The impact of Apis dorsata forest honey administration on follicle-stimulating hormone and luteinizing hormone levels in rats (Rattus norvegicus) forced swim test as a chronic physical animal model.

Background: Chronic physical stress has many effects on the nervous system and can cause structural changes in different parts of the brain and hemomodulatory, including hormonal. Current pharmacotherapeutic treatments have limited efficacy and are associated with many deleterious side effects. Aim: The aim of this research is to determine how Apis dorsata forest honey administration affects follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels in rats who are subjected to forced swim tests as a model of chronic physical stress placed in a container filled with water from which it cannot escape. Methods: This was an experimental laboratory study with 32 rats divided into four treatment groups: control (C), Treatment 1 (T1) with a forced swim test + honey (2 g/rat/day), Treatment 2 (T2) with a forced swim test + honey (4 g/rat/day), and Treatment 3 (T3) with a forced swim test + honey (6 g/rat/day). All treatments were administered for 14 days. Then, blood was taken for FSH and LH serum tests, and a one-way ANOVA and Duncan test were used to statistically test the data analysis. Results: The results of this study indicate that the administration of forest honey had no significant effect (p > 0.05) on the FSH parameter, but there was a significant decrease in LH levels in the T2 and T3 groups (p < 0.05). Conclusion: It can be concluded that giving forest honey to rats who were subjected to a 14-day forced swim test had no effect on FSH and LH levels. In rats given a forced swim test as a model of chronic stress, administration at doses of 4 and 6 g/rat/day reduced LH serum levels. Thus, giving forest honey could maintain reproductive health in rat that experience chronic stress.

Mesenchymal stem cell secretome ameliorates over-expression of soluble fms-like tyrosine kinase-1 (sFlt-1) and fetal growth restriction (FGR) in animal SLE model.

INTRODUCTION: In the near future, stem cell research may lead to several major therapeutic innovations in medical practice. Secretome, a "by-product" of stem cell line cultures, has many advantages. Its easiness of storage, usage, and fast direct effect are some of those to consider. Fetal growth restriction (FGR) remains one of the significant challenges in maternal-fetal and neonatal medicine. Placentation failure is one of the most profound causal and is often related to increasing sFlt-1 in early pregnancy. This study aimed to investigate hUC-MSC secretome in ameliorating sFlt-1 and how to improve outcomes in preventing FGR in an animal model. MATERIALS AND METHODS: Pristane-induced systemic lupus erythematosus (SLE) in a mouse model was used to represent placentation failure and its consequences. Twenty-one mice were randomized into three groups: (I) normal pregnancy, (II) SLE, and (III) SLE with secretome treatment. Pristane was administered in all Groups four weeks prior mating period. Secretome was derived from human umbilical cord mesenchymal stem cells (hUC-MSC) conditioned medium on the 3rd and 4th passage, around day-21 until day-28 from the start of culturing process. Mesenchymal stem cell was characterized using flow cytometry for CD105+, CD90+, and CD73+ surface antigen markers. Immunohistochemistry anlysis by using Remmele's Immunoreactive Score (IRS) was used to quantify the placental sFlt-1 expression in each group. Birth weight and length were analyzed as the secondary outcome. The number of fetuses obtained was also calculated for pregnancy loss comparison between Groups. RESULTS: The administration of secretome of hUC-MSC was found to lower the expression of the placental sFlt-1 significantly in the pristane SLE animal model (10.30 ± 1.40 vs. 4.98 ± 2.57; p < 0.001) to a level seen in normal mouse pregnancies in Group I (3.88 ± 0.49; p = 0.159). Secretome also had a significant effect on preventing fetal growth restriction in the pristane SLE mouse model (birth weight: 354.29 ± 80.76 mg vs. 550 ± 64.03 mg; p < 0.001 and birth length: 14.43 ± 1.27 mm vs. 19.00 ± 1.41 mm), comparable to the birth weight and length of the normal pregnancy in Group I (540.29 ± 75.47 mg and 18.14 ± 1.34 mm, p = 0.808 and = 0.719). Secretome administration also showed a potential action to prevent high number of pregnancy loss as the number of fetuses obtained could be similar to those of mice in the normal pregnant Group (7.71 ± 1.11 vs. 7.86 ± 1.06; p = 0.794). CONCLUSIONS: Administration of secretome lowers sFlt-1 expression in placenta, improves fetal growth, and prevents pregnancy loss in a mouse SLE model.

