Effect of Epididymal Spermatozoa of Local Iraqi Goat on in vitro Fertilization and Evolution of Embryos.

In vitro fertilization (IVF) has been considered one of the greatest improvements in livestock science and production. Epididymal spermatozoon is a valuable gamete source obtained from slaughtered animals, and it has great importance in livestock IVF. This study aimed to create caprine embryos in Baghdad using local Iraqi goats. Ovaries were obtained from 50 female local goats at the Al-shu'alah abattoir. The ovaries were transported to the laboratory in physiological saline solution at 38°C. After oocyte recovery from the mature antral follicle, the oocytes with grades A or B were used for in vitro maturation (IVM). Moreover, 10 testicles were obtained from slaughtered adult goats, and the sperm was recovered from the caudal epididymis. All the stage-specific culture mediums were incubated at 39°C with 5% CO2 and 90% relative humidity. In this study, embryonic development on a 24-h basis was evaluated and in vitro culture media was replaced 50% of the volume every day for nine days with fresh media until an enlarged or hatched blastocyst appeared. The recorded data showed that the mean size of the large follicles was 6.23±1.34 mm, while the small follicles were in the mean size of 3.10±0.62. On the other hand, the results showed a significant increase in the oocyte recovery rate from the large size follicle (81.22%), compared to the small size follicle (62.67%). The recorded data showed that the total number of retrieved oocytes with grades A and B was 290 (79.5%) . The recorded data showed that 50.1% of these oocyte cohorts had reached metaphase II (145/290). Furthermore, 45.45% of the recovered oocytes by aspiration from large follicles were developed to the blastocyst stage, compared to 25% of recovered oocytes from the small follicles reaching the same developmental stage (blastocyst). On the other hand, the recorded data showed that the oocyte recovery method had a great influence on the oocyte competence regardless of the follicle size. The results revealed that the number of recovered oocytes by aspiration was greater than slicing. In conclusion, the results approved that the slicing method could be considered a safe and more efficient method in oocyte recovery from the slaughtered animals, compared to the aspiration method. On the other hand, according to the findings, the epididymal spermatozoa from local Iraqi goats are a reliable source of oocytes for IVF and in vitro conversion.

The solute carrier family 7 member 11 (SLC7A11) is regulated by LH/androgen and required for cystine/glutathione homeostasis in mouse Sertoli cells.

Luteinizing hormone (LH) stimulates testosterone production from Leydig cells. Both LH and testosterone play important roles in spermatogenesis and male fertility. To identify LH - and testosterone - responsive transporter genes that play key roles in spermatogenesis, we performed large-scale gene expression analyses on testes obtained from adult control and Lhb knockout mice. We found a significant reduction in cystine/glutamate transporter encoding Slc7a11 mRNA in testes of Lhb null mice. We observed that Slc7a11/SLC7A11 expression was initiated pre-pubertally and developmentally regulated in mouse testis. Immunolocalization studies confirmed that SLC7A11 was mostly expressed in Sertoli cells in testes of control and germ cell-deficient mice. Western blot analyses indicated that SLC7A11 was significantly reduced in testes of mutant mice lacking either LH or androgen receptor selectively in Sertoli cells. Genetic and pharmacological rescue of Lhb knockout mice restored the testicular expression of Slc7a11 comparable to that observed in controls. Additionally, Slc7a11 mRNA was significantly suppressed upon Sertoli cell/testicular damage induced in mice by cadmium treatment. Knockdown of Slc7a11 in vitro in TM4 Sertoli cells or treatment of mice with sulfasalazine, a SLC7A11 inhibitor caused a significant reduction in intracellular cysteine and glutathione levels but glutamate content remained unchanged as determined by metabolomic analysis. Knockdown of Slc7a11 resulted in compensatory upregulation of other glutamate transporters belonging to the Slc1a family presumably to maintain intracellular glutamate levels. Collectively, our studies identified that SLC7A11 is an LH/testosterone-regulated transporter that is required for cysteine/glutathione but not glutamate homeostasis in mouse Sertoli cells.

High intensity focused ultrasound as first line treatment in patients with chronic angle closure glaucoma at risk for malignant glaucoma.

