Deaddicta® for maintenance treatment of Opioid-dependence: A six-month follow-up.

Background: Opioid dependence, is one of the world's most critical health problems. Deaddicta is a herbal product considered an effective treatment for opioid addiction. Deaddicta's efficacy in the maintenance treatment of patients with opioid use disorder has recently been demonstrated through a double-blind randomized controlled trial (RCT). This study aimed to evaluate the permanence of Deaddicta's efficacy six months after the end of the maintenance treatment for opioid dependence. Methods: This study was performed following the previous RCT on the maintenance treatment of opioid addicts. Out of 41 participants who completed the study for three months in the previous research, 15 from the intervention group (Deaddicta capsules, 1500 mg/day) returned for follow-up. They all previously fulfilled the DSM-IV criteria for addiction, were aged 18 to 65, and had discontinued Deaddicta for six months. The outcome measures included addiction severity, depression and anxiety levels, and craving score. The scores of each parameter were compared in three phases: before intervention; after three months of intervention; and six months after the end of the study. Results: Depression, anxiety, and craving scores decreased six months after the end of the previous study. This decrease was significant in the craving score (P = 0.011). No significant increase was observed in the frequency of use. The regression analysis showed a negative relationship between craving and the progression of phases. Conclusion: The Deaddicta product may have desirable and effective properties in decreasing temptation and, as a result, the maintenance treatment of opioid dependence.

Antioxidant and antihyperlipidemic effects of hawthorn extract (Crataegus oxyacantha) in broiler chickens.

BACKGROUND: One of the main problems in the poultry industry is excess body fat, and the anti-fat effect of Cratagus extract has been confirmed in several studies. OBJECTIVES: The present study was carried out to investigate the effects of hawthorn extract (Crataegus oxyacantha) on growth performance, haematological variables and hepatic gene expression in broiler chickens reared at high altitude (2100 m). METHODS: A total of 225-day-old chicks (Ross 308) were randomly distributed into three treatments. Experimental treatments were prepared by adding 0.0, 0.2 and 0.4 mL of hawthorn extract per litre of consumption of water. RESULTS: The results showed that weight gain and feed conversion ratio were significantly improved and abdominal fat decreased by consumption of two levels of Crateagus extract when compared to the control (p < 0.05). Consumption of hawthorn extract decreased circulatory levels of malondialdehyde, triacylglycerol, total cholesterol and low-density lipoproteins cholesterol but increased ferric reducing antioxidant power and high-density lipoproteins cholesterol (p < 0.05). Hawthorn extract caused an up-regulation of catalase, superoxide dismutase1, glutathione peroxidase1 and peroxisome proliferator-activated receptor alpha but reduced the expression of key lipogenic enzymes (p < 0.05). CONCLUSIONS: Overall, consumption of 0.4 mL hawthorn extract per litre of drinking water, improved growth performance, suppressed lipogenesis and enhanced antioxidant response.

Canine sperm motility is associated with telomere shortening and changes in expression of shelterin genes.

BACKGROUND: Motion quality is a critical property for essential functions. Several endogenous and exogenous factors are involved in sperm motility. Here, we measured the relative telomere length and evaluated the gene expression of its binding-proteins, shelterin complex (TRF1, TRF2, RAP1, POT1, TIN2, and TPP1) in sperm of dogs using relative quantitative real-time PCR. We compared them between two sperm subpopulations with poor and good motion qualities (separated by swim-up method). Telomere shortening and alterations of shelterin gene expression result from ROS, genotoxic insults, and genetic predisposition. RESULTS: Sperm kinematic parameters were measured in two subpopulations and then telomeric index of each parameter was calculated. Telomeric index for linearity, VSL, VCL, STR, BCF, and ALH were significantly higher in sperms with good motion quality than in sperms with poor quality. We demonstrated that poor motion quality is associated with shorter telomere, higher expression of TRF2, POT1, and TIN2 genes, and lower expression of the RAP1 gene in dog sperm. The levels of TRF1 and TPP1 gene expression remained consistent despite variations in sperm quality and telomere length. CONCLUSION: Data provided evidence that there are considerable changes in gene expression of many shelterin components (TRF2, TIN2, POT1and RAP1) associated with shortening telomere in the spermatozoa with poor motion quality. Possibly, the poor motion quality is the result of defects in the shelterin complex and telomere length. Our data suggests a new approach in the semen assessment and etiologic investigations of subfertility or infertility in male animals.

