Liquid Chromatography-Mass Spectrometry-Based In Vitro Metabolic Profiling Reveals Altered Enzyme Expressions in Eicosanoid Metabolism

BACKGROUND: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. METHODS: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. RESULTS: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R 2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. CONCLUSIONS: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.

Effect of Modulation of hnRNP L Levels on the Decay of bcl-2 mRNA in MCF-7 Cells

It has been shown that CA repeats in the 3'-untranslated region (UTR) of bcl-2 mRNA contribute the constitutive decay of bcl-2 mRNA and that hnRNP L (heterogenous nuclear ribonucleoprotein L) interacts with CA repeats in the 3'-UTR of bcl-2 mRNA, both in vitro and in vivo. The aim of this study was to determine whether the alteration of hnRNP L affects the stability of bcl-2 mRNA in vivo. Human breast carcinoma MCF-7 cells were transfected with hnRNP L-specific shRNA or hnRNP L-expressing vector to decrease or increase hnRNP L levels, respectively, followed by an actinomycin D chase. An RT-PCR analysis showed that the rate of degradation of endogenous bcl-2 mRNA was not affected by the decrease or increase in the hnRNP L levels. Furthermore, during apoptosis or autophagy, in which bcl-2 expression has been reported to decrease, no difference in the degradation of bcl-2 mRNA was observed between control and hnRNP L-knock down MCF-7 Cells. On the other hand, the levels of AUF-1 and nucleolin, transacting factors for ARE in the 3'UTR of bcl-2 mRNA, were not significantly affected by the decrease in hnRNP L, suggesting that a disturbance in the quantitative balance between these transacting factors is not likely to interfere with the effect of hnRNP L. Collectively, the findings indicate that the decay of bcl-2 mRNA does not appear to be directly controlled by hnRNP L in vivo.

The Clinical Effects of Amitriptyline in Patients with Urinary Frequency and Chronic Pelvic Pain Syndrome / 대한비뇨기과학회지

PURPOSE: We evaluated the effects of amitriptyline in patients with urinary frequency or pelvic pain syndrome and compared with propiverine hydrochloride (BUP-4(R)) and diazepam (Valium(R)). MATERIALS AND METHODS: One-hundred and sixty-eight patients with urinary frequency or pain were included and 38 out of 168 patients were lost to follow up. The patients with interstitial cystitis, UTI and neurogenic bladder were excluded. Amitriptyline group were instructed to take 25mg to 75mg of amitriptyline before bed. Propiverine hy drochloride and diazepam group were instructed to take 20mg, 4-6mg before bed, respectively. The clinical effects and side effects were evaluated after 4 weeks of medication. RESULTS: In the amitriptyline group, 73.1% in 78 patients with frequency, 68.3% in 60 pain, 65.6% in 32 nocturia, 53.8% in 26 weak urinary stream, 53.8% in 13 dysuria, 72.7% in 11 urgency showed symptom improvement. Urinary frequency and pain score were significantly decreased in the amitriptyline group. In the propiverine hydrochloride group, 66.7% in 21 patients with frequency, 38.8% in 18 pain showed symptom improvement. In the diazepam group, 37.5% in 24 patients with frequency, 31.3% in 16 pain showed symptom improvement. Side effects of amitriptyline had appeared in 24 (27.6%) out of 87 patients; 17 cases of dry mouth, 10 drowsiness, 2 agitation, 2 nausea, 1 constipation and 1 skin rash. CONCLUSIONS: Amitriptyline is useful in vague voiding symptoms especially in urinary frequency and chronic pelvic pain syndrome. It is equally effective in both men and women. Side effects are minimal and tolerable. A double-blind placebo-controlled trial is necessary for general acceptance in the future.

Impalpable Testes: An Experience of 97 Surgically Corrected Cases / 대한비뇨기과학회지

PURPOSE: We analyzed our experience of surgically corrected impalpable testes to evaluate our results and to determine the best possible surgical approach for impalpable testes. MATERIALS AND METHODS: Medical records of 97 boys who had undergone surgery for impalpable testes were reviewed with respect to the age at presentation, localization procedure, surgical approach, testicular location and volume. Minimal follow-up was 6 months. RESULTS: The average age at presentation was 47.7 months. Surgical locations of testes included preperitoneum in 46 testes, intraabdomen in 18 testes and inguinal canal in 1 testis. Fifty-three testes were absent at exploration. Surgical procedures consisted of 59 orchiopexies, 53 excision of blind-ended spermatic vessel and 6 orchiectomies. The average volume of abdominal testes was reduced to 44.0% of contralateral normally descended testicular volume. Compensatory hypertrophies were seen in 45 contralateral testes among 53 unilateral absent testes. The inguinal exploration was successful in defining the location of all the testes in the last 41 consecutive boys. Laparoscopy did not help to avoid surgical exploration. Of 38 impalpable testes at office examination, 18 testes turned out to be palpable when examined under anesthesia while ultrasonography detected 14 testes with 1 false positive finding. Of 51 regular orchiopexies 49 testes showed excellent or acceptable results. Of 8 Fowler-Stephens orchiopexies 3 testes resulted in atrophies. CONCLUSIONS: Most pexed impalpable testes showed successful results in terms of testicular position and growth. However, there were more testicular atrophies in cases whose spermatic vessels were ligated. Inguinal exploration with or without intraperitoneal extension was successful in the management of impalpable testes in this series.

A Case of Bilateral Renal Angiomyolipoma with Lymph Node Involvement / 대한비뇨기과학회지

Renal angiomyolipomas are relatively common benign neoplasms of kidney, composed of mature fat cells, thick-walled blood vessels and smooth muscle. There are two different types: 1) the solitary angiomyolipoma without other disease, 2) bilateral or fragmentary multiple angiomyolipoma in tuberous sclerosis. Regional lymph nodes can be rarely involved in renal angiomyolipoma, that may not represent malignant spread but reflect the multicentricity. We report a case of multiple bilateral renal angiomyolipoma without tuberous sclerosis, which involved regional lymph nodes simultaneously.

Bladder Exstrophy with Successful Initial Closure / 대한비뇨기과학회지

Bladder exstrophy is a rare condition with an incidence of between 1 in 10,000 and 1 in 50,000 live births and the male-to-female ratio ranges 2.3:1 to 6:1. The basic defect of bladder exstrophy is an abnormal overdevelopment of the cloacal membrane, preventing medial migration of the mesenchymal tissue and proper lower abdominal wall development. Although there are some recent successful reports of the one-stage repair of bladder exstrophy that involves complete primary repair of exstrophy and total disassembly of the epispadiac penis, staged repair still represents the most common operation for correction of this anomaly. A successful initial closure of the infant born with bladder exstrophy is the single most important determinant of continence in the staged repair of exstrophy patient. We report a case of bladder exstrophy with successful initial closure of bladder using posterior iliac osteotomy and paraexstrophy skin flap.