Possible effects of interleukin‐6 (IL‐6) on reproductive performance, embryonal development, parturition, and postnatal development have been suggested based on protein/mRNAexpression level of IL‐6 in related organs, but less is known about functions of IL‐6 signals in these areas. Following two different approaches have been employed to investigate the role of IL‐6 signals in fertility and pre‐/postnatal development: administration of a rat anti‐mouse IL‐6 receptor antibody, MR16‐1, to mice as a neutralizing antibody system, and B6.129S2‐Il6tm1Kopf/J (IL‐6 knockout [KO]) mice as a KO system. By intravenously dosing 50 mg/kg of MR16‐1 every 3 days, animals in male and female fertility studies and dams in a pre‐/postnatal development study exhibited plasma MR16‐1 concentrations much higher than the effective plasma concentration, indicating that MR16‐1 exposure was sufficient to completely block IL‐6 signals. The concentration of MR16‐1 in the plasma of fetuses exceeded that in the plasma of pregnant animals, and MR16‐1 concentration in milk was about one‐fourth of that in plasma. Both the transient IL‐6 signal blockade by MR16‐1, and the constitutive IL‐6 signal inhibition using IL‐6 KO mice in a combined fertility and pre‐/postnatal development study, revealed no biologically important effects on fertility, early embryonic development to implantation, or pre‐/postnatal development, including IgG/IgM production by keyhole limpet hemocyanin sensitization. These results indicate that IL‐6 signals have no unique, noncompensable roles in reproduction and development in the whole body system, although contributions of IL‐6 in the signaling network appear to exist, as suggested by previously published investigations.

INTRODUCTIONAs part of a collaborative study on toxicity related to female fertility, we performed experiments using di-(2-ethylhexyl) phthalate (DEHP), widely used as a plasticizer of polyvinyl chloride (PVC) and a known peroxisome proliferator (Lake et al., 1987), liver carcinogen (Reddy and Lalwai, 1983) and ovarian toxicant in rodents (Davis et al., 1994). Short-term exposure to DEHP resulted in hypoestrogenic anovulatory cycles and polycystic ovaries in adult rats (Davis et al., 1994). Longterm exposure to DEHP resulted in continuous diestrous stage with reduced serum estradiol, follicle stimulating hormone (FSH), pituitary FSH and luteinizing hormone (LH) (Hirosawa et al., 2006). The active metabolite of DEHP mono-(2-ethylhexyl) phthalate (MEHP) decreased aromatase in rat granulosa cells in vitro (Davis et al., 1994). Because the toxic effects of DEHP on ovaries are thought to be endocrine disorders and an aromatase inhibitor is expected to impact both ovarian morphology and female fertility, we selected DEHP as the test compound for the evaluation of ovarian toxicity.The present study was designed to determine whether 2-or 4-week administration periods for detecting histopathological changes in a repeated-dose toxicity study to compare the potential ovarian toxicity with the female reproductive function in a fertility study. MATERIALS AND METHODS Test articleDEHP was obtained from Kanto Chemical (Tokyo, Japan). Corn oil was purchased from Sigma-Aldrich Japan (Tokyo, Japan) and used as a vehicle. DEHP was ABSTRACT -The main purpose of this collaborative work is to determine the optimal administration period required to detect toxic effects in evaluation of ovarian morphological changes in repeated-dose toxicity studies. To assess the morphological and functional changes induced in the ovaries by di-(2-ethylhexyl) phthalate (DEHP), two repeated-dose toxicity studies (repeated dose for 2 or 4 weeks) of DEHP administrated to female rats at dose levels of 0, 300, 1,000 and 3,000 mg/kg were conducted in collaboration with a female fertility study at the same dosages from 2 weeks prior to mating to Day 7 of pregnancy. Histopathology of the ovaries in both repeated-dose toxicity studies showed vacuolation of stromal cells in the groups receiving 300 mg/kg or more and an increase of large atretic follicles in groups at 1,000 mg/ kg or more. In the 4-week study, a decrease in new corpora lutea was observed in the 3,000 mg/kg group. In the female fertility study, the 3,000 mg/kg group showed prolongation of the mean estrous cycle and irregular estrous cycles. Cesarean section revealed a decrease of pregnancy rate in the 3,000 mg/kg group. No effects on fertility or early embryonic development were found in groups at 1,000 mg/kg or less. -ation of drugs which induce ovarian damage. In conclusion, for a repeated-dose toxicity study, a 2-week