Neuroprotective anticonvulsant and anxiolytic effects of octreotide in wistar rats.

Somatostatin interneurons exhibited anti-epileptic activity. As a result, somatostatin agonists appear to be a promising target for antiepileptic drug development (AEDs). In this regard, we investigated the effects of octreotide, a somatostatin analog, on pentylenetetrazol (PTZ)-induced seizures in male Wistar rats. Animals were given octreotide at doses of 50 or 100 µg/kg for seven days. The anxiolytic effects of octreotide were then evaluated using open field and elevated plus-maze tests. Following that, mice were intraperitoneally given a single convulsive dosage of PTZ (60 mg/kg) and then monitored for 30 min for symptoms of seizures. Finally, the antioxidant capacity of brain tissue and histopathological changes in the hippocampus were investigated. Octreotide therapy for seven days at 50 or 100 µg/kg was more effective than diazepam in preventing acute PTZ-induced seizures (P < 0.05). Furthermore, both octreotide dosages revealed substantial anxiolytic effects in open-field and elevated plus-maze tests compared to untreated rats. Nonetheless, octreotide's anxiolytic impact was less effective than diazepam's. On the other hand, octreotide also suppressed neuronal apoptosis and attenuated oxidative stress. Our results suggest that chronic administration of octreotide has anticonvulsant, anxiolytic, and antioxidant activity in the male Wistar rat model.

Effect of dietary inclusion of atorvastatin, garlic, and dill on growth performance, antioxidant defense, gut, and cardio-pulmonary function, and lipogenesis in broiler chickens.

In the present study, we evaluated the antihyperlipidemic and antioxidant effects of garlic and dill in comparison with atorvastatin to combat lipogenesis in broiler chickens. A total of 400 1-day-old chicks (Ross 308 strain) were randomly distributed into four experimental diets. Dietary treatments included a control diet, the control diet plus atorvastatin at 20 mg/kg, the control diet plus garlic dry powder (GDP) at 7.5 g/kg, and the control diet plus dill dry powder (DDP) at 7.5 g/kg. Chicks were maintained on experimental diets for 42 days under the recommended environmental conditions set out by the strain management manual. The results showed that weight gain, feed conversion ratio (FCR), and duodenal, jejunal, and ileal dimensions of villi (height, width, and the surface absorptive area) were improved by in-feed atorvastatin, GDP, or DDP when compared to the control (P < 0.05). The inclusion of atorvastatin or phytobiotic products increased circulatory levels of nitric oxide (NO) but decreased circulatory levels of malondialdehyde (MDA), triacylglycerol (TAG), and low-density lipoproteins cholesterol (LDL), with concomitant reductions in the T, R, and S waves amplitudes in the Lead 2 electrocardiogram (ECG) (P < 0.05). Dietary supplements caused an up-regulation of inducible nitric oxide synthase (iNOS), superoxide dismutase 1 (SOD1), and glutathione peroxidase (GPX) but reduced the expression of key hepatic lipogenic enzymes (fatty acid synthase (FAS) and hydroxy-methylglutaryl-CoA reductase (HMGCR) (P < 0.05). In conclusion, feed supplementation with atorvastatin, GDP, or DDP suppressed lipogenesis, enhanced antioxidant response, and improved gut and cardio-pulmonary function in broiler chicks subjected to hypobaric hypoxia.

Effect of sage (Salvia officinalis L.) extract in antioxidant status and intestinal morphology of pulmonary hypertensive chickens.

