Ovarian torsion in a 2-year-old girl: A case report.

Background: Ovarian torsion (adnexal torsion) is a rare event in pediatric patients which is primarily managed by pediatric general surgeons. Case presentation: This study presents a case of ovarian torsion in a 2-yr-old girl with a history of episodic lower abdominal pain, nausea, and vomiting for 2 days. Her physical examination was normal except for mild tenderness in the lower abdomen with no palpable mass. A color Doppler ultrasound was performed for further investigation, and an ovarian torsion was reported without sonographic signs of intussusception and acute appendicitis, so she underwent laparotomy. A relatively complete torsion was observed in the left ovarian pedicle. Initially, the left ovary and fallopian tube had a dark appearance, and 10-15% of the ovarian tissue was still normal. Detorsion of ovary was done and it was decided to preserve the ovary. After about 20 min, the color of ovary and fallopian tube returned to relatively normal, indicating normal blood flow. The patient was discharged 2 days later because a follow-up color Doppler ultrasound showed normal ovarian blood flow. Conclusion: The possibility of ovarian torsion must be considered in all female infants with suspicious abdominal pain.

Hydroalcoholic Extract of Ziziphus Jujuba Leaf to Prevent Ethylene Glycol and Ammonium Chloride-Induced Kidney Stones in Male Rat: Is it Effective?

PURPOSE: This study aimed to evaluate the effect of Ziziphus jujuba (Z. jujuba) leaf hydroalcoholic extract on the prevention/treatment of kidney stones. MATERIALS AND METHODS: Thirty-six male Wistar rats were randomly divided into six groups: control, Sham (kidney stone induction (KSI) by ethylene glycol 1% + ammonium chloride 0.25% through drinking water for 28 days), Prevention groups 1, 2 (KSI and Z. jujuba leaf (250 and 500 mg/kg, respectively) through gavage for 28 days), and Treatment groups 1, 2 (KSI and Z. jujuba leaf (250 and 500 mg/kg, respectively) from the 15th day). On the 29th day, the rats' 24-hour urine was assessed, the animals were weighed, and blood samples were taken. Finally, after nephrectomy and weighing the kidneys, tissue sections were prepared to examine the number of calcium oxalate crystals and tissue changes. RESULTS: The results indicated a significant increase in kidney weight and index, tissue changes, and the number of calcium oxalate crystals in the Sham group compared to the control; using Z. jujuba leaf considerably reduced them in experimental groups compared to the Sham. Body weight decreased in the Sham and experimental groups (except the prevention 2 group) compared to the control, while this observed reduction was lower in all experimental groups compared to the Sham. The mean urinary calcium, uric acid, creatinine, and serum creatinine in Sham and experimental groups (except the prevention 2 group) indicated a substantial increase compared to the control and decreased significantly in all experimental groups compared to the Sham. CONCLUSION: Hydroalcoholic extract of Z. jujuba leaf is effective in the reduction of calcium oxalate crystals forming, and its most effective dose was 500mg/kg.

In Vitro Implantation Model Using Human Endometrial SUSD2+ Mesenchymal Stem Cells and Myometrial Smooth Muscle Cells.

OBJECTIVE: This study evaluated a novel in vitro implantation model using human endometrial mesenchymal stem cells (EMSCs), SUSD2+, and myometrial smooth muscle cells (SMCs) that were co-cultured with mouse blastocysts as the surrogate embryo. MATERIALS AND METHODS: In this experimental study, SUSD2+ MSCs were isolated from human endometrial cell suspensions (ECS) at the fourth passage by magnetic-activated cell sorting. The ECS and SUSD2+ cells were separately co-cultured with human myometrial muscle cells for five days. After collection of mouse blastocysts, the embryos were placed on top of the co-cultured cells for 48 hours. The interaction between the embryo and the cultured cells was assessed morphologically at the histological and ultrastructural levels, and by expression profiles of genes related to implantation. RESULTS: Photomicrographs showed that trophoblastic cells grew around the embryonic cells and attached to theECS and SUSD2+ cells. Ultrastructural observations revealed pinopode and microvilli-like structures on the surfaces of both the ECS and SUSD2+ cells. Morphologically, the embryos developed to the egg-cylinder stage in both groups. Gene expression analysis showed no significant differences between the two groups in the presence of an embryo, but an increased expression of αV was detected in SUSD2+ cells compared to ECS cells in the absence of an embryo. CONCLUSION: This study showed that SUSD2+ cells co-cultured with SMCs could interact with mouse embryos. The co-cultured cells could potentially be used as an implantation model.

Morphological, Ultrastructural, and Molecular Aspects of In Vitro Mouse Embryo Implantation on Human Endometrial Mesenchymal Stromal Cells in The Presence of Steroid Hormones as An Implantation Model.

OBJECTIVE: This experimental study aimed to evaluate the effects of 17ß-estradiol (E2) and progesterone (P4) on the interaction between mouse embryo and human endometrial mesenchymal stromal cells, and gene expressions related to implantation [αV and ß3 integrins, interleukin-1 receptor (IL-1R), and leukemia inhibitory factor receptor (LIFR)] using an in vitro twodimensional model. MATERIALS AND METHODS: In this experimental study, the endometrial stromal cells were isolated enzymatically and mechanically, and cultured to the fourth passage. Next, their immunophenotype was confirmed by flow cytometric analysis as mesenchymal stromal cells. The cells were cultured as either the experimental group in the presence of E2 (0.3 nmol) and P4 (63.5 nmol) or control group without any hormone treatment. Mouse blastocysts were co-cultured with endometrial mesenchymal stromal cells in both groups for 48 hours. Their interaction was assessed under an inverted microscope and scanning electron microscopy (SEM). Expressions of αV and ß3 integrins, LIFR, and IL-1R genes were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Similar observations were seen in both groups by light microscopy and SEM. We observed the presence of pinopode-like structures and cell secretions on the apical surfaces of endometrial mesenchymal stromal cells in both groups. The trophoblastic cells expanded and interacted with the mesenchymal monolayer cells. At the molecular level, expression of IL-1R significantly increased in the hormonal treated group compared to the control (P≤0.05). Expressions of the other genes did not differ. CONCLUSION: This study has shown that co-culture of endometrial mesenchymal stromal cells with mouse embryo in media that contained E2 (0.3 nmol) and P4 (63.5 nmol) could effectively increase the expression of IL-1R, which is involved in embryo implantation. However, there were no significant effects on expressions of αV and ß3 integrins, LIFR, and on the morphology and ultrastructure of endometrial mesenchymal stromal cells.

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice.

BACKGROUND: Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. OBJECTIVE: The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. MATERIALS AND METHODS: Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin (0.2% w/v) for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham's F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology (Pap-staining) and viability (eosin-Y staining). Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD (sperm chromatin dispersion) and terminal deoxynucleotidyl transferase (TUNEL) assay. RESULTS: Following saccharin consumption, we had a reduction in sperm motility with respect to control animals (p=0.000). In addition, the sperm count diminished (17.70±1.11 in controls vs. 12.80±2.79 in case group, p=0.003) and the rate of sperm normal morphology decreased from 77.00±6.40 in control animals into 63.85±6.81 in saccharin-treated mice (p=0.001). Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one (p=0.001, p=0.002 respectively). CONCLUSION: Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice.

Zona pellucida birefringence and meiotic spindle visualisation of human oocytes are not influenced by IVM technology.

The aim of the present study was to investigate the relationship between the presence of the meiotic spindle and zona pellucida (ZP) birefringence with morphology of in vivo- and in vitro-matured human oocytes. Germinal vesicles (n=47) and MI (n=38) oocytes obtained from stimulated ovaries of patients undergoing intracytoplasmic sperm injection (ICSI) underwent IVM. Using a PolScope (OCTAX PolarAID; Octax, Herbon, Germany), the presence of spindles and ZP birefringence was assessed in both in vivo-matured (n=56) and IVM (n=56) oocytes. In addition, the morphology of each matured oocyte was evaluated microscopically. There were insignificant differences for ZP birefringence and meiotic spindle between the in vivo-matured and IVM MII oocytes. Subanalysis revealed that the rates of morphologically abnormal oocytes did not differ significantly between the two groups, except in the case of irregular shape (P=0.001), refractile body (P=0.001) and fragmented polar body (P=0.03), which were higher in IVM oocytes. In the case of in vivo-matured oocytes, a significantly higher percentage of oocytes with intracytoplasmic and both intra- and extracytoplasmic abnormalities have a low birefringent ZP (P=0.007 and P=0.02, respectively). There was no relationship between morphological abnormalities and spindle detection. The findings suggest that clinical IVM is a safe technology that maintains the high maturation rate and integrity of oocytes. In addition, the use of the non-invasive PolScope is recommended for the detection of oocytes most suitable for ICSI.

Effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice.

OBJECTIVE: Chronic ethanol abuse causes reproductive organ failure and infertility in both humans and laboratory animals. Since sperm has a critical role in reproductive function, the objective of this unique study was to evaluate the effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice. STUDY DESIGN: A total of 36 adult male mice were equally divided into four groups. Group 1 received ethanol (10%, v/v) containing saccharin (0.2%, w/v), group 2 received ethanol (5%, v/v) containing saccharin (0.1%, w/v), group 3 was treated with saccharin (0.2%, w/v) and group 4 served as control and fed on basal diet for 35 days. Finally, the left cauda epididymis of each animal was cut and placed in Ham's F10 medium. Retrieved spermatozoa were used to analyze count, motility, morphology and viability. Sperm chromatin condensation and DNA integrity were assessed by five different tests including chromomycin A3 (CMA3), toluidine blue (TB), sodium dodecyl sulfate (SDS), and SCD (sperm chromatin dispersion) and sperm apoptosis was assessed by TUNEL. RESULTS: Following ethanol consumption, the sperm count diminished in the ethanol-treated groups. A decrease in sperm motility and an increase in the rate of morphological abnormalities (coiled and broken tails) were seen in the experimental and saccharin groups in comparison with controls. We showed that ethanol consumption can disturb sperm DNA integrity and chromatin remodeling and it may also induce sperm apoptosis. The rates of sperm apoptosis were 51.57 ± 7.45 and 42.85 ± 6.76 in the high ethanol dose and low ethanol dose groups, respectively. CONCLUSION: The results showed that alcohol has negative effects on sperm parameters, chromatin/DNA integrity and apoptosis in mice. These alcohol-induced sperm anomalies may be dose-dependent.

Does women's age influence zona pellucida birefringence of metaphase ΙΙ oocytes in in-vitro maturation program?

BACKGROUND: In vitro maturation (IVM) is a promising treatment option for certain infertile women. Nowadays, with the aid of PolScope, it has become possible to evaluate zona pellucida (ZP) characteristics as a parameter of oocyte quality. Moreover, quality of oocytes can be influenced by many factors, such as patient's age. The PolScope system is a non-invasive technique to assess birefringent structures such as the meiotic spindle and ZP in living oocytes. OBJECTIVE: The aim was to determine the influence of the woman's age on ZP birefringence, a sign of oocyte quality, and morphology of in-vitro matured human oocytes using non-invasive polarized light (PolScope) microscopy. MATERIALS AND METHODS: ZP birefringence and morphology were determined in 105 retrieved oocytes from 58 women undergoing ICSI in two age groups (≥30 years and <30 years). The immature oocytes were selected and after IVM, the quality of metaphase ΙΙ (MII) oocytes was assessed. The oocytes abnormalities were classified as intracytoplasmic and extracytoplasmic abnormalities. RESULTS: Oocyte maturation rates were significantly reduced in ≥30 year's women (56%) in comparison with other age group (80.7%). In addition, the ZP birefringence was significantly higher in MII oocytes in the younger group compared with the older group (76.2% vs. 38.1%; p=0.00). Following morphologic assessment, the rates of oocytes with extracytoplasmic (p=0.02) and both abnormalities (extra- and intracytoplasmic) (p=0.01) were higher in aged versus the younger women. CONCLUSION: There was a positive relationship between advanced maternal age with decreased ZP birefringence and oocyte morphological quality in in-vitro matured human oocytes.