Cyclosporine A induces endothelin-converting enzyme-1: Studies in vivo and in vitro.

AIM: Cyclosporine A (CsA) induces renal vasoconstriction and hypoxia and enhances the expression of endothelin-1 (ET-1) pro-hormone (pre-pro-ET-1), plausibly leading to a feed-forward loop of renal vasoconstriction, hypoxia and enhanced synthesis of the potent vasoconstrictor ET-1. Endothelin-converting enzyme (ECE)-1 cleaves big endothelin to generate endothelin (ET)-1 and is upregulated by hypoxia via hypoxia-inducible factor (HIF). We hypothesized that in addition to the direct induction of ET-1 synthesis, CsA might also intensify renal ECE-1 expression, thus contributing to enhanced ET-1 synthesis following CsA. METHODS: CsA was administered to Sprague Dawley rats (120 mg/kg/SC) for 4 days, and renal HIF and ECE-1 expression were assessed with Western blots and immunostaining. Human umbilical vein endothelial cells (HUVEC) and proximal tubular cell line (HK-2) were subjected to CsA, and ECE-1 induction was evaluated using real-time mRNA PCR and Western blots. RESULTS: Cyclosporine A intensified renal parenchymal ECE-1 expression in the rat kidney, particularly in distal nephron segments, along with renal hypoxia (detected by pimonidazole adducts) and HIF expression, in line with our recent observations showing episodic hypoxia in mice subjected to CsA. Furthermore, in cultured normoxic HUVEC and HK-2 cells, CsA dose-dependently induced both pre-pro-ET-1 and ECE-1 mRNA and protein expression, with enhanced ET-1 generation. CONCLUSION: CsA induces ECE-1 via both hypoxic and non-hypoxic pathways. ECE-1 may contribute to increased renal ET-1 generation following CsA, participating in a feed-forward loop of renal parenchymal hypoxia and ET synthesis.

Crystallization Behavior of Amorphous Si3N4 and Particle Size Control of the Crystallized α-Si3N4.

Amorphous silicon nitride powder prepared by low-temperature vapor-phase reaction was heat treated at various temperatures for different periods of time to examine the crystallization behavior. The effects of the heat-treatment temperature and duration on the degree of crystallization were investigated along with the effect of the heat-up rate on the particle size, and its distribution, of the crystallized α-phase silicon nitride powder. A phase transition from amorphous to α-phase occurred at a temperature above 1400 degrees C. The crystallization. process was completed after heat treatment at 1500 degrees C for 3 h or at 1550 degrees C for 1 h. The crystallization process starts at the surface of the amorphous particle: while the outer regions of the particle become crystalline, the inner part remains amorphous. The re-arrangement of the Si and N atoms on the surface of the amorphous particle leads to the formation of hexagonal crystals that are separated from the host amorphous particle. The particle size and size distribution can be controlled by varying the heat-treatment profile (namely, the heat-treatment temperature, heating rate, and heating duration at the specified temperature), which can be used to control the relative extent of the nucleation and growth. The completion of most of the nucleation process by lowering the heat-up rate can be used to achieve a singlet particle size distribution. Bimodal particle size distribution can be achieved by fast heat-up during the crystallization process.

Uterine prolapse in a primigravid woman

Uterine prolapse during pregnancy is an uncommon condition. It can cause preterm labor, spontaneous abortion, fetal demise, maternal urinary complication, maternal sepsis and death. We report the case of uterine prolapse in a 32-year-old healthy primigravid woman. She had no risk factors associated with uterine prolapse. She was conservatively treated, resulting in a successful vaginal delivery. This report is a very rare case of uterine prolapse in a young healthy primigravid woman, resulting in a successful vaginal delivery.

Analysis of migrated hydrogel used for breast augmentation revealed prolamin (a cereal seed storage protein).