OBJECTIVES: The effects of dietary sage on the growth performance, antioxidant status, intestinal mucosa morphology, and pulmonary hypertensive response were investigated in broiler chickens with pulmonary hypertension. METHODS: Chicks (Ross 308) were reared under cold stress for 35 days and treated with 0.05% vitamin C (positive control) and 0 (control), 0.1 or 0.2% sage extracts, then performance, oxidant and antioxidant status, and intestinal morphology were evaluated. RESULTS: The index of pulmonary hypertension (RV:TV) was decreased, and weight gain (days 22-35) was increased in all treatments (except for sage 0.1%) compared with control (P < 0.05). Lipid peroxidation was decreased, whereas the activity of antioxidant enzymes (GPX, CAT, and SOD) was increased in the sage 0.2% group compared with control (P < 0.05). In the lung, SOD, CAT, and GPX transcripts were decreased in the sage 0.2% group compared with control (P < 0.05). In the right ventricle of the heart, SOD and CAT transcripts were increased in the sage 0.2% group compared with other groups of chickens, whereas GPX transcript was decreased (P < 0.05). The jejunal villus length in the chickens fed sage was significantly lower than in control (P < 0.05). The ileal villus width, villus surface area, and lamina proporia thickness in the chickens fed sage (0.2%) were increased compared with control (P < 0.05). CONCLUSIONS: Dietary supplementation of sage (0.2%) could modulate pulmonary hypertensive response, improve antioxidant status (enzymatic activity), intestinal morphometry, and absorptive surface in the broiler chickens.

Increased serum lipofuscin associated with leukocyte telomere shortening in veterans: a possible role for sulfur mustard exposure in delayed-onset accelerated cellular senescence.

BACKGROUND: Sulfur mustard (SM) is a toxic gas that causes chronic inflammation and oxidative stress leading to cell senescence. This study aimed to evaluate two indicators of biological aging (i.e., serum lipofuscin level and leukocyte telomere length) and assess their relationship based on the severity of SM exposure in the long term. METHODS: The study was performed on two groups of male participants. 1) SM-exposed group (exposed to SM once in 1987), 73 volunteers. 2) Non-exposed group, 16 healthy volunteers. The SM-exposed group was categorized into three subgroups based on the severity of SM exposure and body damage (asymptom, mild, and severe). The blood sample was prepared from members of each group. The serum lipofuscin, TGF-ß, malondialdehyde (MDA), c-reactive protein (CRP), and leukocyte telomere length (TL) were measured in all participants. RESULTS: The MDA level was increased in the SM-exposed group (mean = 39.6 µM, SD = 16.5) compared to the non-exposed group (mean = 21.1 µM, SD = 10.3) (P < 0.05). The CRP level was also increased in the SM-exposed group (mean = 5.12 mg/l, SD = 3.36) compared to the non-exposed group (mean = 3.51 mg/l, SD = 1.21), while the TGF-ß level was decreased (P < 0.05) in the SM-exposed group (mean = 52.6 pg/ml, SD = 18.7) compared to the non-exposed group (mean = 68.9 pg/ml, SD = 13.8). The relative TL was shorter in the SM-exposed group (mean = 0.40, SD = 0.28) than in the non-exposed group (mean = 2.25, SD = 1.41) (P < 0.05). The lipofuscin level was higher in the total SM-exposed group (mean = 1.44 ng/ml, SD = 0.685) than in the non-exposed group (mean = 0.88 ng/ml, SD = 0.449) (P < 0.05). The MDA and CRP levels were increased in the SM-exposed subgroups of asymptom, mild, and severe than the non-exposed group, while TGF-ß level and TL were decreased in those subgroups. The lipofuscin level was higher in the SM-exposed subgroups of mild and severe than in the non-exposed group. The regression analysis determined a negative correlation between lipofuscin level and TL. The lipofuscin/TL ratio was higher in the total SM-exposed group (mean = 6.36, SD = 5.342) than in the non-exposed group (mean = 0.51, SD=0.389). This ratio was also higher in the SM-exposed subgroups of asymptom, mild, and severe than in the non-exposed group. The lipofuscin/TL ratio did not differ between mild and severe subgroups. CONCLUSION: The delayed toxicity of SM is associated with chronic oxidative stress, continuous inflammatory stimulation, increased lipofuscin, and telomere shortening. Future studies are needed to verify the suitability of serum lipofuscin to telomere length ratio in determining the severity of SM toxicity.

Co-culturing or conditioned medium of Sertoli cells: Which one supports in vitro maturation of bovine oocytes and developmental competency of resulting embryos?

