Spermatogenesis regeneration by transfected spermatogonial stem cells in infertile roosters through testicular transplantation.

Investigations pertaining to spermatogonial stem cells (SSCs) have led to the use of these cells in a variety of fields including infertility treatments, production of transgenic animals, and genome editing. The aim of the present study was to investigate the plausibility of regenerating spermatogenesis in infertile roosters by transplanting transfected SSCs into testes. Spermatogonial stem cells were isolated and cultured for seven days. Afterward, pDB2, a plasmid vector carrying a reporter gene, GFP, was transfected into the SSCs. Transfected SSCs were transplanted into the left testis of infertile roosters. Tissue samples from the recipients' testes were obtained six weeks after the transplantation and transplanted SSCs were observed in the basement membrane. After eight weeks, GFP-positive spermatozoa were observed in collected semen from the recipient roosters and GFP gene in spermatozoa was confirmed using PCR. The recipient roosters were mated with hens. Hatchlings were visually checked and their tissue samples were tested by PCR to identify transgenesis but both of them were negative. Overall, it seems that regeneration of spermatogenesis in roosters via transfected SSCs is possible but more studies are need to produce recombinant proteins by this way.

Applying mindfulness training to enhance the mental toughness and emotional intelligence of amateur basketball players.

OBJECTIVES: Mindfulness is a proven treatment in clinical settings. Our objective was to examine the feasibility and effect of a mindfulness-training program on the mental resilience and emotional intelligence of amateur basketball players. METHODS: This was a parallel-group, pre- and post- test, randomised controlled trial. Thirty male amateur male basketball players from Tehran, Iran, were assigned randomly into experimental and control groups (n = 15 each). Outcomes were measured using the mindfulness sport inventory, mental toughness (MT) questionnaire and emotional intelligence questionnaire. Data were analysed using ANCOVA and MANCOVA. RESULTS: Mindfulness scores were significantly greater in the intervention group than in the controls following training. The intervention also increased overall MT and all of the component sub scores. The same applied to emotional intelligence and all of its components. CONCLUSIONS: These findings may have implications on sport mindfulness training in increasing the MT and emotional intelligence of athletes.

Developing a puncture-free in ovo chicken transfection strategy based on bypassing albumen nucleases.

Chicken is a dual-purpose animal important from both agricultural and medical aspects. Even though significant improvements have been made in chicken transgenesis technologies, chicken genome manipulation has not been widely used in developmental biology. This study was aimed to evaluate chicken egg white nuclease properties and thereof plausibility of devising an in vivo transfection technology without causing physical damage to the embryo. First, the nuclease activity of egg albumen was assessed. The egg white nucleases were strongly active in degrading DNA and RNA. The egg white DNase activity was comparable to commercially available DNase-I. Nuclease activities were also assessed after heating, proteinase K, or EDTA treatment. Unlike proteinase K, both heating and EDTA were noticeably effective for the nuclease inactivation. Simultaneous application of lipoplex form of DNA (1 µg pDB2: 3 µl Lipofectamine2000) and EDTA showed a synergistic effect in protection against egg white nucleases. Finally, we injected the lipoplexes with or without EDTA close to the embryo at day0, but outside the embryonic epiblast. Implementation of a scrutinized PCR assay indicated that transfection took place only when EDTA was complemented to the lipoplexes. The transfection rate of day4 embryos and the hatched chicks were 54.5 and 30.0%, respectively. EGFP expression was detected in two out of three transgenic chicks. In conclusion, this study provided a detail analysis of chicken egg albumen nuclease properties and suggested the feasibility of developing a puncture-free handmade technology for transfection of the chicken embryo.

A SNP in the 3'-untranslated region of AMPKγ1 may associate with serum ketone body and milk production of Holstein dairy cows.

AMPK is the key switch for providing the energy balance between cellular anabolic and catabolic processes. In this study, we aimed to screen the PRKAG1 (AMPKγ1) gene in high, moderate, and low producing Holstein dairy cows. A sample of 100 pregnant dairy cows, comprising 41 high, 33 moderate, and 26 low milk yields were selected from three large dairy herds in Isfahan province of Iran. Body condition score (BCS) was estimated before parturition while beta hydroxyl butyric acid (BHBA) as a measure of ketone bodies was measured at the fifth day postpartum. In addition, using three primer pairs covering exons 2-11 and 3'-UTR of the PRKAG1 gene, a random sample of 10 high milk yield dairy cows were amplified and sequenced. The sequencing results showed the presence of a T12571C mutation in intron 6 and a T14280C mutation in the 3'-untranslated region (UTR) of the PRKAG1 gene. Following a PCR reaction for amplification of the 3'-UTR amplicons, single strand conformation polymorphism (SSCP) assay was implemented for discrimination of the mutation in the studied population. Then, we evaluated if the mutation associates with the BCS, serum BHBA level, and production traits. The experimental analysis showed that the mutated allele significantly increased the BHBA level, BCS, as well as milk and protein yield. Bioinformatic study revealed that this 3'-UTR mutation distorts the target site of mir-423-5p microRNA which is one of the most highly expressed microRNAs in the bovine mammary gland, liver, and kidney. Given the role of AMPK in energy metabolism, the newly identified 3'-UTR mutation highlights the importance of AMPK and suggests a role of miRNAs for regulation of cellular metabolism, metabolism disorders, and production traits in Holstein dairy cows.

Stabilization of heavy metals in wastewater treatment sludge from power plants air heater washing.

In the present investigation, for the first time in Iran, dewatered sludge waste from the air heater washing wastewater treatment of a thermal power plant was subject to investigation with regard to cement-based stabilization in order to reduce the mobility of heavy metals. Studies of the chemical composition, mineralogy, morphology and leaching characteristics of the untreated and treated waste were conducted. The stabilization was done with two cement-based mix designs with different waste/cement ratios in different samples. In the cement stabilization process the leaching of V, Ni, Zn and Cr could be decreased from 314.1, 209.1, 24.8 and 5.5 mg L(-1), respectively, in the untreated waste to 6.02, 32.11, 3.57 and 3.68 mg L(-1) in a mixture with 20% waste/cement ratio, while during stabilization with cement + sand, the toxicity characteristic leaching procedure (TCLP) leachate contents of the mentioned metals were decreased to 4.24, 16.2, 2.51 and 2.2 mg L(-1) for the same waste/cement ratio. Stabilization with sand/cement seemed to be more efficient in reducing metal leaching from the waste. X-ray diffraction studies showed that portlandite, calcite, lime, larnite and quartz were produced during the stabilization process.