PURPOSE: Evaluation of high intensity focused ultrasound (HIFU) transscleral cyclocoagulation as an alternative to trabeculectomy in the treatment of refractory chronic angle closure glaucoma (CACG). METHODS: This prospective one armed single center pilot study was conducted on patients with medically uncontrolled CACG who underwent high intensity focused ultrasound (HIFU) cyclocoagulation as first line surgical treatment, using the Eye-OP1 HIFU device (Eyetechcare-Rillieux-la-Pape, France) driven by ultrasound biomicroscopic (UBM) ciliary body localization. All patients had documented progression of their glaucoma despite conventional medical and laser therapy. The primary efficacy outcome was intraocular pressure (IOP) reduction; secondary outcomes included changes in number of glaucoma medications and complications after cyclocoagulation. RESULTS: The study involved 8 eyes of 7 patients between May 2013 and September 2014. The average follow-up was 5.6±2.1 months. The mean IOP was reduced from 18.4±3.5mmHg preoperatively to 14.8±4.1mmHg 6 months postoperatively. The average number of glaucoma medications decreased from 3.4±1.1 at baseline to 3.3±0.7 after 6 months. Visual acuity remained stable (median 0.17 log MAR preoperatively and 0.19 log MAR at last follow-up visit). No significant side effect occurred during the 6 months follow-up period. CONCLUSIONS: HIFU cyclocoagulation appears to be a safe and reliable alternative to filtering surgery in the management of chronic angle closure glaucoma among patients with a high risk of malignant glaucoma.

Ultrasonographic renal volume measurements in early autosomal dominant polycystic disease: comparison with CT-scan renal volume calculations.

PURPOSE: To investigate the correlation and concordance between the ellipsoid volume calculated by ultrasonography measurements (Vol3DUS) and the reference kidney volume measured by CT (VolTDM) in early autosomal dominant polycystic kidney disease (ADPKD). MATERIALS AND METHODS: Prospective study of the correlation and concordance of renal volumes in 24 patients with early ADPKD (48 kidneys analysed separately), with calculation of Vol3DUS using the formula for an ellipsoid in three different manners and VolTDM measurement by manual contouring. Calculations of correlation coefficients (r) and coefficients of intra-class correlation (ICC) with confidence intervals at 95%. RESULTS: The US volume was strongly correlated with the CT volume by using the maximum width in a transverse section (r=0.83) with a mean Vol3DUS=692±348ml [180; 2069]. The most reproducible ultrasonography measurement was the height. When the kidney volume exceeded 800ml, US underestimated the volume. However, the median error was -57.5ml [-1090; 183] and 85% of the Vol3DUS calculated differed by more than 5% from the reference measurement. CONCLUSION: The correlation between the US calculated volumes and the CT volumes was strong. However, the median error with ellipsoid US volume was too high to detect a small renal variation in early ADPKD.

Protamine ratio and the level of histone retention in sperm selected from a density gradient preparation.

Fertile males express two forms of sperm nuclear proteins, protamine 1 (P1) and protamine 2 (P2), in roughly equal quantities, whereas some infertile men have been shown to have a reduction in protamine content and an increase in the level of histones retained in mature sperm. In this study, we assessed histone and protamine levels in spermatozoa isolated from different layers of a density gradient centrifugation column to evaluate the nuclear protein content of the sperm population selected. Protamine levels were measured using acid gel electrophoresis and immunofluorescence, and the percentage of cells retaining histones was evaluated using aniline staining and immunofluorescence. Our data suggests that there is an inverse correlation between P1/P2 ratio and the level of histone expression in the different layers of the density gradient. Paradoxically, the 90% layer had a lower P1/P2 ratio, which corresponded with an increase in histone expression. It is concluded that although the sperm population selected in the 90% layer of the density gradient columns had a lower P1/P2 ratio, it was yet similar to the P1/P2 ratio observed in previously screened fertile donors.

Theoretical calculation of the low lying electronic states of the molecular ion RbH(+) with spin-orbit effects.

The potential energy has been calculated for the 42 lowest electronic states of symmetries Omega=12,32,52, for the molecular ion RbH(+). Using an ab initio method, the calculation is based on nonempirical pseudopotentials and parametrized [script-l]-dependent polarization potentials. Gaussian basis sets have been used for both atoms, and spin-orbit effects have been taken into account. The spectroscopic constants for 19 electronic states have been calculated by fitting the calculated energy values to a polynomial in terms of the internuclear distance r. The permanent dipole moment and the transition dipole moments have been calculated for the considered Omega states. Through the canonical functions approach the eigenvalue E(v), the abscissas of the corresponding turning points (r(min) and r(max)) and the rotational constants B(v) have been calculated. The comparison of the present results with those available in literature shows a very good agreement.

Ewing's sarcoma presenting as a solitary cyst.

This case describes a 10-year-old girl who developed a Ewing's sarcoma in her proximal fibula. The radiologic features mimicked those of a unicameral bone cyst. The presence of pain and the atypical location led to a prompt biopsy and the correct diagnosis. The mechanism of this unusual radiographic presentation is discussed.