BACKGROUND: Sertoli cells (SCs) as supportive cells in the seminiferous tubule play an essential role in the nutrition and development of adjacent cells by secreting several beneficial growth factors, stimulators and cytokines which can be conceived to improve the developmental competency of oocyte or embryo in the co-culture system. OBJECTIVES: This study aimed to improve the maturation of bovine oocytes and consequently the development of resulting embryos in co-culture with SCs and their conditioned medium (CM). METHODS: The retrieved cumulus-oocyte complexes (COCs) from the abattoir-derived ovaries were matured in maturation medium alone (control group), in co-culture with ovine SCs (co-culture group), and in presence of 10% CM prepared in 33°C and 39°C (CM33 and CM39 groups). The nuclear maturation competency and subsequent embryo development rate of cultured COCs in all groups were evaluated. RESULTS: The results of this study showed that SCs and CM increased meiosis resumption from GV to the MII compared to the control group, significantly (p < 0.05). Besides, the degenerated oocytes in the co-culture group were significantly higher than those in the control, CM33 and CM39 groups (p < 0.05), and the lowest cleavage rate belonged to the co-culture group (p < 0.05). The blastocyst rate was also lower in the co-culture group than other groups and there was a significant difference between the control and two CM groups (p < 0.05). CONCLUSIONS: The Sertoli cells can be suitable for co-culturing with oocytes during IVM but detrimental for subsequent embryo development. In turn, Sertoli cell-derived conditioned medium (SC-CM) can provide sufficient bioactive materials for COCs to enhancing oocyte competence and embryo development.

Concomitant use of relative telomere length, biological health score and physical/social statuses in the biological aging evaluation of mustard-chemical veterans.

Sulfur mustard (SM) is a toxic gas that has been used as a chemical weapon in wars. After many years, SM-exposed people are still suffering from its side effects such as biological and premature aging. This study was aimed to evaluate biological aging rate via involving biological health scoring (BHS), relative telomere length (TL) and different physical/social variables i.e. marital and smoking statuses, body mass index, salary and educational levels. BHS was calculated according to measurement of 18 biomarkers related to function of four physiological systems (endocrine, inflammatory, cardiovascular and metabolic systems) and two organs (liver and kidney). The volunteers were 442 individuals exposed to SM gas in 1987 and 119 healthy individuals as non-exposed group. Each group was divided based on leukocyte relative TL (short, intermediate and long). Our data showed an inverse correlation between BHS and relative TL in two groups. The BHS was significantly higher in SM-exposed group than non-exposed group, especially in the participants with short and intermediate TL. The BHS had also a positive correlation with smoking and BMI parameters, and a negative correlation with salary and educational levels in the participants with shorter telomeres; and SM strengthened these correlations in the shorter telomeres. It is concluded that the higher BHS along with shorter relative TL that are indices for lower health quality and biological aging, could be used in the health evaluation of non- and SM-exposed people; and involving of BHS, TL and physical/social covariates could be useful to make this evaluation more accurate.

Sulfur mustard and biological ageing: A multisystem biological health score approach as an extension of the allostatic load in Sardasht chemical veterans.

BACKGROUND: Mustard gas as a chemical weapon has been used in wars and its long-term side effects are substantial in the human body. This study was aimed to apply an extension of allostatic load, known as synthetic biological health score (BHS), including the wear-and-tear of four physiological systems (endocrine, inflammatory, cardiovascular and metabolic systems) and two organs (liver and kidney) to estimate biological aging caused by sulfur mustard (SM) gas poisoning. METHODS: The plasma samples were prepared from two following groups of people; 1) 446 individuals exposed to SM gas in 1987. 2) 115 healthy members, at the same range of age and residence, as the non-exposed group. These people were chosen from the same patients' families and have not had any exposure to SM or pulmonary dysfunction. To estimate BHS, 18 blood-derived biomarkers of the mentioned systems and organs were measured and the relative contribution of many social and body parameters across the age groups was explored. RESULTS: It was revealed, in BHS calculation, the cardiovascular system had the most effect. Also, in the SM group, BHS was significantly higher than in the control group. This feature has a positive correlation with physical parameter (BMI) and a negative correlation with social parameters (salary and educational levels). CONCLUSION: The multisystem BHS could be useful in the evaluation of biological aging due to SM exposure. Social (education, good-paying job) and physical (BMI) parameters could influence BHS and the higher BHS indicates the progress of biological aging due to mustard gas.

Pomegranate peel as a phytogenic in broiler chickens: Influence upon antioxidant, lipogenesis and hypotensive response.

