The Effects of GSPT1 Degradation on Serum Calcium, Parathyroid Hormone, and Fibroblast Growth Factor 23 Concentrations in Human Cereblon Knock-in Mice

Test article CC-325 is a potent oral cereblon (CRBN) modulator that has shown potent G1 to S phase transition 1 (GSPT1) degradation and anti-tumor activity in pre-clinical models. One of the adverse effects associated with CC-325 was dose dependent hypocalcemia, which was determined to be an on-target toxicity. To investigate the mechanism of hypocalcemia, we conducted a toxicity study in human cereblon (huCRBN) knock-in (KI) mice with CC-325. The huCRBN KI mice are transgenic mice engineered to express human cereblon that is capable of binding to CRBN and degrading GSPT1. Four groups of mice were treated with vehicle (0 mg/kg), CC-325 (50 mg/kg BID), NPS 2143 (120 mg/kg), or CC-325 + NPS 2143. The NPS 2143 is an oral negative allosteric modulator of calcium sensing receptor (CaSR), which upon administration to mice significantly increased plasma ionized calcium (iCa2+) and parathyroid hormone (PTH). Mice treated with CC-325 alone had significant decreases in serum iCa2+ and PTH, while mice treated with NPS 2143 alone as expected had significant increases in serum iCa2+ and PTH. Treatment of mice with CC-325 + NPS 2143 did not reverse the decreases in serum iCa2+ and PTH caused by CC-325, indicating that CC-325 prevents the increase of PTH. To investigate the mechanism of hypocalcemia, we stained parathyroid gland for PTH by immunohistochemistry (IHC) and showed significantly lower PTH in parathyroid with CC-325 treated mice compared to vehicle or NPS 2143 treated mice. To further investigate the cause of low PTH in parathyroid gland in mice treated with CC-325, we stained parathyroid with in-situ hybridization (ISH) probes for PTH mRNA. Results from this analysis showed significantly lower PTH mRNA in parathyroid of CC-325 mice compare to vehicle or NPS 2143 mice, indicating that lower serum PTH in CC-325 treated mice were due to decreased PTH mRNA in Chief cells. These data collectively indicate that hypocalcemia caused by CC-325 is due to reduction in PTH, which leads to hypocalcemia. Additionally, mice treated with CC-325 are unable to restore normocalcemia because their parathyroid gland did not synthesize sufficient PTH for release into blood stream. Lack of PTH synthesis is caused by diminished level of PTH mRNA in parathyroid gland. We also measured the level of FGF23 in mice treated with CC-325. Our data indicated that decrease in PTH significantly decreased FGF23 levels even in presence of hyperphosphatemia, indicating that PTH plays a big role in controlling FGF23 during hypoparathyroidism. The cause of decrease in PTH mRNA in parathyroid, whether it is related to lower transcription of PTH mRNA or lack of stability of PTH mRNA, remains to be determined

The Combined Effect of Photobiomodulation and Curcumin on Acute Skin Wound Healing in Rats

Introduction: Abnormal wound repair is a cause for a considerable expense, as well as patient morbidity and mortality. Here, we investigated the combined impact of photobiomodulation (PBM) and curcumin on a rat experimental model of an acute skin wound.Methods: A round full-thickness wound was created on the back of each rat. We divided the rats into the following four groups. Group one was the control group. Group two received pulse wave (PW) PBM at a dose of 890 nm, 80 Hz, and 0.2 J/cm2. Group 3 received 40 mg/kg curcumin by gastric gavage and group 4 was treated with PWPBM + curcumin. We measured the wound area on days 4, 7, and 15, and performed microbiologically and tensiometry examinations.Results: There was markedly improved wound contraction in the curcumin (7.5 ± 0.57; P=0.000), PBM (8.5 ± 1.2; P=0.000), and PBM + curcumin (14.5 ± 4.3; P=0.002) groups relative to the control group (25 ± 6). PBM (100 ± 7.3; P=0.005), and PBM + curcumin (98 ± 6; P=0.005) groups meaningfully improved tensile strength relative to the control group (61 ± 8.2). On day 15, the PBM (10 ± 5; P=0.000), curcumin (14 ± 4.5, P=0.000), and PBM + curcumin (27.3 ± 8.3; P=0.000) groups meaningfully decreased microbial flora relative to the control group (95 ± 6).Conclusion: We concluded that the PBM and PBM + curcumin groups meaningfully accelerated wound healing of the acute skin wound in the rats. The results of the PBM group were statistically more effective than the curcumin alone and PBM + curcumin-treated groups. DOI:10.34172/jlms.2021.0