Background Although COVID-19 severity in cancer patients is high, the safety and immunogenicity of the BNT162b2 mRNA COVID-19 vaccine in patients undergoing chemotherapy for solid cancers in Japan have not been reported. Methods We investigated the safety and immunogenicity of BNT162b2 in 41 patients undergoing chemotherapy for solid cancers and in healthy volunteers who received 2 doses of BNT162b2. We evaluated serum IgG antibody titers for S1 protein by ELISA at pre-vaccination, prior to the second dose and 14 days after the second vaccination in 24 cancer patients undergoing cytotoxic chemotherapy (CC group), 17 cancer patients undergoing immune checkpoint inhibitor therapy (ICI group) and 12 age-matched healthy volunteers (HV group). Additionally, inflammatory cytokine levels were compared between the HV and ICI groups at pre and the next day of each vaccination. Results Anti-S1 antibody levels were significantly lower in the ICI and CC groups than in the HV group after the second dose (median optimal density: 0.241 [0.063–1.205] and 0.161 [0.07–0.857] vs 0.644 [0.259–1.498], p = 0.0024 and p < 0.0001, respectively). Adverse effect profile did not differ among the three groups, and no serious adverse event occurred. There were no differences in vaccine-induced inflammatory cytokines between the HV and ICI groups. Conclusion Although there were no significant differences in adverse events in three groups, antibody titers were significantly lower in the ICI and CC groups than in the HV group. Further protection strategies should be considered in cancer patients undergoing CC or ICI.

-Totally implantable catheter animal models are considered useful for pharmacological and toxicological studies. In this report, we assessed the feasibility of using an indwelling vascular access port (VAP) in rats for long-term evaluation of repeated and intermittent dose toxicity studies. In Experiment 1, the VAP devices were implanted in male and female rats and a saline solution administered intravenously via the posterior vena cava for 2 weeks (4 ml/kg, 2 ml/min, 5 times/week, 10 times total). General conditions, body weight and blood chemistry showed no toxicological changes compared with the rats in the non-implanted, non-treated group. Hematology changes such as transient increases in peripheral blood reticulocytes and eosinophils were noted post-implantation. In pathology, proliferation of the -nophils in lung were noted at the end of the treatment period. Moreover, we found that the lumbar area is more suitable for VAP implantation than the back of neck for young, still growing rats. Experiment 2 included a 1-month intravenous intermittent dose (4 ml/kg, 2 ml/min, 1 time/week, 5 times total) toxicity study in VAP-implanted rats followed by a 1-month recovery period conducted under Good Laboratory Practice (GLP) regulations. The results suggested that an animal model with implanted VAP is useful for intermittent intravenous dosing of drugs. Moreover, VAP implantation in animals is expected to be extrapolated to use VAP in humans in clinical studies.

This study was conducted to evaluate the feasibility of total parenteral nutrition (TPN) in rats using continuous intravenous infusion (tail cuff method) via the posterior vena cava. A catheter was inserted into the posterior vena cava from the femoral vein of 10 females (Experiment 1) and 16 females (Experiment 2). The depth of the inserted catheter from the femoral vein was set at 4.5 cm for Experiment 1 and was set at 6 cm for Experiment 2. The test animals were divided into two groups in each experiment: a 5% D-Mannitol (MAN) group and a TPN group. In Experiment 1, TPN rats showed macroscopic lesions (edema in peritoneum, increased collateral vasculature, induration in perivenous tissue, and thrombus) at the tip of the catheter. The diameter of the posterior vena cava (2.86 +/- 0.16 mm, mean +/- S.D.) was significantly greater than that of the anterior vena cava (2.45 +/- 0.22 mm) in 10 rats of Experiment 1. In Experiment 2, TPN rats showed no abnormalities at necropsy. Our findings suggest that TPN administered via the posterior vena cava in Sprague-Dawley rats requires the catheter to be inserted to a depth of 6 cm from the femoral vein. We hypothesize that this is because it is inserted to the level of the renal vein branch where the diameter of the posterior vena cava may be greatest.