The aim of this study was to evaluate antioxidant, antihyperlipidemic and hypotensive properties of pomegranate peel (PP) on antioxidant status, fat deposition, lipid peroxidation and pulmonary hypertensive response in broiler chickens. A total of 375 one-day-old male broilers (Cobb 500) were randomly assigned to five treatments included dietary PP levels of 0, 2.5, 5.0, 7.5 and 10 g/kg. Supplementation of PP at 7.5 and 10 g/kg resulted in significant upregulation of hepatic catalase (p < 0.004) and superoxide dismutase1 (SOD1; p < 0.05), which reflected in decreased concentration of circulatory malondialdehyde (MDA). Dietary inclusion of PP at 7.5 and 1.0 g/kg significantly decreased serum concentrations of triglycerides (p < 0.004) and cholesterol (p < 0.006) with concomitant decrease in abdominal fat deposition (p < 0.05). The antihyperlipidemic effect of PP was mediated through down-regulation of peroxisome proliferator activated receptor alpha (PPARα). Hypotensive effect of PP was also observed at 7.5 and 10 g/kg as reduced heart weight and the right-to-total ventricular weight ratio (RV/TV) and decreased mortality from pulmonary hypertension. The hypotensive property of PP was associated with increased concentration of serum nitric oxide. In conclusion, this study revealed antioxidative, antihyperlipidemic and hypotensive effects of PP at 7.5 and 10 g/kg in broiler chickens exposed to hypobaric hypoxia. Health-beneficial effects of PP suggest this product as a promising multi-functional phytogenic feed additive for broiler chickens.

Macular hole and submacular hemorrhage secondary to retinal arterial macroaneurysm - successfully treated with a novel surgical technique.

Purpose: To present a 65-year-old patient with macular hole (MH) and submacular hemorrhage (SMH) secondary to a ruptured retinal arterial macroaneurysm (RAM) which was successfully treated with a novel surgical technique. Patient: A 65-year-old woman presented with a 1-week history of sudden-onset visual loss in her right eye. Her best-corrected visual acuity (BCVA) was hand motion in the right eye. Her fundus examination revealed intraretinal and massive subretinal hemorrhage with macular involvement. An MH with an approximate diameter of 600 microns was also noted. The patient was treated by a standard pars plana vitrectomy (PPV) and internal limiting membrane (ILM) peeling, followed by 10 µg recombinant tissue plasminogen activator injection into the submacular space via the MH with a 25-gauge vitrectomy probe. BCVA of the patient increased to 20/320 and 20/60, one and four weeks after surgery, respectively. Optical coherence tomography (OCT) images confirmed the complete anatomical closure of the MH and the restoration of the outer retinal layers such as the external limiting membrane and the ellipsoid zone. Conclusion: This case report expands our knowledge about the management of MH in combination with SMH after a ruptured RAM. We suggest the use of a vitrectomy probe and MH for subretinal recombinant tissue plasminogen activator (rtPA) injection.

Impacts of in vitro thermal stress on ovine epididymal spermatozoa and the protective effect of ß-mercaptoethanol as an antioxidant.

Most aspects of reproductive function including spermatogenesis, oocyte growth and maturation, early embryonic development, fetal and placental growth, and lactation can be affected by thermal stress. Furthermore, it has been shown that oxidative stress involves in the pathology of thermal stress. Therefore, the aim of this study was to investigate the impacts of thermal stress on the ovine mature epididymal spermatozoa extracted from testes of slaughtered rams in the presence or absence of an antioxidant. Epididymal spermatozoa were incubated at scrotal (32.00 ˚C), normal body (39.00 ˚C), and hyperthermic temperatures (41.00 ˚C) for 4 hr in the presence or absence of 1 mmol L-1 ß-mercaptoethanol. The results demonstrated the high sensitivity of ram epididymal spermatozoa to the hyperthermic temperature at in vitro conditions. In comparison with scrotal temperature, quality parameters of spermatozoa were negatively affected by increase in temperature, as such in the spermatozoa incubated at hyperthermic temperature significant decrease was observed in the viability, DNA integrity and in the majority of motility parameters. Moreover, concentration of lipid peroxidation by-products, thiobarbituric acid reactive substances, were significantly increased. The findings showed that using antioxidant during incubation period had significant protective effect on the viability and motility of incubated spermatozoa not only at the hyperthermic temperature, but also at the scrotal and normal body temperatures. In conclusion the ovine epididymal spermatozoa were sensitive to in vitro thermal stress and it seems that this sensitivity was partly related to the oxidative stress.