Ethanol extract of basil (Ocimum Basilicum L.) leaves inhibits endometriosis growth in a mouse model by modulating vascular endothelial growth factor (VEGF) expression.

The objective of this study was to examine the effect of administering an ethanol extract obtained from basil leaves on the expression of vascular endothelial growth factor (VEGF) and the severity of endometriosis lesions in a mouse model. A total of 28 female mice, aged 2-3 months and weighing 20-30 grams, were randomly divided into four groups: the control group (C), treatment group 1 (T1) receiving a dose of basil leaf ethanol extract (0.21 mg/g-BW), treatment group 2 (T2) receiving a higher dose (0.42 mg/g BW), and treatment group 3 (T3) receiving the highest dose (0.84 mg/g-BW). Each group underwent a 14-day treatment period, and tissue samples were collected on the 29th day. An immunohistochemical examination was conducted to assess the expression of VEGF and evaluate the severity of endometriosis lesions. The statistical analysis of VEGF expression revealed a significant difference (p=0.026; p<0.05), with the most pronounced effects observed when administering basil leaf ethanol extract at doses of 0.21 mg/g-BW and 0.42 mg/g-BW. Although not statistically significant (p=0.271; p<0.05), a reduction in the severity of endometriosis lesions was observed following the administration of basil leaf ethanol extract at doses of 0.21 mg/g-BW and 0.42 mg/g-BW. Administering basil leaf ethanol extract at doses of 0.21 mg/g-BW and 0.42 mg/g-BW effectively decreased VEGF expression and limited the severity of endometriosis lesions.

Designing one-step reverse transcriptase loop-mediated isothermal amplification for serotype O foot-and-mouth disease virus detection during the 2022 outbreak in East Java, Indonesia.

Background and Aim: Various methods can detect foot-and-mouth disease (FMD) in cows, but they necessitate resources, time, costs, laboratory facilities, and specific clinical specimen submission, often leading to FMD virus (FMDV) diagnosis delays. The 2022 FMD outbreak in East Java, Indonesia, highlighted the need for an easy, inexpensive, rapid, and accurate detection approach. This study aims to devise a one-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) technique and phylogenetic analysis to detect the serotype O FMDV outbreak in East Java. Materials and Methods: Swab samples were collected from the foot vesicles, nasal secretions, and saliva of five suspected FMDV-infected cows in East Java between June and July 2022. The RT-LAMP design used hydroxy naphthol blue dye or SYBR Green I dye, with confirmatory analysis through reverse transcriptase polymerase chain reaction (RT-PCR) targeting 249 base pairs. PCR products underwent purification, sequencing, and nucleotide alignment, followed by phylogenetic analysis. Results: The RT-LAMP method using hydroxy naphthol blue dye displayed a positive reaction through a color shift from purple to blue in the tube. Naked-eye observation in standard light or ultraviolet (UV) light at 365 nm, with SYBR Green I stain, also revealed color change. Specifically, using SYBR Green I dye, UV light at 365 nm revealed a color shift from yellow to green, signifying a positive reaction. Nucleotide alignment revealed mutations and deletion at the 15th sequence in the JT-INDO-K3 isolate from the East Java FMDV outbreak. Despite differing branches, the phylogenetic tree placed it in the same cluster as serotype O FMDV from Malaysia and Mongolia. Conclusion: JT-INDO-K3 exhibited distinctions from Indonesian serotype O FMDV isolates and those documented in GenBank. Then, the RT-LAMP method used in this study has a detection limit 10 times higher latter than the conventional RT-PCR limit, without any cross-reactivity among strains.

Effect of Endometriosis on Cumulus ATP, Number of Mitochondria and Oocyte Maturity in Cumulus Oocyte Complex in Mice.