BACKGROUND: We report on a case of leakage and migration to the upper abdomen of an unknown injected material that was used for breast augmentation. It was revealed to be prolamin by Fourier transform infrared (FTIR) analysis and pyrolysis gas chromatography/mass spectrometry (PY-GC/MS). METHODS: A 35-year-old woman who had undergone mammary augmentation by transaxillary injection 8 years previously presented with a decreased size of her left breast and a palpable mass in the left upper quadrant (LUQ). Mammogram and ultrasonography showed multiple dense masses and several hypoechoic areas, respectively. Abdominal ultrasound showed a hypoechoic lesion between the subcutaneous layer and the abdominal wall muscles. When the left breast and the lump in the LUQ were explored, 90 and 160 cc of yellow, sticky, granular gel gushed out. FTIR analysis and PY-GC/MS were used to investigate the component of the removed gel. RESULTS: When this gel was analyzed by FTIR with the transmittance mode, intensity bands appeared at 3295.2 (NH2), 2927.2 (CH), 1650 (C=C), 1544.6 (C-C), and 1403.1 (C-N) cm(-1). The result showed a 93.84% match with purified zein, a 91.19% match with zein from corn, and a 90.27% match with poly(N-methyl acrylamide). FTIR with the attenuated total reflectance (ATR) mode revealed that the gel matched with wheat gluten flour. Based on the result of PY-GC/MS, the gel was suspected to be protein. CONCLUSION: This is the first such report on performing chemical analysis of a leaked injected gel from human breast implantation. The removed gel from the breast augmentation was revealed to be prolamin, which is a cereal seed storage protein. We think FTIR might be a useful tool for analyzing and confirming extracted materials that were previously injected to the body.

[Method of radiocapacity factor assessment in study of cross adaptations of plants].

Adaptation of plants to the action of gamma-irradiation and cadmium chloride has been studied in the experiments on water culture of maize plants. A new method of registration of the biological object state at the conditions of the stress-factor action - "the method of radiocapacity factor" based on the measurement of the culture medium radio-activity during the incubation process of vegetative objects has been developed. The method offered has allowed us to estimate in dynamics an integrated condition of the root system exposed to stimulating and inhibiting doses of acute gamma-irradiation. Application of the pattern of radio-adaptive response has allowed us to reveal it using both a conventional technique - by registering the values of growing parameters, and the radiocapacity factor method.

A Case of Staphylococcal Tricuspid Valve Endocarditis With Para-Aortic Abscess in a Patient With Bicuspid Aortic Valve

Paravalvular abscess is a serious complication of infective endocarditis. The aortic valve and its adjacent ring are more susceptible to abscess formation and paravalvular extension than the mitral valve. A 15-years old patient with bicuspid aortic valve presented with staphylococcal tricuspid valve endocarditis complicated by para-aortic abscess that ruptured into the aortic sinus. We report the clinical, laboratory and echocardiographic features and treatment of this patient and conduct a literature review on this subject.

A Case of Gastrointestinal Stromal Tumor of the Rectum Presenting as Constipation / 이화의대지

A gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the gastrointestinal tract and expresses CD117, a c-kit proto-oncogene, which can be detected immunohistochemically. We reported a GIST of the rectum of a 61-year-old-woman who had visited emergency room complaining of constipation over one week. Upon rectal examination, a round hard mass was palpated. Colonoscopy showed a 7x5 cm sized protruded lesion with surface ulceration on a rectum, adjacent the anus. And abdomen computed tomography revealed the soft tissue mass compressing anterolateral wall of the rectum and these findings suggest possibility of rectal submucosal tumor such as GIST. The patient had been treated with a ultra anorectal anastomosis with loop ileostomy. Immunohistochemical studies on the surgically resected specimen showed c-kit (+) and CD34 (+). The final diagnosis was a GIST of the rectum. She was grouped into high risk and she has been given adjuvant chemotherapy with Imatinib.

[Diagnosis of pulmonary heart in patients with chronic obstructive pulmonary disease].