Effect of Shirazi thyme on oxidant status and absorptive surface area of the intestine in cold-induced pulmonary hypertensive broiler chickens.

The effects of Shirazi thyme as a medicinal plant on oxidant status (lipid peroxidation, protein oxidation, total antioxidant capacity, and catalase activity) and absorptive surface area were measured in three segments of the small intestine in cold-induced pulmonary hypertensive chickens. Birds were reared at four groups (thyme 0, 0.25, 0.50, and 1.00 % of diet) for 42 days. To induce pulmonary hypertension, the temperature was gradually decreased. The body weight was increased in thyme-0.25% birds compared to control ones, while it was decreased in thyme-1% of birds. The feed consumption was only increased in thyme-1.00% birds. The feed conversion ratio was lower in thyme-0.25% birds and higher in thyme-1.00% birds than control ones. The duodenal and jejunal villus surface area was lower in thyme-1.00% birds than control ones, while it was greater in the thyme-0.50% birds. The ileal villus surface area and duodenal lamina propria thickness were also greater in thyme-0.50% birds. Lipid peroxidation was only decreased in the duodenum and ileum of thyme-0.50% birds compared to control ones, whereas it was increased in the duodenum and jejunum of thyme-1.00% birds. Catalase activity was only elevated in the duodenum and jejunum of thyme-1.00% fed chickens. Total antioxidant capacity was increased in the duodenum, jejunum, and ileum of thyme-0.50% birds. It is concluded that the Shirazi thyme has beneficial effects on growth performance, intestinal absorptive surface area / secretory system, and pulmonary hypertension response at low doses (0.25 and 0.50% of diet), whereas high dose (1.00% of diet) of this plant may be toxic.

Observation of nine previously reported and 10 non-reported SLC4A11 mutations among 20 Iranian CHED probands and identification of an MPDZ mutation as possible cause of CHED and FECD in one family.

BACKGROUND/AIMS: SLC4A11 is the only known causative gene of congenital hereditary endothelial dystrophy (CHED). Mutation screenings have shown that most but not all patients with CHED harbour mutations in SLC4A11, suggesting that other CHED-causing genes may exist. We aimed to screen SLC4A11 in Iranian patients to learn the mutation spectrum of this gene among Iranians and to gain further knowledge on potential contribution of other genes to CHED aetiology. METHODS: SLC4A11 was screened in 21 Iranian patients with CHED by sequencing. Previously unreported variations were checked in at least 200 controls, and segregation analysis within families and bioinformatics predictions on effects of variations were performed. Exome sequencing was done for the single patient without an SLC4A11 mutation and for her parents. RESULTS: Nine previously reported and 10 unreported SLC4A11 mutations were observed among 20 patients; a mutation was not found in one patient. A mutation in MPDZ was identified as the only candidate cause of CHED in this patient. Her mother who carried the same mutation was diagnosed with Fuchs endothelial corneal dystrophy (FECD). CONCLUSION: SLC4A11 mutations are the usual cause of CHED in Iranians. The 10 novel mutations observed contribute significantly to the approximately 85 mutations reported since discovery of the role of the gene in CHED pathogenesis more than 10 years ago. MPDZ mutations may be a cause of CHED and even FECD in a minority of patients. Proposed functions of MPDZ with respect to tight junctions and maintenance of the corneal endothelial barrier are in accordance with a role in corneal endothelial pathobiology.

Identification of reliable reference genes for quantitative real-time PCR in ovary and uterus of laying hens under heat stress.