OBJECTIVE: Endometriosis causes a decrease in oocyte quality. However, this mechanism is not fully understood. The present study aimed to analyze the effect of endometriosis on cumulus cell adenosine triphosphate ATP level, the number of mitochondria, and the oocyte maturity level. METHODS: A true experimental study with a post-test only control group design on experimental animals. Thirty-two mice were divided into control and endometriosis groups. Cumulus oocyte complex (COC) was obtained from all groups. Adenosine triphosphate level on cumulus cells was examined using the Elisa technique, the number of mitochondria was evaluated with a confocal laser scanning microscope and the oocyte maturity level was evaluated with an inverted microscope. RESULTS: The ATP level of cumulus cells and the number of mitochondria in the endometriosis group increased significantly (p < 0.05; p < 0.05) while the oocyte maturity level was significantly lower (p < 0.05). There was a significant relationship between ATP level of cumulus cells and the number of mitochondrial oocyte (p < 0.01). There was no significant relationship between cumulus cell ATP level and the number of mitochondrial oocytes with oocyte maturity level (p > 0.01; p > 0.01). The ROC curve showed that the number of mitochondrial oocytes (AUC = 0.672) tended to be more accurate than cumulus cell ATP level (AUC = 0.656) in determining the oocyte maturity level. CONCLUSION: In endometriosis model mice, the ATP level of cumulus cells and the number of mitochondrial oocytes increased while the oocyte maturity level decreased. There was a correlation between the increase in ATP level of cumulus cells and an increase in the number of mitochondrial oocytes.

Effect of Endometriosis on Cumulus ATP, Number of Mitochondria and Oocyte Maturity in Cumulus Oocyte Complex in Mice / Efeito da endometriose no cumulus ATP, número de mitocôndrias e maturidade oocitária no complexo oocitário cumulus em camundongos

Abstract Objective Endometriosis causes a decrease in oocyte quality. However, this mechanism is not fully understood. The present study aimed to analyze the effect of endometriosis on cumulus cell adenosine triphosphate ATP level, the number of mitochondria, and the oocyte maturity level. Methods A true experimental study with a post-test only control group design on experimental animals. Thirty-two mice were divided into control and endometriosis groups. Cumulus oocyte complex (COC) was obtained from all groups. Adenosine triphosphate level on cumulus cells was examined using the Elisa technique, the number of mitochondria was evaluated with a confocal laser scanning microscope and the oocyte maturity level was evaluated with an inverted microscope. Results The ATP level of cumulus cells and the number of mitochondria in the endometriosis group increased significantly (p < 0.05; p < 0.05) while the oocyte maturity level was significantly lower (p < 0.05). There was a significant relationship between ATP level of cumulus cells and the number of mitochondrial oocyte (p < 0.01). There was no significant relationship between cumulus cell ATP level and the number of mitochondrial oocytes with oocyte maturity level (p > 0.01; p > 0.01). The ROC curve showed that the number of mitochondrial oocytes (AUC = 0.672) tended to be more accurate than cumulus cell ATP level (AUC = 0.656) in determining the oocyte maturity level. Conclusion In endometriosis model mice, the ATP level of cumulus cells and the number of mitochondrial oocytes increased while the oocyte maturity level decreased. There was a correlation between the increase in ATP level of cumulus cells and an increase in the number of mitochondrial oocytes.

Experimental Analysis of Vaginal Laxity in Rats Treated With a Combination of Er:YAG Fractional Lasers and AMSC-MP.

Introduction: Vaginal laxity, a symptom of pelvic floor dysfunction observed in women, has many negative biological and psychological impacts. Laser treatments and stem cell-based therapies are emerging therapeutic methods for treating this condition. This study aimed to determine changes in vaginal laxity in model rats using a combination therapy of erbium-doped yttrium aluminium garnet (Er:YAG) fractional lasers and topical treatment with amniotic membrane stem cell metabolite products (AMSC-MP). Methods: The experimental animal population comprised 36 female white rats (Rattus norvegicus; 2-day-post-vaginal-delivery rats) allocated into the following four groups (n=9): K1, untreated two-day-post-vaginal-delivery rats; K2, two-day-post-vaginal-delivery rats treated with topical gel without AMSC-MP; P1, two-day-post-vaginal-delivery rats treated with Er:YAG fractional lasers and topical gel without AMSC-MP; P2, two-day-post-vaginal-delivery rats treated with Er: YAG fractional lasers and topical gel containing AMSC-MP. Immunohistochemical (IHC) examination was carried out for the expression and activity of heat shock protein 70 (HSP-70), collagen-1, tissue inhibitors of metalloproteinase 1 (TIMP-1) and matrix metalloproteinase 1 (MMP-1), as well as vaginal mucosal thickness. Results: There was a significant difference (P<0.05) in the expression of HSP-70 among all groups except K2 and P1 (P>0.05); there was no significant difference in type I collagen and TIMP-1 expression between the groups (P>0.05); there was a significant difference (P<0.05) in MMP-1 activity, with the activity in the K2 group (5.79±0.83) being higher than that in the P1 group (4.44±1.82) and that in the K1 group (5.74±1.03) being higher than that in the P2 group (4.24±1.55). Also, there was a significant difference in the thickness of the vaginal mucosa in all groups except K2 and P1 (P>0.05). Conclusion: Er:YAG fractional laser and AMSC-MP combination therapy improved vaginal laxity in model rats by increasing Hsp70 expression and vaginal mucosal thickness and decreasing MMP-1 activity.

