Minimally invasive management of juvenile cystic adenomyoma: report of three cases.

Background: Juvenile cystic adenomyosis (JCA) represents a rare form of focal adenomyosis in young women. Objectives: To determine safety and effectiveness of minimally invasive surgery (MIS for JCA). Materials and Methods: Three patients aged 16-30 years old presented with chronic pelvic pain [2016 - 2019]. Hormonal treatment failed in two cases. Cystic lesions in the myometrium (n=2), and the broad ligament (n=1) was detected on transvaginal 2D ultrasound (TV 2D US) and/or magnetic resonance imaging (MRI). The cyst was separate from the endometrium in all the cases, within the myometrium in two patients and in the right broad ligament in one case. The cystic lesions were confirmed on laparoscopy; and laparoscopic excision of the cysts with adequate repair of the myometrial beds were performed in all cases with fertility preservation. Robotic assistance was chosen in one case in an attempt to avoid injury of the fallopian tube based on the cyst location during a previous laparoscopy. The endometrial cavity was entered in one case. Main outcome measures: Absence of intraoperative complications and relief of presenting symptoms postoperatively. Results: Pathology report confirmed the diagnosis of JCA is all cases. There were no intraoperative complications. All three patients reported relief of their symptoms 6 to 8 months after surgery. No recurrence of the JCA was reported using TV 2D US in all cases. Conclusions: MIS could be the treatment of choice for patients with JCA. The technique described in our study is safe, effective, and easy to master in experienced hands. What is new?: Value of MIS in treatment of patients with JCA.

Postnatal management of antenatally diagnosed ureteropelvic junction obstruction.

OBJECTIVE: The management of antenatally diagnosed ureteropelvic junction obstruction (PUJO) is controversial. Here, we present our experience over a period of 13 years and discuss our management protocol. MATERIALS AND METHODS: We reviewed the files of 234 patients with antenatally diagnosed congenital hydronephrosis due to PUJO. Management was tailored to each patient, based on a combination of diethylenetetraminepentacetic acid renogram outcome, pelvic diameter, as well as the patient's symptoms. RESULTS: Pyeloplasty was carried out early in 52 kidneys. Of the total, 182 patients were managed expectantly. Out of these, 45 underwent delayed pyeloplasty. The remaining 137 patients (189 units) were managed non-operatively. In patients who underwent early pyeloplasty, the mean split differential renal function was 37% before and 40.05% after surgery. In those with delayed intervention, the mean renal function was 37.8% before and 42.2% after surgery. In patients who did not have surgery, the mean differential renal function was 45.7% initially and 48.2% at the last accepted follow up. The overall operative success rate was 97.9%. CONCLUSION: We believe that pyeloplasty is the proper treatment for babies with congenital PUJO and <40% split differential function and/or pelvic diameter >35 mm at the initial visit. In other patients a period of observation is warranted, and pyeloplasty should be carried out only if their kidney function deteriorates or the renogram curve does not show improvement. With the excellent results of pyeloplasty we believe that a safer approach is to operate on more kidneys rather than risk of losing valuable kidney function.

Laparoscopic removal of an autoamputated ovarian cyst in an infant.

Ovarian cysts autoamputation is an extremely rare complication. All reported cases were removed by laparotomy. A successful laparoscopic removal is presented.

Transformation of protoplasts and intact cells from slowly growing embryogenic callus of wheat (Triticum aestivum L.).

A procedure for culturing protoplasts from slowly growing embryogenic calli of wheat was developed. The procedure was dependent on the ability to isolate large numbers of culturable protoplasts from slowly growing embryogenic callus. Approximately 68% of the isolated protoplasts divided, and 22% formed colonies; of the latter, 67% continued to proliferate. Plating efficiency was reduced when protoplasts were transformed by polythylene glycol, electroporation, and/or Agrobacterium. Intact cells were also directly transformed by electroporation. Direct electroporation of the Agrobacterium binary vector into intact cells resulted in a significant increase of GUS activity over the control.

Factors affecting transient gene expression in protoplasts isolated from very slowly growing embryogenic callus cultures of wheat (Triticum aestivum L.).