The main stage in real-time quantitative PCR is a quantification of gene transcriptomes, in which suitable use of reliable reference genes is critical to normalize accurately. To determine the most stable reference genes in laying hens under heat stress, from a panel of nine typical candidate reference genes, the mRNA transcript of ACTB, HMBS, HPRT1, RPL13, RPL32, 18SrRNA, TBP, TFRC, and YWHAZ was evaluated in the ovary and uterus of both control and heat-stress groups of laying hens. Forty 23-week-old White Leghorn laying hens were housed in two rooms. The control (n = 20) and heat-stress (n = 20) groups were maintained at 21-23 °C and 36-38 °C for 8 weeks respectively. Analysis of this set of genes was done with BestKeeper, geNorm, and NormFinder software programs to find the most stable ones. Candidate reference genes ranked in the uterus of heat-stress and control groups of hens included YWHAZ, HPRT1, HMBS, RPL13, TFRC, ACTB, TBP, RPL32, and 18SrRNA; those in the ovary were YWHAZ, HPRT1, TFRC, HMBS, RPL13, TBP, RPL32, ACTB, and 18SrRNA. The overall results indicated that the most stable genes are YWHAZ, HPRT1, HMBS, RPL13, TFRC, TBP, ACTB, RPL32, and 18SrRNA respectively. In addition, the combination of YWHAZ, HPRT1, and HMBS is suggested as the most stable reference group of genes for more accurate quantitative data normalization in the ovarian and uterine tissues of laying hens under control and heat stress conditions. Lay summary Heat stress influences the expression of many genes in the reproductive tissues of birds. Accurate evaluation of these changes via real-time quantitative PCR depends on the determination of reliable reference genes. In this study, nine candidate housekeeping genes were evaluated, and the most stable were YWHAZ, HPRT1, HMBS, RPL13, TFRC, TBP, ACTB, RPL32, and 18SrRNA.

ELECTROCARDIOGRAM ANALYSIS OF THE GOLDEN ( CHRYSOLOPHUS PICTUS) AND SILVER ( LOPHURA NYCTHEMERA) PHEASANTS.

The aim of this study was to describe normal electrocardiogram patterns and values in two species of conscious pheasants (golden and silver). The standard bipolar and augmented unipolar limb leads electrocardiograms were recorded in the birds. The wave forms were analyzed in all leads at 50 mm/sec and at 10 mm = 1 mV to determine PR, QRS, ST, and QT durations and P, net QRS complex, and T amplitudes. The polarity of each wave form was tabulated in all leads. The mean electrical axis for the frontal plane, calculated using leads II and III, was mostly negative in two species (-43 ± 28.0° in golden and -92 ± 10.0° in silver pheasant). The P wave was predominantly positive in most of the leads. The pattern of wave forms of the QRS complexes were different (Rs, QS, rS, R, RS, QR, qR). The T wave was almost positive in leads I, II, III, and aVF. The heart rates of the birds were mean (± SD) of 320 ± 36 beats/min (golden) and 314 ± 31 beats/min (silver). Description of normal electrocardiogram parameters will facilitate a better understanding of electrocardiogram changes of unhealthy birds.

Evaluation of hepatic lipogenesis and antioxidant status of broiler chickens fed mountain celery.

BACKGROUND: Fatness is an unwanted side effect of genetic selection in broiler chickens. In this study, we introduce mountain celery powder as a feed supplement to suppress lipogenesis and improve antioxidant status in broiler chickens. Male broiler chicks (Ross 308) were fed a control diet or a diet that includes mountain celery (MC) at 7.5 gkg-1over 42 days. RESULTS: Body weight gain and feed conversion ratio significantly (P < 0.05) improved in chicks fed MC. A highly significant down-regulation of genes involved in hepatic lipogenesis including acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), malic enzyme (ME), and lipoprotein lipase (LPL) was observed in the liver of chickens fed MC. These birds, however, had greater compensatory upregulation in antioxidative genes SOD1 and catalase in the liver compared to the birds that received the control diet. Birds received MC had significantly lower level of lipid peroxidation (1.59 µmol/L serum malondialdehyde) compared to birds from the control group (3.57 µmol/L; P = 0.0024). Birds fed MC had significantly (P < 0.05) lower circulatory concentrations of triacylglycerols, cholesterol, and LDL but higher concentrations of HDL. Relative liver weight and abdominal fat deposition were significantly reduced by feeding MC. CONCLUSIONS: It can be concluded that feeding birds MC significantly suppresses hepatic lipogenesis by down-regulating key hepatic lipogenic enzyme genes and boosts antioxidant capacity by up-regulating hepatic antioxidantive genes.

In vitro study of matrix metalloproteinases 1, 2, 9, 13 and serum amyloid A mRNAs expression in equine fibroblast-like synoviocytes treated with doxycycline.

