[Posterior urethral polyp: a rare case]

Urethral polyps are a rare finding in children. Urethral fibroepithelial polyps are usually discovered in the first decade of life. They present with voiding dysfunction, obstructive and irritative urinary symptoms, and hematuria. They may be associated with other congenital urinary tract anomalies. Histopathologically, they are usually benign lesions with no tendency to recurrence and are treated by surgical ablation, fulguration or laser therapy. Diagnosis is made by sonography, voiding cystourethogram and cystoscopy. In this report, a rare case of posterior urethral polyp in an eight -year old male child is presented

[Three dimensional culturing of human jaw osteoblasts in PLLA/HA scaffold]

Tissue engineering using somatic cells and synthetic extracellular matrix [scaffold] represents a new approach for regeneration of mineralized tissue and bone. This study was carried out to investigate the ability of a PLLA/HA scaffold to culture osteoblast cells in a three dimensional milieu. Three bony samples were taken from extraction sites during surgical extraction of wisdom teeth. Osteoblasts were obtained from specimens by using trypsin and collagenase and were cultured in monolayer up to passage four. Cells were seeded on PLLA/HA scaffolds at density of 106 cells/ml and then incubated for 21 days. The seeded cells were evaluated by Hoechst, von kossa and H and E stainings and scanning electron microscopy. Cellular growth was more pronounced when isolation was carried out by collagenase. According to scanning electron microscopy, osteoblast cells had been proliferated and attached to the scaffolds. H and E, Hoechst and von kossa stainings confirmed the presence of the harvested cells in the scaffold too. Our findings suggest that osteoblast cells can grow onto PLLA/HA scaffold in vitro

In vitro survival rate of bovine oocytes following vitrification in glass capillary micropipette [GCM]

The purpose of this study was to evaluate the use of glass capillary micropipette [GCM] as a vessel for vitrification of bovine oocytes. Cumulus-oocyte complexes [COCs] were obtained from slaughter-house and washed 5 to 6 times in the washing medium [TCM-199 + 20% FBS] and randomly assigned to treatment and control group. In the first step of vitrification, COCs were exposed to first vitrification solution [VS1] [10% ethylene glycol [EG], 10% DMSO in holding medium [TCM-l99 + 10% FBS: HM]] for I min at room temperature and then placed in VS2 solution [20% EG, 20% DMSO in HM] for 25 sec and immediately were loaded into the GCM vessel. The filled portion of GCM vessels were placed in liquid nitrogen [LN [2]] for 3 to 5 sec and then completely immersed and stored there. The oocytes were thawed by immersing the capillary end of the straw in 1 ml of 0.25 M sucrose in HM and gently expelling the contents. After 1 min the oocytes were transferred into 100 micro l of 0.15 M sucrose in HM for another 5 mm and then washed with HM twice. For examining the in vitro developmental potential of vitrified-warmed oocytes, the oocytes were placed in 50 micro 1 droplet of maturation medium [TCM-l99 + 10% FBS - 10 IU/ml PMSG + 15 IU/ml HCG] covered with paraffin oil in a CO [2] incubator at 38.5°C for 24 hrs. A high proportion of morphologically normal oocytes [90%] was recovered after vitrification-warming. The percentage of live oocytes after 24 hrs when tested with trypan blue in GCM group was 85.18%, significantly did not differ from control group [90%]. The proportion of oocytes which were found to have undergone nuclear maturation did not show statistical difference between the control and GCM group [61.29% vs 40%, respectively]. The results of present study demonstrated that vitrification of immature bovine oocytes in the GCM vessels and EG + DMSO solution have high survival rate

[ laboratory study on human jaw osteoblastic stem cells culturing]

The goal of this study was to evaluate different methods of cultivating human bone cells. Five periosteal and bone specimens obtained from the human jaw were divided into small pieces in the laboratory. After the addition of trypsin and collagenase enzymes and releasing of cells, three primary periostal, endosteal, and bony chips cells were prepared. After passing the required time for the growth of specimens, lamella were prepared and stained with alkaline phosphatase [ALP] in order to determine ALP positive cells. Mean time of cellular growth was 23 days. Human bone cells have the capability of being cultured under special sterile laboratory conditions and three dimensional culturing of them can be used for reconstruction of maxillofacial region defects

[Effects of Morphine dependency on the ovarian folliculogenesis following superovulation in mice]

Opioids may affect hypothalamic GnRH secretion and Hypothalamic-Pituitary- Gonad axis, resulting in reproductive disturbances. Current study investigates the effects of morphine on structure of ovary following superovulalion through morphologic/morphometric studies. Twelve young female NMRI mice were allocated into treatment and control groups. Treatment group received oral morphine at final dose of 0.4mg/ml for 21 days. Physical dependency was proved by injection of naloxone [2mg/kg ip]. The mice were superovulated by 10 iu PMSG [ip] and 48 hours later were sacrificed by cervical dislocation. Ovaries were removed and H and E staining was done. Every 10[th] serial section, which represents nonrandom 10 percent sample was counted. Follicles were classified into small, growing, antral and atretic according to the diameter and number of follicular cell layers surrounding oocytes. The volume and the weight of ovaries were recorded. In addition, the diameter of the antral follicles and oocytes was carefully measured by a calibrated oculometer. The volume and the weight of ovaries showed no significant alterations in the two groups. The proportion of small and atretic follicles was statistically different in treatment and control groups [P<0.001]. According to our data, oral morphine did not alter the volume and the weight of the ovary. However, folliculogenesis was moderately affected by morphine and following superovulation the behavior of ovaries in the treatment group is comparable to the control group

[Effects of morphine dependency on the ovarian folliculogenesis following superovulation in mice]

Opioids may affect hypothalamic GnRH secretion and Hypothalamic-Pituitary- Gonad axis, resulting in reproductive disturbances. Current study investigates the effects of morphine on structure of ovary following superovulation through morphologic/morphometric studies. Twelve young female NMRI mice were allocated into treatment and control groups. Treatment group received oral morphine at final dose of 0.4mg/ml for 21 days. Physical dependency was proved by injection of naloxone [2mg/kg ip]. The mice were superovulated by 10 iu PMSG [ip] and 48 hours later were sacrificed by cervical dislocation. Ovaries were removed and H and E staining was done. Every 10[th] serial section, which represents nonrandom 10 percent sample was counted. Follicles were classified into small, growing, antral and atretic according to the diameter and number of follicular cell layers surrounding oocytes. The volume and the weight of ovaries were recorded. In addition, the diameter of the antral follicles and oocytes was carefully measured by a calibrated oculometer. The volume and the weight of ovaries showed no significant alterations in the two groups. The proportion of small and atretic follicles was statistically different in treatment and control groups [P<0.001]. According to our data, oral morphine did not alter the volume and the weight of the ovary. However, folliculogenesis was moderately affected by morphine and following superovulation the behavior of ovaries in the treatment group is comparable to the control group

Effects of L-arginine on development of mouse pre-implantation embryos in culture media with high level of glucose

It is accepted that maternal hyperglycemia causes delay in early embryonic development, spontaneous miscarriage and malformations. According to various studies, some of these problems occur in earlier stages of embryonic developmen especially pre-implantation stage. It seems that elevated glucose level of blood can have important role in this regard as potential teratogen factor. One of cases, which can be related to racousnesses resulting from glucose effects is Nitric Oxide [NO] system disorder in hyperglycemic condition. Some evidences show at first in hyperglycemic condition, L-arginine uptake of media by embryo increases and therefore leads to decrease amount of available L-arginine and since L-arginine is essential substrate for NO production, so it's decrease inhibits NO production. To examine this hypothesis, 2-cell embryos of mice were cultured in media of high concentration of glucose [30mM] and different concentrations of L-arginine [5, 10, 20 mM] and their growth and development were assessed and at the end, embryos were stained by Hoechst 33254 color and the number of their blactocysts were counted by use a Fluorescence microscope. Comparison of embryos culture in HTF culture media with different concentration of glucose and L-arginin showed in high glucose media up to 30 mM affects growth and development of embryos totally and decrease their blactocysts numbers, but addition of 5-10 mM L-arginine to this media significantly improves this condition. On the contrary addition of L-NAME [an antagonist of L-arginine] significantly inhibits the development of pre-implantation embryos. It seems that reduction in NO production in diabetes is due to decreases in amount of available L-arginine, because increase in L-arginine concentration in high glucose media up to 10 mM partially improves high glucose embryo toxicity. Base on acquired result, it seems use of L-arginine or material which cause NO release in media, can have important role in prevention of high glucosis embryo toxicity