Correlation of serum kisspeptin levels, ovarian kisspeptin expression, and ovarian BMP15 expression in rat model of polycystic ovary syndrome.

Background: Kisspeptin is a neuropeptide that has an important role in the female reproductive cycle which is indicated by its role in regulating the hypothalamic-pituitary-gonadal axis. Aims: To analyze the correlation between serum kisspeptin levels, ovarian kisspeptin expression, and ovarian Bone Morphogenic Protein-15 (BMP15) expression in polycystic ovary syndrome (PCOS) model rats. Methods: The research was accurate experimental research with a post-test design-only control group and was carried out from August to October 2022 at the Faculty of Veterinary Medicine Universitas Airlangga. 32 Rattus novergicus rats were divided into a control group and a PCOS model group. Blood serum and ovaries were obtained from all groups. In addition, blood serum was examined for kisspeptin levels by ELISA technique, and kisspeptin expression and BMP15 Ovaries were examined immunohistochemically. Results: Serum kisspeptin levels and ovarian kisspeptin expression of the PCOS model group were not significantly higher than those of the control group (p > 0.05, p > 0.05). The ovarian BMP15 expression of the PCOS model group was not significantly lower (p > 0.05) than that of the control group. Ovarian kisspeptin expression and ovarian BMP15 expression did not significantly correlate with serum kisspeptin levels (p > 0.05). In contrast, there was a significant correlation (p < 0.05) between ovarian kisspeptin expression and ovarian BMP15 expression. Conclusion: Serum kisspeptin levels and ovarian kisspeptin expression of the PCOS model group were not higher than those of the control group, and the ovarian BMP15 expression of the PCOS model group was not lower than that of the control group. There was no correlation between serum kisspeptin levels with ovarian kisspeptin expression and ovarian BMP15 expression. However, a significant correlation was found between ovarian kisspeptin expression and ovarian BMP15 expression.

Curcuma longa supplementation reduces MDA, TNF-α, and IL-6 levels in a rat model exposed to soot particulates.

Background: Particulate matter (PM) is one of the important components in air pollution that can cause endothelial vascular dysfunction through exacerbation of atherosclerosis and inflammation of the respiratory system. Increased levels of malondialdehyde (MDA) in blood plasma can be an indicator of oxidative stress. Then, macrophages can secrete proinflammatory cytokines that will stimulate immune cells and vascular endothelial cells to release inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α). Curcuma longa works by scavenging the active free radicals involved in the peroxidation process. Aims: This study aims to prove that the administration of C. longa can reduce MDA, TNF-α, and IL-6 levels in Rattus norvegicus exposed to soot particulates. Methods: The subjects of this study were 30 male rats which were divided into 5 treatment groups with the following: (C0): negative control; (C+): positive control; (T1): Treatment group 2, rats exposed to particulate soot at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 1 mg/kg bw; (T2): Treatment group 3 was rats exposed to soot particulates at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 2 mg/kg bw; (T3): Treatment group 4 was rats exposed to soot particulates at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 3 mg/kg bw.Giving the C. longa extract orally with a probe every day for 30 days after treatment of exposure to soot. Examination of MDA, TNF-, and IL-6 levels with the ELISA method. Results: The administration of C. longa can reduce MDA while the lowest MDA levels were obtained in the T3 treatment with an average of 1.542 ± 0.231. The results of the description of the lowest levels of TNF-α were obtained in the C-treatment with an average of 55.981 ± 4.689. Then, the lowest levels of IL-6 were obtained in the C-treatment with an average of 2.292 ± 0.461. Conclusion: The results stated that the administration of C. longa could reduce MDA levels, TNF-α, and IL-6 levels. Curcuma longa as an anti-inflammatory and anti-oxidant play an effective role in inhibiting inflammation by decreasing IL-6 cytokine and TNF-α. Curcuma longa can inhibit lipid peroxidation initiated by free radicals and then reduce MDA levels.

The Impact of Endometriosis on Intracellular Calcium Levels, Cyclic Dependent Kinase 1 (Cdk1) Expression, and Cyclin B Expression in Post-Ovulation Oocytes of Mice Model.