Informative value of clinical, electrocardiographic, and echocardiographic diagnostic criteria of compensated chronic pulmonary heart (CPH) in patients with chronic obstructive pulmonary disease (COPD) was considered. The study included 229 patients with COPD of which 105 (group 1) showed no signs of CPH, 71 (group 2) had compensated and 53 (group 3) uncompensated CPH. They were examined by the standard echoCG method using an Acuson-128 HR apparatus (USA) and 12-lead ECG during a 2 year-long follow-up period. Direct cardiac clinical and electrocardiographic signs of right ventricular eccentric hypertrophy were found to have high informative value (100%) but very low sensitivity (7-53%). Indirect diagnostic criteria of compensated CHP are such non-specific signs as age of COPD patients above 50 yr, duration of the disease over 8 yr and broncho-obstructive syndrome (episodes of low-productive cough, dyspnea under small physical load and at rest, X-ray signs of lung emphysema, substantial reduction of FEV and FEV/FVC ratio) in 73-94% of the patients were associated with compensated CPH. The most valuable (75.8-90.5%) cardiographic diagnostic criteria for compensated CPH were MPAP > 22 mmHg (at rest), LVEDD > 24 mm, LVEDS > 17 mm, RVWT = > 5 mm, RAEDD > 32 mm. Criterion LVEDD > 24 mm has the optimal ratio of sensitivity (94.4%), specificity (85.7%), and positive predictive value (86.6%). Comprehensive clinical assessment of COPD character and duration, patients' age, manifestation of broncho-obstructive syndrome and direct clinical signs suggesting involvement of the right half of the heart permits to predict CPH in patients with COPD with a probability of 75%. The definitive diagnosis is verified by echoCG or other instrumental methods.

Diabetes and radiocontrast media increase endothelin converting enzyme-1 in the kidney.

Plasma endothelin-1 levels rise in diabetes and after exposure to contrast media suggesting a role in progressive diabetic and acute radiocontrast nephropathies. Here we studied individual and combined effects of streptozotocin-induced diabetes and contrast media on renal endothelin converting enzyme-1 levels in the rat. In vivo, medullary (but not cortical) endothelin converting enzyme protein gradually increased 4 to 5-fold following the induction of diabetes or after the administration of contrast media but rose 15-fold when diabetic rats were given contrast media. Changes in mRNA expression paralleled those of the protein. Immunohistochemistry confirmed that increased tubular and endothelial cell endothelin converting enzyme-1 were most pronounced in the medulla. In vitro, endothelin-1 levels increased 3-fold following incubation of endothelial cells with media high in glucose or with contrast and 4-fold with their combination. Endothelin converting enzyme-1 protein and mRNA expression changed in a similar pattern while prepro endothelin-1 mRNA increased with each insult but not in an additive way. Our study shows that diabetes and contrast media up-regulate renal medullary endothelin converting enzyme-1 expression and synthesis.

Adaptation to hypoxia in the diabetic rat kidney.

Hypoxia of the kidney in diabetes could predispose it to develop acute and chronic renal failure. To examine the relationship between renal hypoxia and renal failure, we measured hypoxia (as a pimonidazole adducts), hypoxia-inducible factors (HIFs), and a hypoxia target gene heme oxygenase-1. The studies were performed in rats with streptozotocin (STZ)-induced diabetes, Cohen diabetes sensitive rats, and during short-term artificial hyperglycemia in rats induced by intravenous glucose and octreotide. STZ-treated rats received insulin, the superoxide dismutase mimetic tempol, or contrast medium. Radiocontrast media causes hypoxia and HIF induction. Hypoxia, HIFs, and heme oxygenase were undetectable in controls, but transiently activated in STZ-treated and the Cohen diabetes sensitive rats. Different patterns of HIFs and pimonidazole were observed between the three models. Insulin abolished pimonidazole and HIF induction, whereas tempol lead to increased HIFs and heme oxygenase induction at similar levels of pimonidazole. When compared with control rats, STZ-treated rats exhibited more intense and protracted renal pimonidazole, with augmented hypoxia inducible factor production and reduced GFR following contrast media. Our data suggest that both regional hypoxia and hypoxia adaptation transiently occur in early stages of experimental diabetes, largely dependent on hyperglycemia or after contrast media. Tempol may augment the HIF response in diabetes.

[Monitoring of infectious diseases among wild boars].

Sixty-seven serum samples and 43 pathological material samples from wild boars, taken in 5 regions of Russia and in the Kharkov Region of the Ukraine in 2002 to 2005 were studied. Wild boars in some regions of Russia were shown to be carriers of Aujeszky's disease virus, porcine parvovirus, porcine circovirus type 2, lymphotropic herpesvirus-1, porcine cytomegalovirus, Mycoplasma hyopneumoniae and Pasteurella multocida. The classical swine fever (CSF) virus genome was detected by polymerase chain reaction in the samples from 10 wild boars from 2 Russian regions (the Tver and Moscow Regions). Sequencing of the E2 gene 5'-terminal region of detected CSF virus isolates showed that they were closely related to two field virus isolates early found in domestic pigs in the Moscow and Vladimir Regions, which suggests that there is an epizootic relation between the SCF outbreaks among wild boars and domestic pigs in these regions. Tests for porcine reproductive and respiratory syndrome, transmissible porcine gastroenteritis, porcine influenza, enteroviruses, and actinobacillus-induced pleuropneumonia were negative in all the regions under study.

Hypoxia-inducible factors and tubular cell survival in isolated perfused kidneys.

Adaptation to hypoxic environment is conferred through hypoxia-inducible transcription factors (HIFs). We have previously shown that the HIF system is transiently activated in vivo in radiocontrast-induced acute renal failure, associated with profound hypoxia in the renal medulla. Medullary thick ascending limbs (mTALs), the most affected nephron segments in this model, were virtually unable to mount an adaptive HIF response. Here, we study correlations between oxygenation, HIF activation, and cell viability in a related ex vivo model, the isolated perfused rat kidney (IPK). In IPKs perfused with cell-free oxygenated medium, severe medullary hypoxic damage developed, affecting 42+/-9% of mTALs in the mid-inner stripe. HIF-1alpha tubular immunostaining was noted with a zonal and tubular pattern largely similar to our findings in vivo: in 34+/-3% of collecting ducts (CDs) within the mid-inner stripe and extensively in the papillary tip, whereas mTALs were all HIF-negative. In IPKs supplemented with RBCs (improved oxygen supply), mTAL damage was totally prevented and CDs' HIF expression was attenuated (22+/-4%). By contrast, although measures designed to reduce medullary hypoxia by decreasing tubular reabsorptive activity (furosemide, ouabain, or high-albumin-non-filtering system) reduced mTAL damage, all paradoxically resulted in increased HIF expression in CDs (51+/-4%), and 17+/-3% of mTALs became immunostained as well. Our data confirm that CDs and mTALs have markedly different HIF responses, which correlate with their viability under hypoxic stress. mTALs transcriptional adaptation occurs within a narrow hypoxic range, and it appears that workload reduction can shift mTALs into this window of opportunity for HIF activation and survival.

[Recombinant non-structural 3A, 3B and 3AB proteins of foot-and-mouth disease virus: use for differentiation of vaccinated and infected cattle].

Recombinant proteins 3A, 3B and 3AB were obtained by expression in Escherichia coli and purified by metal-chelate chromatography. The proteins were used as antigens in indirect ELISA to differentiate vaccinated and infected cattle. While testing 200 sera from cattle 3A-ELISA was more sensitive and specific than 3B- and 3AB-ELISA. Compared with "Chekit FMD-3ABC", 3A-ELISA showed the same level of specificity and higher level of sensitivity.

Use of the recombinant nonstructural 3A, 3B, and 3AB proteins of foot-and-mouth disease virus in indirect ELISA for differentiation of vaccinated and infected cattle.

Recombinant foot-and-mouth disease virus (FMDV) proteins 3A, 3B, and 3AB were produced by expressing the corresponding genes in Escherichia coli and purified by metal-chelate affinity chromatography. The recombinant proteins were used as antigens in indirect enzyme-linked immunosorbent assay (ELISA) to differentiate between vaccinated and FMD-infected animals. The following parameters were determined: working concentrations of antigens and peroxidase conjugate of cattle anti-IgG, the optimum composition of blocking buffer, and the positive-negative threshold of the reaction. