0.05) as the n-6:n-3 PUFA ratios increased in diet. Diet had no effect (P > 0.05) on the ruminal populations of F. succinogenes, total bacteria, methanogens, total protozoa, Entiodinium, and Holotrich. The population of B. fibrisolvens was lower (P < 0.05) in the LR goats compared with the MR and HR goats. It was concluded that HR would increase the concentration of cis-9 trans-11 CLA and C18:1 trans-11 in the rumen. However, LR whould decrease the B. fibrisolvens population, which is involved in the BH process in the rumen. Further research is needed to evaluate the potential role and contribution of rumen microbiome in the metabolism of FA in the rumen.

Conjugated linoleic acid: A potent fatty acid linked to animal and human health.

Conjugated linoleic acid (CLA) is a mixture of isomers of linoleic acid (C18:2 n-6), which is mostly found in the ruminant meat and dairy products. The CLA is known to have many potential health benefits, and considered a potent powerful fatty acid, which is linked to animal and human health. The present work aims to discuss the source and production, mechanism of action, and effects of CLA on humans, poultry, and ruminants by reviewing the recent studies carried out on CLA. Despite most of the recent studies indicating beneficial effects of CLA on improving body weight control parameters, its effects on reducing risk factors of cardiovascular diseases (CVD), inflammation, blood glucose, and insulin are still controversial, and need to be further studied in different hosts.

Manipulation of Rumen Microbial Fermentation by Polyphenol Rich Solvent Fractions from Papaya Leaf to Reduce Green-House Gas Methane and Biohydrogenation of C18 PUFA.

Different solvents (hexane, chloroform, ethyl acetate, butanol, and water) were used to identify the effect of papaya leaf (PL) fractions (PLFs) on ruminal biohydrogenation (BH) and ruminal methanogenesis in an in vitro study. PLFs at a concentration of 0 (control, CON) and 15 mg/250 mg dry matter (DM) were mixed with 30 mL of buffered rumen fluid and were incubated for 24 h. Methane (CH4) production (mL/250 mg DM) was the highest (P < 0.05) for CON (7.65) and lowest for the chloroform fraction (5.41) compared to those of other PLFs at 24 h of incubation. Acetate to propionate ratio was the lowest for PLFs compared to that of CON. Supplementation of the diet with PLFs significantly (P < 0.05) decreased the rate of BH of C18:1n-9 (oleic acid; OA), C18:2n-6 (linoleic acid; LA), and C18:3n-3 (α-linolenic acid; LNA) compared to that of CON after 24 h of incubation. Real time PCR indicated that total protozoa and total methanogen population in PLFs decreased (P < 0.05) compared to those of CON.

Association of nuclear and mitochondrial genes with audiological examinations in Iranian patients with nonaminoglycoside antibiotics-induced hearing loss.

Mitochondrial DNA mutations play an important role in causing sensorineural hearing loss. The purpose of this study was to determine the association of the mitochondrial genes RNR1, MT-TL1, and ND1 as well as the nuclear genes GJB2 and GJB6 with audiological examinations in nonfamilial Iranians with cochlear implants, using polymerase chain reaction, DNA sequencing, and RNA secondary structure analysis. We found that there were no novel mutations in the mitochondrial gene 12S rRNA (MT-RNR1) in patients with and without GJB2 mutation (GJB2(+) and GJB2(-), respectively), but a total of six polymorphisms were found. No mutations were observed in tRNA(Leu) (() (UUR) ()) (MT-TL1). Furthermore, eight polymorphisms were found in the mitochondrial ND1 gene. Additionally, no mutations were observed in the nuclear GJB6 gene in patients in the GJB2(-) and GJB2(+) groups. The speech intelligibility rating and category of auditory perception tests were statistically assessed in patients in the GJB2(-) and GJB2(+) groups. The results indicated that there was a significant difference (P<0.05) between the categories of auditory perception score in the GJB2(-) group compared to that in the GJB2(+) group. Successful cochlear implantation was observed among individuals with GJB2 mutations (GJB2(+)) and mitochondrial polymorphisms compared to those without GJB2 mutations (GJB2(-)). In conclusion, the outcome of this study suggests that variation in the mitochondrial and nuclear genes may influence the penetrance of deafness. Therefore, further genetic and functional studies are required to help patients in making the best choice for cochlear implants.

Prevalence of the rs7903146C>T polymorphism in TCF7L2 gene for prediction of type 2 diabetes risk among Iranians of different ethnicities.

