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This study aimed to evaluate the effect of adding insulin hormone at different levels (0, 5, 10 and 15 µg/ml) to maturation medium with or without exogenous hormones (FSH, LH and E17β) on the in vitro maturation (IVM) of rabbit oocytes. Total of 20 New Zealand White (NZW) rabbit does (5.5-6 mo of age and 2.5-3 kg LBW) were used as oocyte donors. Oocytes were recovered from ovaries of slaughtered does using slicing technique. All oocytes with evenly granulated dark ooplasm were matured in TCM-199 supplemented with 6% bovine serum albumin. Eight types of TCM-199, four types without and other four with exogenous hormones supplemented with insulin (0, 5, 10 and 15 µg /ml) were used. Oocytes were fixed and stained for examination after 18 h at 38.5°C, 5% CO2 and high humidity as a maturation period. Results showed significant (P<0.05) effect of insulin supplementation on IVM of rabbit oocyte only in terms of oocytes reaching MI, TI+MII and degenerated ones. Percentage of mature oocytes reaching MII was improved by all insulin levels as compared to un-supplemented media (42.1-44.6 vs. 34.1%), but the differences were not significant. Percentage of oocytes reaching both TI+MII increased (P<0.05) with insulin supplementation at a level of 5 µg/ml showing the best (P<0.05) improvement on oocyte maturation (51.0%) as compared to other insulin levels (45.5-47.6%) or the control medium (39.5%). Nuclear maturation of rabbit oocytes was not affected significantly when FSH, LH and E2 were deleted from the maturation medium, but Percentage of oocytes reaching both T1+MII stages was enhanced in the presence of hormones during the maturation period regardless of whether oocytes were treated with insulin or not. Maturation rate in term of oocytes reaching T1+MII was affected significantly (P<0.05) by the interaction between insulin and hormonal addition, reflecting improved percentage of oocytes reaching both T1+MII stages by addition of all insulin levels to hormone-TCM-199 medium. In this respect, insulin addition at a level of 5µg/ml showed the best result (58.7%). In conclusion, the present study demonstrated that the supplementation of the maturation medium with insulin improves the in vitro maturation rate of rabbit oocytes when oocytes are maturated in a defined maturation medium with or without hormones (FSH, LH and E2).

Background The severity of respiratory failure in SARS-CoV-2 infection appears to be related to cytokine release syndrome (CRS), resulting in mechanical ventilation (MV). In this observational study, we investigated tocilizumab’s role in the treatment of SARS-CoV-2 and the use of interleukin-6 (IL-6) levels in the management of these patients. Methods Patients with positive SARS-CoV-2 PCR were prospectively observed from February 1, 2020 to May 31, 2020. Data on demographics, medical history, and clinical outcomes were collected. Tocilizumab (TCZ) 4 mg/kg/day q12h was given for 24 hours, followed by methylprednisolone 60 mg q8h for 72 hours to patients with oxygen requirement of 3 L and above. IL-6, C-reactive protein (CRP), ferritin, lactate dehydrogenase (LDH), and D-dimers were monitored on days: 0, 3, and 6 following the initiation of therapy. Statistical analyses were performed using a Wilcoxon signed-rank test with significance (α) less than or equal to 0.05 (P ≤ 0.05). Results A total of eighty patients (45 males, 56.96%) and (34 females, 43.04%) with positive SARS-CoV-2 PCR were included in this study. The median age was 63 (51 - 72) years. Seven patients expired (8.75%), and nine patients required mechanical ventilation (11.25%). The median of IL-6 levels before administration of TCZ was 342.50 (78.25 – 666.25) pg/mL compared to after administration of TCZ on day 3, 563 (162-783) pg/mL (P &lt; 0.00001). On day 6, the median dropped to 545 (333.50 - 678.50) pg/mL as compared to day 3 (P = 0.709). Moreover, CRP, ferritin, LDH, and D-dimers levels were reduced following the administration of TCZ. TABLE 1: IL-6 of SARS-CoV-2 patients at before and after Tocilizumab treatment TABLE 2: Laboratory findings of SARS-CoV-2 patients at before and after Tocilizumab treatment Conclusion Early use of TCZ may reduce the need for MV and decrease CRP, ferritin, LDH, and D-dimer levels, which may be useful inflammatory indices in the management of SARS-CoV-2 patients. Furthermore, the sequential use of methylprednisolone for 72 hours seems to potentiate the effect and prolong the suppression of the cytokine storm. The use of IL-6 levels may be helpful as a prognostic tool and in the management of acutely ill SARS-CoV-2 patients. Disclosures All Authors: No reported disclosures