Evaluation of the Effects of Photobiomodulation on Partial Osteotomy in Streptozotocin-Induced Diabetes in Rats

Objective: We examined the effects of photobiomodulation (PBM) on stereological parameters, and gene expression of Runt-related transcription factor 2 (RUNX2), osteocalcin, and receptor activator of nuclear factor kappa-B ligand (RANKL) in repairing tissue of tibial bone defect in streptozotocin (STZ)-induced type 1 diabetes mellitus (TIDM) in rats during catabolic response of fracture healing. Background data: There were conflicting results regarding the efficacy of PBM on bone healing process in healthy and diabetic animals. Materials and methods: Forty-eight rats have been distributed into four groups: group 1 (healthy control, no TIDM and no PBM), group 2 (healthy test, no TIDM and PBM), group 3 (diabetic control, TIDM and no PBM), and group 4 (diabetic test, no TIDM and PBM). TIDM was induced in the groups 3 and 4. A partial bone defect in tibia was made in all groups. The bone defects of groups second and fourth were irradiated by a laser (890 nm, 80 Hz, 1.5 J/cm2 ). Thirty days after the surgery, all bone defects were extracted and were submitted to stereological examination and real-time polymerase chain reaction (RT-PCR). Results: PBM significantly increased volumes of total callus, total bone, bone marrow, trabecular bone, and cortical bone, and the numbers of osteocytes and osteoblasts of callus in TIDM rats compared to those of callus in diabetic control. In addition, TIDM increased RUNX2, and osteocalcin in callus of tibial bone defect compared to healthy group. PBM significantly decreased osteocalcin gene expression in TIDM rats. Conclusions: PBM significantly increased many stereological parameters of bone repair in an STZ-induced TIDM during catabolic response of fracture healing. Further RT-PCR test demonstrated that bone repair was modulated in diabetic rats during catabolic response of fracture healing by significant increase in mRNA expression of RUNX2, and osteocalcin compared to healthy control rats. PBM also decreased osteocalcin mRNA expression in TIDM rats

Effects of pentoxifylline and alendronate on fracture healing in ovariectomy-induced osteoporosis in rats

Osteoporosis is determined by decreased bone strength that increases the threat of fractures. The aim of this study was to evaluate the effects of pentoxifylline (PTX) and alendronate (ALN), on the stereological parameters, and gene expression in callus of fracture in an experimental rat model of ovariectomy-induced osteoporosis (OVX). The OVX was induced in 90 female rats. Fourteen weeks later, a complete fracture on the right femur was made. Rats were divided into five groups: 1) control: no treatment; 2) sham: received daily distilled water; 3) daily 3.00 mg kg(-1) ALN subcutaneously (SC); 4) daily 200 mg kg(-1) PTX (SC) and 5) daily PTX (SC) + ALN (same doses). The osteoclast count was significantly lower in all treatment groups, at 21 and 56 days post-surgery, compared to the control and sham groups. The PTX significantly increased total callus volume at 21 and 56 days post-surgery, compared to the other groups. The PTX+ALN treatment significantly increased both cortical bone volume on day 21, and osteocyte and osteoblast numbers on day 56, compared to the control and sham groups. It can be concluded that PTX and ALN have antiresorptive effects, in OVX rats. Also, PTX has increased the extracellular matrix on both 21 and 56 days after surgery, compared to the other groups. PTX+ALN elevated cortical bone volume on day 21, and osteocyte and osteoblast numbers compared to the control and sham groups on day 56. (C) 2019 Urmia University. All rights reserved