Application of synthetic matrix metalloproteinases (MMPs) inhibitors, such as doxycycline is one of the possible therapeutic options for osteoarthritis. However, little is known about the protective mechanism of doxycycline in equine models on MMPs inhibitors as well as on serum amyloid A (SAA) gene expression. This study investigated the effects of doxycycline on mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA of equine fibroblast-like synoviocytes (FLSs). The FLSs were established from synovial fluids of clinically normal metacarpophalangeal joints of 6 skeletally mature horses. The cells were treated with either 10 or 100 µg/mL of doxycycline for 48 h. The mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA were assessed using real-time polymerase chain reaction (PCR). Treatment with doxycycline resulted in significantly decreased mRNA expression of MMP-1 in FLSs at both concentrations (P = 0.001). No significant differences were detected among groups for MMP-2, MMP-9, and MMP-13 (P > 0.05). Only a tendency towards a decrease in mRNA expression level of SAA in the presence of doxycycline could be detected. Doxycycline inhibits MMP-1 gene expression at the transcript level. These findings indicate that doxycycline can protect the articular environment through inhibition of MMP-1 at transcript level.

L'application d'inhibiteurs synthétiques des métalloprotéinases de la matrice (MMP), telle que la doxycycline, est une des options thérapeutiques possibles pour l'ostéoarthrite. Toutefois, peu de choses sont connues sur le mécanisme protecteur de la doxycycline dans les modèles équins des inhibiteurs des MMP, de même que sur l'expression génique de l'amyloïde sérique A (SAA). La présente étude visait à déterminer les effets de la doxycycline sur l'expression de l'ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA des synoviocytes équins apparentés aux fibroblastes (FLS). Les FLS ont été établis à partir du liquide synovial provenant d'articulations métacarpo-phalangiennes cliniquement normales de six chevaux squelettiquement matures. Les cellules ont été traitées avec 10 ou 100 µg/mL de doxycycline pendant 48 h. L'expression d'ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA a été évaluée par réaction d'amplification en chaîne par la polymérase en temps réel. Le traitement avec la doxycycline a causé une diminution significative de l'expression de MMP-1 par les FLS et ce pour les deux concentrations (P = 0,001). Aucune différence significative ne fut détectée parmi les groupes MMP-2, MMP-9, et MMP-13 (P > 0,05). Seulement une tendance à la diminution de l'expression d'ARNm de SAA en présence de doxycycline pouvait être notée. La doxycycline inhibe l'expression génique de MMP-1 à l'étape de la transcription. Ces informations indiquent que la doxycycline peut protéger l'environnement articulaire en inhibant MMP-1 à l'étape de la transcription.(Traduit par Docteur Serge Messier).

Effect of heat stress on the gene expression of ion transporters/channels in the uterus of laying hens during eggshell formation.

Heat stress is a problem in laying hens as it decreases egg quality by decreasing eggshell mineralization. Heat stress alters gene expression, hence our aim was to investigate effects of heat stress on gene expression of ion transport elements involving in uterine mineralization (TRPV6, CALB1, ITPR3, SCNN1G, SLC4A4, KCNJ15, SLC4A9, and CLCN2) by real time quantitative PCR. Forty 23-week-old White Leghorn laying hens were housed in two rooms. The control group (n = 20) was maintained at 21-23 °C, and the heat stress group (n = 20) was exposed to 36-38 °C for 8 weeks. All parameters of egg quality including egg weight, surface area, volume, and eggshell weight, thickness, ash weight, and calcium content were decreased in the heat stress group compared to the control group (by 26.9%, 32.7%, 44.1%, 38.4%, 31.7%, 39.4%, and 11.1%, respectively). Total plasma calcium was decreased by 13.4%. Levels of ITPR3, SLC4A4, and SLC4A9 transcripts in the uterine lining were decreased in the heat stress group compared to the control group (by 61.4%, 66.1%, and 66.1%, respectively). CALB1 transcript level was increased (by 34.2 fold) in the heat stress group of hens compared to controls. TRPV6, SCNN1G, KCNJ15, and CLCN2 transcript levels did not significantly differ between control and heat stress groups of laying hens. It is concluded that the down-expression of ITPR3, SLC4A4, and SLC4A9 genes may impair transportation of Cl-, HCO3-, and Na+ in eggshell mineralization during heat stress. Increased CALB1 gene expression may increase resistance of uterine cells to detrimental effects of heat stress.