BACKGROUND: Pharmacogenetics is the study of genetic polymorphisms affecting responses to drug therapy. The common rs7903146 (C>T) polymorphism of the TCF7L2 gene has recently been associated with type 2 diabetes (T2D). In this study, prevalence of the rs7903146 (C>T) polymorphism in the TCF7L2 gene for prediction of T2D risk was examined in an Iranian population of different ethnicities. METHODS: The prevalence of rs7903146 (C>T) and the predicted phenotypes, including extensive metabolizers, intermediate metabolizers, and poor metabolizers were investigated in blood samples of 300 unrelated healthy individuals in an Iranian population, including Fars, Turk, Lure, and Kurd, using polymerase chain reaction restriction fragment length polymorphism and direct genomic DNA sequencing. RESULTS: The homozygous wild-type (C/C), heterozygous (C/T), and homozygous (T/T) allelic frequencies of rs7903146 (C>T) in the TCF7L2 gene were 29% (extensive metabolizers), 66.34% (intermediate metabolizers), and 4.66% (poor metabolizers), respectively. The C/C, C/T, and T/T genotypic frequencies of the rs7903146 (C>T) allele were significantly different (P<0.01) among Iranians of different ethnicities. The frequency of the homozygous T/T variant of the rs7903146 (C>T) allele was significantly low in the Lure (P<0.01) and high in the Fars (P<0.001) ethnicities. Additionally, the frequency of the T/T variant of the rs7903146 (C>T) allele in the South of Iran was the highest (P<0.04), while the East of Iran had the lowest frequency (P<0.01). CONCLUSION: The prediction of rs7903146 (C>T) is required in drug research and routine treatment, where the information would be helpful for clinicians to optimize therapy and adverse drug reactions and predict drug response in individuals at risk of T2D.

Does Short-Term Dietary Omega-3 Fatty Acid Supplementation Influence Brain Hippocampus Gene Expression of Zinc Transporter-3?

Dietary omega-3 fatty acids have been recognized to improve brain cognitive function. Deficiency leads to dysfunctional zinc metabolism associated with learning and memory impairment. The objective of this study is to explore the effect of short-term dietary omega-3 fatty acids on hippocampus gene expression at the molecular level in relation to spatial recognition memory in mice. A total of 24 male BALB/c mice were randomly divided into four groups and fed a standard pellet as a control group (CTL, n = 6), standard pellet added with 10% (w/w) fish oil (FO, n = 6), 10% (w/w) soybean oil (SO, n = 6) and 10% (w/w) butter (BT, n = 6). After 3 weeks on the treatment diets, spatial-recognition memory was tested on a Y-maze. The hippocampus gene expression was determined using a real-time PCR. The results showed that 3 weeks of dietary omega-3 fatty acid supplementation improved cognitive performance along with the up-regulation of α-synuclein, calmodulin and transthyretin genes expression. In addition, dietary omega-3 fatty acid deficiency increased the level of ZnT3 gene and subsequently reduced cognitive performance in mice. These results indicate that the increased the ZnT3 levels caused by the deficiency of omega-3 fatty acids produced an abnormal zinc metabolism that in turn impaired the brain cognitive performance in mice.

Prevalence of the CYP2D6*10 (C100T), *4 (G1846A), and *14 (G1758A) alleles among Iranians of different ethnicities.

The presence of polymorphisms in the CYP2D6 gene may modulate enzyme level and activity, thereby affecting individual responses to pharmacological treatment. Here, we compared the prevalence of the CYP2D6*10, *4, and 14* alleles in an Iranian population of different ethnicities with those of other populations. Allele and genotype frequency distributions of CYP2D6*10 variants and predicted phenotypes including extensive metabolizers, intermediate metabolizers, and poor metabolizers were analysed in blood samples of 300 unrelated healthy individuals in an Iranian population using polymerase chain reaction (PCR)-restriction fragment length polymorphism, PCR-single-strand conformation polymorphism, and direct genomic DNA sequencing. The CYP2D6*4 (G1846A) and *14 (G1758A) allelic frequencies were not detected in different ethnicities, demonstrating the absence of a significant contribution of these alleles in Iranian populations. However, the T/T, C/T, and C/C genotype frequencies of the CYP2D6*10 allele were significantly different (P<0.01) in all Iranian ethnic groups. Additionally, the frequency of the homozygous T/T variant of the CYP2D6*10 allele was significantly high in the Lure (P<0.017) and low in the Kurd (P<0.002) ethnicities. The frequency of the T/T variant of the CYP2D6*10 allele in central Iran was the highest (P<0.001), while the south of Iran had the lowest frequency (P<0.001). The frequency of the C/T variant of the CYP2D6*10 allele was significantly a bit high (P<0.001) in females compare to males, while the frequencies of the T/T variant in females is similar to males, which are 24.4% and 24.3%, respectively. In contrast to absence of the CYP2D6*4 (G1846A) and *14 (G1758A) alleles in Iranian populations of different ethnicities, the prediction of the CYP2D6*10 allele is required in drug research and routine treatment, where the information would be helpful for clinicians to optimize therapy or identify persons at risk of adverse drug reactions before clinical trials. Approximately 39.3% of subjects (24.3% homozygous T/T CYP2D6*10 as poor metabolizers and 15% heterozygous C/T CYP2D6*10 as intermediate metabolizers) had this allele; therefore, the harmful effects of drugs are relatively common among Iranians.