ハチスカケ カシンダン セイリツショ ノ メノト ロウジョ カンケイ シリョウ ニツイテ

Osteoporosis is determined by decreased bone strength that increases the threat of fractures. The aim of this study was to evaluate the effects of pentoxifylline (PTX) and alendronate (ALN), on the stereological parameters, and gene expression in callus of fracture in an experimental rat model of ovariectomy-induced osteoporosis (OVX). The OVX was induced in 90 female rats. Fourteen weeks later, a complete fracture on the right femur was made. Rats were divided into five groups: 1) control: no treatment; 2) sham: received daily distilled water; 3) daily 3.00 mg kg-1 ALN subcutaneously (SC); 4) daily 200 mg kg-1 PTX (SC) and 5) daily PTX (SC) + ALN (same doses). The osteoclast count was significantly lower in all treatment groups, at 21 and 56 days post-surgery, compared to the control and sham groups. The PTX significantly increased total callus volume at 21 and 56 days post-surgery, compared to the other groups. The PTX+ALN treatment significantly increased both cortical bone volume on day 21, and osteocyte and osteoblast numbers on day 56, compared to the control and sham groups. It can be concluded that PTX and ALN have antiresorptive effects, in OVX rats. Also, PTX has increased the extracellular matrix on both 21 and 56 days after surgery, compared to the other groups. PTX+ALN elevated cortical bone volume on day 21, and osteocyte and osteoblast numbers compared to the control and sham groups on day 56. Keywords Fracture healing Osteoporosis Ovariectomy Real time PCR Stereolog

Analysis of sexual dimorphism in the Persian long-tailed desert lizard, Mesalina watsonana (Stoliczka, 1872; Sauria: Lacertidae)

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Evaluation of the Effects of Photobiomodulation on Partial Osteotomy in Streptozotocin-Induced Diabetes in Rats

Objective: We examined the effects of photobiomodulation (PBM) on stereological parameters, and gene expression of Runt-related transcription factor 2 (RUNX2), osteocalcin, and receptor activator of nuclear factor kappa-B ligand (RANKL) in repairing tissue of tibial bone defect in streptozotocin (STZ)-induced type 1 diabetes mellitus (TIDM) in rats during catabolic response of fracture healing. Background data: There were conflicting results regarding the efficacy of PBM on bone healing process in healthy and diabetic animals. Materials and methods: Forty-eight rats have been distributed into four groups: group 1 (healthy control, no TIDM and no PBM), group 2 (healthy test, no TIDM and PBM), group 3 (diabetic control, TIDM and no PBM), and group 4 (diabetic test, no TIDM and PBM). TIDM was induced in the groups 3 and 4. A partial bone defect in tibia was made in all groups. The bone defects of groups second and fourth were irradiated by a laser (890nm, 80Hz, 1.5J/cm(2)). Thirty days after the surgery, all bone defects were extracted and were submitted to stereological examination and real-time polymerase chain reaction (RT-PCR). Results: PBM significantly increased volumes of total callus, total bone, bone marrow, trabecular bone, and cortical bone, and the numbers of osteocytes and osteoblasts of callus in TIDM rats compared to those of callus in diabetic control. In addition, TIDM increased RUNX2, and osteocalcin in callus of tibial bone defect compared to healthy group. PBM significantly decreased osteocalcin gene expression in TIDM rats. Conclusions: PBM significantly increased many stereological parameters of bone repair in an STZ-induced TIDM during catabolic response of fracture healing. Further RT-PCR test demonstrated that bone repair was modulated in diabetic rats during catabolic response of fracture healing by significant increase in mRNA expression of RUNX2, and osteocalcin compared to healthy control rats. PBM also decreased osteocalcin mRNA expression in TIDM rats

Combined Treatment of Photobiomodulation and Arginine on Wound Healing in an Animal Model of Delayed Healing Wound: Photobiomodulation and Arginine on Chronic Wound Healing

Introduction: Herein, the individual and combined effects of photobiomodulation (PBM) and arginine (ARG) on the wound healing course of an experimental model of a slow healing wound (ulcer) in rats were assessed. Methods: A total of 108 male rats were divided into 6 groups: control; lower energy density (low)-PBM; arginine ointment (ARG); low-PBM+ARG; high energy density (high)-PBM; and highPBM+ARG. In each rat, one ischemic wound in the center of a bipedicle flap and one non-ischemic wound out of the flap was created. Both wounds were treated in the experimental groups. Microbial growth wound area and wound strength were assessed on days 0, 5, 10, 15, and 20 after the wound.infliction Results: All non-ischemic wounds closed before day 15. High-PBM+ARG and ARG significantly increased wound closure rates compared to the control group (LSD test, P=0.000, and P=0.001, respectively) on day 10. All slow healing wounds were open on day 15 but closed completely before day 20. Low-PBM+ARG and high-PBM significantly increased wound strength (stress high load, SHL) on day 10 compared to the control group (LSD test, P=0.001, and P=0.000, respectively). ARG, high-PBM, and low-PBM+ARG significantly increased wound closure rates on day 15 relative to the control group (LSD test, P=0.000, P=0.000, and P=0.001, respectively). Conclusions: High-PBM and low-PBM+ARG have biostimulatory and antibacterial effects on slow-healing wounds, which were shown by significant increases in wound closure rates, wound strength, and inhibition of Staphylococcus aureus growth. Doi:10.34172/jlms.2021.40

Factors Affecting Oral Hygiene Behavior in 9-12-Year-Old Children Based on Data from Fars Birth Cohort Study

Background: Several factors may influence oral hygiene behavior, which is essential for oral health and well-being in children. This study aimed to investigate the factors that contribute to the oral hygiene behaviors in children, especially mothers’ oral hygiene behaviors.Methods: In this cross-sectional study, data from the fifth phase of Fars Birth Cohort Study were used. Our focus was on a group of 1143 children aged 9-12 years old in the year 2021. The demographic, socio-economic variables as well as oral hygiene behaviors of children and their mothers were assessed. The score for oral hygiene behavior was determined based on self-reported practices of brushing, flossing, and mouthwash, ranged between 0 and 3. The factors related to children’s oral hygiene behavior were examined by logistic regression analysis.Results: In this study, 345 (30.2%) of the children (50.3% females, average age 10.33±0.48 years) did not brush their teeth, 1042 (91.2%) did not use dental floss, and 1083 (94.8%) did not use mouthwash. The mean score for oral hygiene behavior in children was 0.83±0.67. Mothers had a higher mean score of 1.42±0.74 (P<0.001). Several factors were found to be significantly associated with oral hygiene behavior in children, including higher scores for mothers’ oral hygiene behavior (OR=2.18, P<0.001), living in the center of the province (OR=0.73, P=0.04), having insurance (OR=1.47, P=0.03), and having a government-employed mother (OR=0.52, P=0.025). Children with regular six-monthly or yearly visits had higher odds compared with those who visited the dentist only when a problem arose (OR=7.27, P=0.009, and OR=2.33, P=0.01, respectively).Conclusion: The study revealed unacceptable oral hygiene behaviors in 9-12-year-old children, and showed the gap between oral hygiene behavior in mothers and children. The mother’s role in promoting oral health and hygiene is crucial. Future research should consider mediating and confounding impacts of knowledge, attitudes, and practices related to oral hygiene

Improvement in infected wound healing in type 1 diabetic rat by the synergistic effect of photobiomodulation therapy and conditioned medium

We investigated the effects of photobiomodulation therapy (PBMT) and conditioned medium (CM) of human bone marrow mesenchymal stem cells (hBM-MSC) individually and/or in combination on the stereological parameters and the expression of basic fibroblast growth factor (bFGF), hypoxia-inducible factor (HIF-1α), and stromal cell–derived factor-1α (SDF-1α) in a wound model infected with methicillin-resistant Staphylococcus aureus (MRSA) in diabetic rats. CM was provided by culturing hBM-MSCs. Type 1 diabetes mellitus (T1DM) was induced in 72 rats, divided into four groups, harboring 18 rats each: group 1 served as a control group, group 2 received PBMT, group 3 received CM, and group 4 received CM + PBMT. On days 4, 7, and 15, six animals from each group were euthanized and the skin samples were separated for stereology examination and gene expression analysis by real-time polymerase chain reaction. In the CM + PBMT, CM, and PBMT groups, significant decreases were induced in the number of neutrophils (1460 ± 93, 1854 ± 138, 1719 ± 248) and macrophages (539 ± 69, 804 ± 63, 912 ± 41), and significant increases in the number of fibroblasts (1073 ± 116, 836 ± 75, 912 ± 41) and angiogenesis (15 230 ± 516, 13 318 ± 1116, 14 041 ± 867), compared with those of the control group (2690 ± 371, 1139 ± 145, 566 ± 90, 12 585 ± 1219). Interestingly, the findings of the stereological examination in the CM + PBMT group were statistically more significant than those in the other groups. In the PBMT group, in most cases, the expression of bFGF, HIF-1α, and SDF-1α, on day 4 (27.7 ± 0.14, 28.8 ± 0.52, 27.5 ± 0.54) and day 7 (26.8 ± 1.4, 29.6 ± 1.4, 28.3 ± 1.2) were more significant than those in the control (day 4, 19.3 ± 0.42, 25.5 ± 0.08, 22.6 ± 0.04; day 7, 22.3 ± 0.22, 28.3 ± 0.59, 24.3 ± 0.19) and other treatment groups. The application of PBMT + CM induced anti-inflammatory and angiogenic activities, and hastened wound healing process in a T1 DM model of MRSA infected wound