Assessing the Impact of the Novel Sperm Selection Technique 'Annexin-V Coated Polystyrene Bead Technique' on Mouse Assisted Reproductive Techniques Outcomes: Preliminary Findings.

ICSI is one of the most commonly used techniques to treat infertility. The sperm selection for the procedure is done 'randomly' by the embryologist according to the motility and morphology parameters which is known not to reflect the potential of a sperm for fertilization, pregnancy and a healthy childbearing. Since the apoptosis rate is higher in sperm cells of infertile patients, it is more likely to choose an apoptotic sperm by the 'random selection method'. We recently introduced a novel sperm selection technique namely 'Annexin-V coated polystrene bead technique'(APB-Tech), for the selection of non-apoptotic sperm cells. The principal of the technique is based on the binding affinity of an apoptotic sperm to 'Annexin-V covered beads' enabling to distinguish a viable and a healthy sperm by light microscopy. The aim of the present study was to observe the effects of this technique on ICSI outcomes in mice. Sibling-oocyte trial was conducted and the outcome measures were compared with the results of traditional sperm selection method. Embryo and blastocyst qualities and blastocyst development rates were significantly increased in APB-Tech group, while the other parameters were not affected. Promising results obtained from the technique reflect its promising potential as a new and powerful tool for sperm selection and thus infertility techniques.

Sperm selection with Annexin-V coated polystrene bead technique (APB-Tech): A novel and reliable method for the microscopic selection of viable and non-apoptotic sperm to be used for intracytoplasmic sperm injection.

Infertility is a growing health problem that affects 20% of couples who want to conceive. 'Intracytoplasmic sperm injection' (ICSI) is a commonly used assisted reproduction treatment technique that offers the couples to overcome a wide variety of infertility reasons, including female, male and unexplained infertility. A crucial step in ICSI is choosing the correct sperm with the highest potential to form a healthy embryo and thus a healthy offspring. The sperm selection strategies aim to obtain a sperm population with better motility and morphology, which are insufficient to predict a sperm's reproductive potential. The elimination of apoptotic sperms, which is shown to be higher in infertile males and which is shown to affect embryo development and reproductive cohort significantly should also be included in the selection strategies. Among all sperm selection methods, there are only a few which can eliminate apoptotic sperm, but because they need extra types of equipments, a long training period, and high costs, they couldn't find place in the most commonly used techniques in an IVF lab. Selecting the non-apoptotic sperm cells will help us choose a sperm that is more likely to be chosen by the natural selection mechanisms and thus will help to mimic the natural conception more. The study aimed to develop a novel, easy and a harmless individual sperm selection technique to enable choosing non-apoptotic viable sperm cells via light microscopy without any need for extra equipment, education and cost to be used for ''ICSI'. The technique is based on the binding ability of Annexin-V covered polystrene beads to the externalized phosphatidylserine at the outer leaflet of an apoptotic sperm's plasma membrane. After Annexin-V covered polystyrene beads were prepared, beads obtained were attached to BALB-c mice sperm, and the technique is optimized to obtain the most efficient attachment conditions. The results are then compared with the results of four well-known reliable apoptosis detection techniques to test the validity and sperm survival test to test the toxicity of the technique. The method is proven to be '''reliable' by comparing it with the results of well-known techniques, including TUNEL and SCSA (sperm chromatin structure assay), and '''safe' by showing its non-toxicity via sperm survival test (SST). In addition, the method enables the selection of sperm cells more closer to naturally-chosen ones from a pool of sperm that should not be allowed to be randomly chosen during microinjection. To date, it was impossible to distinguish a non-apoptotic sperm without harming it or without needing additional equipment other than a routine IVF lab and extra training other than routine andrology work. The technique is named as 'Annexin-V coated polystyrene bead technique (APB-Tech)'. Based on our results, further studies on APB-Tech should be focused on the possible improvement of ICSI outcomes and, thus, success rates.

Performance of capecitabine in novel combination therapies in colorectal cancer.

Colorectal cancer is one of the most common cancers throughout the world, and no definitive cure has ever been found. Perhaps a new insight into the effectiveness of chemotherapy drugs could help better treat patients. Targeted therapies have significantly improved the median overall survival of colorectal cancer patients. One of the standard chemotherapy regimens used for colorectal cancer is capecitabine, which is important in monotherapy and combination therapies. Capecitabine, with other chemotherapeutic agents (irinotecan, oxaliplatin, perifosine, 17-allylamino-17-demethoxygeldanamycin, aspirin, celecoxib, statins, quinacrine, inositol hexaphosphate and inositol, cystine/theanine, curcumin, and isorhamnetin), and biological ones (antibodies) plays an important role in the inhibition of some signaling pathways, increasing survival, reducing tumor growth and side effects of capecitabine. However, some drugs, such as proton pump inhibitors, are negatively related to capecitabine; therefore, the purpose of this work is to review and discuss the performance of capecitabine combination therapies in colorectal cancer.

Overexpression of polyphosphate kinase gene (ppk) increases bioinsecticide production by Bacillus thuringiensis.

Polyphosphate (polyP), synthesized by polyP kinase (PPK) using the terminal phosphate of ATP as substrate, performs important functions in every living cell. The present work reports on the relationship between polyP metabolism and bioinsecticide production in Bacillus thuringiensis subsp. israelensis (Bti). The ppk gene of Bti was cloned into vector pHT315 and the effect of its overexpression on endotoxin production was determined. Endotoxin production by the recombinant strain was found to be consistently higher than that by the wild type strain and the strain that carried the empty plasmid. The toxicity of the recombinant mutant strain (LC50 5.8±0.6ngml(-1)) against late 2nd instar Culex quinquefasciatus was about 7.7 times higher than that of Bti (LC50 44.9±7ngml(-1)). To our knowledge this is the first reported study which relates polyP metabolism with bioinsecticide biosynthesis.