Background: Since endometriosis causes a decrease in oocyte quality, the success rate of in vitro fertilization cycles decreases. The purpose of the current study was to analyze the effect of endometriosis on intracellular calcium levels, Cdk1 expression, and cyclin B expression in oocytes. Methods: Thirty-two mice (Mus musculus) were divided into control and endometriosis groups. The cumulus oocyte complex (COC) were obtained in all groups. Denudated cells were assessed for calcium levels by calorimetric examinations. Complex oocytes were examined for Cdk1 and cyclin B expression by immune-cytochemistry and were read under a microscope. Results: Intercellular calcium levels, Cdk1, and cyclin B expression were significantly lower in the endometriosis group than in the control group. There was a significant relationship between calcium levels and Cdk1 expression (p<0.05, r=0.659), a significant relationship between calcium levels and cyclin B expression (p<0.05, r=0.885), and also a significant correlation between Cdk1 and cyclin B expression (p<0.05, r=0.537). Conclusion: The data presented in this study suggested that the intracellular oocyte calcium level, Cdk1 expression, and cyclin B expression were lower in mice with endometriosis. A positive correlation was observed between calcium levels and the expression of Cdk1 and cyclin B. Furthermore, a positive correlation was also found between Cdk1 and cyclin B expression.

Bilateral asymptomatic common carotid artery stenosis: Mouse model for stroke research.

Background: Asymptomatic carotid artery stenosis has become more prevalent worldwide and is often associated with a poor prognosis. Numerous guidelines highlighted surgical interventions as treatment for carotid artery stenosis, but only a few recommendations were made regarding non-surgical interventions due to its limited data. Aims: This study aims to develop a mice model for research in non-surgical interventions of asymptomatic carotid artery stenosis. Methods: Adult male Rattus norvegicus, Wistar strain models with bilateral asymptomatic common carotid artery stenosis (BACAS) were created by ligating the common carotid artery with a 0.6 mm diameter needle and then removing the needle. The mice's body weight, clinical signs and symptoms, and post-mortem brain analysis were compared between the sham-operated group and the BACAS group. Results: The mortality rate among the BACAS group is 11.11%. There is no significant difference in mean body weight before surgery, after the observation period, and percentage of weight decrease between sham-operated and BACAS groups (p = 0.710, 0.632, and 0.806, respectively). None of the surviving mice in this study exhibit signs of motor paralysis. Gross examination of the brain reveals no signs of infarction or hemorrhage. Conclusion: We have established a novel BACAS mouse model which is cost-efficient, easy to produce, and with no significant alteration in body weight, clinical parameters, and brain morphology.

High Sucrose and Cholic Acid Diet Triggers PCOS-like Phenotype and Reduces Enterobacteriaceae Colonies in Female Wistar Rats.

Polycystic ovary syndrome (PCOS), a common hormonal disorder in women of reproductive age, is associated with a poor and unhealthy diet. This study aimed to investigate the effect of a high sucrose and cholic acid (HSCA) diet in the presence of PCOS-like phenotypes. Female Wistar rats were divided into HSCA and normal diet groups for four weeks, each with twenty rats. Body weight was assessed before and after the study. Blood and fecal samples were obtained to measure HOMA-IR and testosterone level (ELISA) and Enterobacteriaceae isolates grown on MacConkey Agar. Obtained ovarian tissues were H&E-stained. HSCA rats demonstrated a reduction in Enterobacteriaceae colonies (median 4.75 × 105 vs. 2.47 × 104/CFU, p < 0.001) and an elevated HOMA-IR (mean 2.94 ± 1.30 vs. 4.92 ± 0.51, p < 0.001), as well as an increase in testosterone level (median 0.65 vs. 3.00 ng/mL, p < 0.001), despite no statistical differences in the change in body weight (mean −2.31 ± 14.42 vs. −3.45 ± 9.32, p = 0.769). In H&E staining, HSCA rats had a reduction in preovulatory follicle count (median 0.50 vs. 0.00, p = 0.005). The HSCA diet caused insulin resistance and high testosterone levels, which contribute to the development of PCOS, and affected folliculogenesis by altering follicular maturation, but had no effect on ovulation.

Protective Effect of Apis dorsata Honey on Chronic Monosodium Glutamate-Induced Testicular Toxicity in Mus musculus Mice.

Objectives: This study proves the protective effect of Apis dorsata honey against chronic monosodium glutamate (MSG)-induced testicular toxicity on the Leydig cell necrosis count and malondialdehyde (MDA) serum level in Mus musculus mice. Materials and Methods: In this study, 25 male mice were used and grouped into two large groups: The control group consisting of negative control (C-) and positive control (C+). C+ group was fed with 4 mg/g body weight (gBW) of MSG followed by distilled water. The treatment group consisted treatment 1, treatment 2, and treatment 3 groups with A. dorsata honey dosage 53.82 mg/20 g, 107.64 mg/20 g, 161.46 mg/20 g per os (p.o.), respectively, followed by MSG 4 mg/g BW of MSG p.o. For the difference analysis between the group used the one-way ANOVA test and Duncan test. Results: The result of this study showed that there was a significant difference between the treatment group and control group (p<0.05) in the Leydig cell necrosis count and MDA levels. The highest Leydig cell necrosis count and MDA level were found in C+ with values 13.20 ± 2.05 cell and 37.08 ± 9.17 µmol/L compared to C-, while in the treatment group, T3 showed the lowest Leydig cell necrosis value and MDA level 4.64 ± 0.55 cell and 14.22 ± 2.01 µmol/L compared to the C+ group. Conclusion: It can be concluded that A. dorsata honey could reduce the Leydig cell necrosis number and MDA level of mice (Mus musculus) exposed to MSG.

Beneficial effects of kebar grass (Biophytum petersianum klotzsch) ethanol extract to increase motor reflex and spatial memory in mice offspring (Mus musculus) from lactating mothers exposed to carbofuran.

Background and purpose: This study aimed to determine the potency of kebar grass ethanol extract to overcome an increase in cerebellar neuronal cell necrosis, which has an impact on decreasing motor reflex function and spatial memory of mice from lactating mothers exposed to carbofuran. Experimental approach: Forty lactating mice were divided into four groups, 10 each; including control, T1 (carbofuran 0.0125 mg/day), T2 (vitamin C 5 mg + carbofuran 0.0125 mg/day), T3 (kebar grass extract 3.375 mg + carbofuran 0.0125 mg/day). The mice were orally administered with carbofuran, vitamin C, and kebar grass extract on days 0 to 14 postnatal. On the 15th day, brains of the mice were necropsied to measure the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH), H&E staining; motor reflex tests were performed on 10-day-old mice, and the mice aged 30 days were tested on their swimming and spatial memory. Findings / Results: Carbofuran caused an increase in MDA, GSH, neuronal cell necrosis, surface righting reflex, a decrease in SOD, swimming ability, and spatial memory. Kebar grass extract and vitamin C administration decreased MDA, GSH, neuron necrosis, surface righting reflex, and increased SOD, swimming ability, and spatial memory. Conclusion and implications: Exposing to carbofuran in lactating mice caused brain oxidative stress, impaired motor reflexes, and spatial memory in mice offspring. Kebar grass extract and vitamin C administration prevented brain oxidative stress and inhibited disorders in motor reflexes, and spatial memory in mice offspring. Kebar grass extract administration was more effective than vitamin C.

Periosteum-induced ossification effect in skull defect through interleukin-8 and NF-κB pathway: An experimental study with Oryctolagus cuniculus rabbits.

Background: The purpose of this study was to analyze the response of inflammatory cytokines interleukin-8 (IL-8) and NF-κB to the closure of skull defect with periosteum as a scaffolding material in bone healing used after surgery. Methods: Thirty Oryctolagus cuniculus rabbits underwent a craniotomy to create a 20 mm diameter round defect in the parietal bones. The parietal bones were returned to its place and stabilized by an internal plate fixation. The defects were either left empty or implanted with periosteum. At 6 weeks, the specimens were euthanized and examined. Results: Histological examination showed a more well-developed formation of woven bone in the periosteum group. Immunohistochemical examinations showed that the use of periosteum in the closure of skull defects reduced the NF-κB and IL-8 response which affected the ossification process. Conclusion: The experiment showed that the use of periosteum was linked with IL-8 and NF-κB downregulation toward ossification effects at any point throughout the trial. Periosteum usage might be beneficial as a scaffolding material in bone healing for autograft cranioplasty in animal model and could be applied to clinical practice.

Vitamin D inhibited endometriosis development in mice model through interleukin-17 modulation.

Background: Endometriosis is a common, benign, estrogen-dependent, and chronic gynecological disease. Immune system disturbance and inflammatory abnormalities were involved in the pathogenesis of endometriosis. Therefore, it is logical to use vitamin D, which has an immunomodulatory capacity, as supportive therapy for endometriosis. Aim: This research aimed to study the effect of different doses of vitamin D on Interleukin-17 (IL-17) expression in endometriosis mice models. Methods: Endometriosis was induced in 24 mice divided into 4 groups of 6. Group C received no treatment, while groups T1, T2, and T3 received graded doses of oral vitamin D, sequentially 8, 16, and 24 IU, for 3 weeks. IL-17 expression and the extent of endometriotic peritoneal lesions were then measured and analyzed. Statistical tests were performed to see the difference in the mean area of endometriosis lesions and IL-17 expression between the control and treatment groups, as well as the correlation between the extent of endometriosis lesions and IL-17. Results: Endometriosis lesions decreased after 16 and 24 IU of vitamin D administration (p 0.023 and 0.009). Endometriosis lesion also tends to be smaller after 8 IU of vitamin D supplementation, although insignificant (p > 0.05). IL-17 expression was significantly lower after 24 IU vitamin D administration compared to the untreated group (p = 0.004). Lower IL-17 expressions were also observed after 8 and 16 IU vitamin D administration, although insignificant (p = 0.452 and p = 0.645). IL-17 expression was moderately and positively correlated with the extent of endometriosis lesions (p = 0.012, rho = 0.505). Conclusion: By modulating the expression of IL-17 in endometriotic lesions, vitamin D inhibited the development of endometriotic lesions in the endometriosis mice model. The recommended vitamin D dose in this study was 24 IU.

Effects of bone marrow mesenchymal stem cell transplantation on tumor necrosis factor-alpha receptor 1 expression, granulosa cell apoptosis, and folliculogenesis repair in endometriosis mouse models.

BACKGROUND AND AIM: Endometriosis affects the ovaries and causes a decrease in the oocyte quality during endometrial receptivity. During the development of ovarian follicles, paracrine communication occurs between granulosa cells and oocytes. This study was conducted to determine the effects of bone marrow mesenchymal stem cell transplantation on tumor necrosis factor-alpha (TNF-α) receptor 1 (TNFR1) expression, granulosa cell apoptosis, and folliculogenesis in endometriosis mouse models. MATERIALS AND METHODS: This study involved 42 female mice, which were divided into three groups: Healthy mice (T0), endometriosis mice without transplantation (T1), and endometriosis mice with bone marrow mesenchymal stem cell transplantation (T2). The mice were injected intraperitoneally with endometrial fragments (200 µL) to become endometriosis models. On day 15, the endometriosis models received mesenchymal stem cells. Sample collection was performed on day 29. Granulosa cell apoptosis and TNFR1 expression were examined using immunohistochemical staining, and folliculogenesis was assessed using hematoxylin and eosin staining of ovary samples. The data obtained from both examinations were statistically analyzed using Statistical Package for the Social Sciences. RESULTS: The results showed that TNFR1 expression is significantly decreased in T2 (p<0.004). The apoptosis of granulosa cells was lower in T2 (p<0.000). The primary, secondary, and graafian follicle counts in T2 were significantly increased. CONCLUSION: Bone marrow mesenchymal stem cell transplantation in endometriosis mouse models can reduce TNFR1 expression and granulosa cell apoptosis and improve folliculogenesis.

Effect of kebar grass extract ( Biophytum petersianum Klotzsch) on histopathological changes in liver of mice offspring from the parent exposed to carbofuran during lactation period.

This study aims to determine the potential of Kebar grass extract in reducing the impact of liver damage in mice offspring ( Mus musculus) from parent exposed to carbofuran during lactation period. 42 lactation mice ( Mus musculus) used in the study were divided into seven groups, each group consisting of six mice. Carbofuran, Kebar grass extract, and vitamin C are administered orally on days 1 to 14 after birth. This group consisted of K (aquadest), P1 (carbofuran 1/4 LD50 0.0125 mg/day), P2 (carbofuran 1/8 LD50 0.00625 mg/day), P3 (Kebar grass extract 3.375 mg (0.2 ml) + carbofuran 1/4 LD50), P4 (Kebar grass extract 3.375 mg (0.2 ml) + carbofuran 1/8 LD50), P5 (vitamin C 5 mg (0.2 ml) + carbofuran 1/4 LD50), and P6 (vitamin C 5 mg (0.2 ml) + carbofuran 1/8 LD50). On the 15th day after birth, mice were sacrified and their liver taken for microscopic examination with hematoxilin and eosin staining. The scoring data were analyzed using Kruskal-Wallis and Mann-Whitney test. The result showed significant different (p⟨0.05) among the treatment groups. Mean of P4 in degeration is (1.13), necrosis (1.13) and inflamation (0.73), while the mean of P6 in degeneration is (2.20), necrosis (2.73) and inflamation (1.93). The conclusion of this research is giving Kebar grass extract is more effective in reducing degeneration, necrosis and inflammatory cell's infiltration than vitamin C in in mice offspring ( Mus musculus) from parent exposed to carbofuran during lactation period.