Protoplasts isolated from embryogenic ('Mustang' and 'Chinese Spring') and non-embryogenic ('Mit') calli of wheat (Triticum aestivum L.) genotypes transiently expressed ß-glucuronidase (GUS) activity when electroporated with a plasmid containing the GUS gene and driven by an enhanced 35S promoter and a TMV leader sequence. Conditions for the maximum expression of GUS activity were: electroporation of the freshly isolated protoplasts at 250 Vcm(-1) and 250 µF for 2 s using 50 µg/ml of plasmid DNA; incubation of the protoplasts with the plasmid before the pulse for 2 h; and a 15-min recovery period on ice after the pulse. In general, a higher GUS activity was obtained in protoplasts of non-embryogenic (NE) callus origin than in those of embryogenic (E) callus origin. Only GUS constructs containing a duplicate 35S promoter derivative resulted in a significant level of GUS expression. The presence of the TMV viral leader sequence in the pAGUS1-TN2 plasmid construct resulted in a significant increase of GUS activity in the electroporated protoplasts of both callus types. On the other hand, protoplasts electroporated with the Adh1 promoter and intron showed a threefold less GUS activity than those electroporated with pAGUS1-TN2. Optimized conditions for DNA uptake and expression were very similar for protoplasts of both callus types. The importance of these findings for the successful regeneration of transgenic and fertile wheat plants is discussed.

Factors controlling variability in measurements of the transient gene expression in electroporated protoplasts of wheat callus.

Factors controlling variability in enzyme transient expression assays have been investigated in electroporated protoplasts isolated from wheat embryogenic and nonembryogenic calli. The level of variation was reduced to a minimum through the optimization of the beta-glucuronidase measurements in the pellet and the supernatant of the homogenized protoplasts, by expressing the data on the basis of the number of protoplasts found to be viable immediately before the assay and on the amount of protein in the pellet and supernatant. Protoplast separation on the basis of size was also useful in eliminating some of the variation resulting from the heterogeneity of the callus used. Efforts to partially synchronize the callus tissue by auxin starvation have not resulted in a significant decrease of this variation. Our results indicate that the level of variation in enzyme transient activity in protoplasts resulting from calli can be reduced by implementation of the experimental techniques presented here.

Restoration of regeneration potential of long-term cultures of red fescue (Festuca rubra L.) by elevated sucrose levels.

A tissue culture protocol for restoring embryogenic ability and increasing green plant regeneration from long-term callus (5-year old) and suspension cultures of "Dawson" red fescue (Festuca rubra var trichyoplylla Gaud) was developed. Pretreatment with elevated levels of sucrose over the standard level (60 mM) enhanced regeneration capacity and decreased the number of albino plants. The highest degree of embryogenesis and green shoot number occurred when calli were pre-treated on MS basal medium supplemented with 120 mM sucrose. Mannitol caused callus discoloration and death if added to pre-treatment media at 60, 90, 120, 150 or 180 mM. Cell suspension growth was greatest when 135 mM sucrose was added to the pre-treatment growth media. High concentrations of sucrose (135 and 180 mM) were necessary for plant regeneration from suspension aggregates pretreated with 135 or 180 mM sucrose and then plated on a growth regulator-free regeneration medium composed of half-strength MS salts and B5 vitamins.

Isolation and culture of protoplasts from embryogenic suspension cultures of red fescue (Festuca rubva L.).

Protoplasts were isolated from fast-growing embryogenic suspension cultures of red fescue cv. Dawson (Festuca rubra L.) without agitation. The enzyme isolation solution was highly efficient at releasing protoplasts of greater than 95% viability (5×10(6)-10(7) protoplasts per ml of packed cell volume). A three step procedure was followed for washing and transferring protoplasts from a solution high in inorganic salts to a medium containing glucose and sucrose. The addition of 30 mM sodium thiosulfate to the wash and culture media was found to be helpful in reducing the number of lysed protoplasts. Isolated protoplasts began to divide within 48-72 h when protoplasts were plated in agarose squares and surrounded by nurse cells (mixed nurse plating technique). Maximum colony formation (plating efficiency) was approximately 1%. Many of the colonies continued to grow and produced embryos when transferred to a medium consisting of half-strength MS salts, 4 mg/l 2,4-D, 3 g/l casein hydrolysate and 30 g/l sucrose. Upon transfer to hormone-free medium and exposure to light 16 h/day, many of the embryos germinated to produce green leaves and roots.