The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation.

BACKGROUND AND AIM: The combination of vitrification techniques and in vitro maturation can reduce oocyte competence. Mitogen-activated protein kinase and maturation-promoting factor are significant in oocyte meiotic maturation regulation. This study aimed to analyze vitrification's effect, after warming followed by in vitro maturation, on the expressions of protein 38 (p38), cyclin-dependent kinase 1 (CDK1), and cyclin B and oocyte maturation level. MATERIALS AND METHODS: Immature goat oocytes were soaked in vitrification and warming solutions. The procedure was followed by in vitro maturation and in vitro maturation without post-warming vitrification as a control. These oocytes, along with their cumulus, were vitrified using hemistraw in liquid nitrogen. Oocyte maturation was carried out in a maturation medium that was added with 10 µg/mL of FSH, 10 µg/mL of LH, and 1 µg/mL E2 for 22 h. The expressions of p38, CDK1, and cyclin B were observed using immunocytochemical methods, which were assessed semiquantitatively according to the modified Remmele method. The oocyte maturation level was observed using the aceto-orcein staining method based on the achievement of chromosomes up to the metaphase II stage and/or the formation of the polar body I. RESULTS: p38 expression in vitrified oocytes after warming, followed by in vitro maturation, increased insignificantly (p≥0.05), with the acquisition of 3.91±2.69 and 2.69±0.50 in the control oocytes. CDK1 expression in vitrified oocytes decreased significantly (p≤0.05) after warming, followed by in vitro maturation, with the acquisition of 2.73±1.24 and 7.27±4.39 in the control oocytes. Cyclin B expression in vitrified oocytes decreased insignificantly (p≥0.05) after warming, followed by in vitro maturation, with the acquisition of 3.09±1.4 and 4.18±2.61 in the control oocytes. The proportion of vitrified oocyte maturation levels after warming, followed by in vitro maturation, decreased significantly (p≤0.05), with the acquisition of 45.45% and 77.27% in the control oocytes. CONCLUSION: This study concluded that vitrification after warming resulted in an insignificant increase in p38 expression, a significant decrease in CDK1 expression, an insignificant decrease in cyclin B expression, and a significant reduction in oocyte maturation levels.

The profiling of pre- and post-warming DNA in mouse embryos with microsatellite method.

AIMS: This research aimed to identify the deoxyribonucleic acid (DNA) profile and changes of post-warming embryo after being frozen with vitrification method using microsatellite method. MATERIALS AND METHODS: This research examined the mouse embryo blastocysts that were divided into four groups: Post-warming living blastocyst, post-warming living blastocyst with half fragmented cell, post-warming dead blastocyst, and pre-freezing living blastocyst. The isolation sample applied phenol-chloroform method. After obtaining polymerase chain reaction results, all the samples of pre-freezing fresh embryo, post-warming living embryo, dead embryo, and degenerated embryo were then examined by single-strand conformation polymorphism (SSCP). RESULTS: The amplification with D18mit14 primer was 100 bp and 150bp with D18mit87 primer, 150bp with D7mit22, and 300bp with D7mit25. The result of SSCP with D18mit14 primer showed that the blastocysts were fragmented and dead after warming process and formed into two DNA strand fragments, while the fresh embryos which passed freezing process did not form any fragment. D18mit87 primer SSCP indicated different fragments for each treatment. The result of SSCP using D7mit22 formed two different fragments for each treatment. While using D7mit25, the SSCP result formed some different fragments for each sample. Post-warming living embryo had similar ribbon to pre-freezing fresh embryo. CONCLUSION: D7mit222, D7mit25, and D18mit87 primers could be used as the aneuploidy marker on mouse embryos that were induced by post-warming process. The profile of living blastocyst, dead blastocyst, and post-warming fragmented blastocyst had different DNA tapes.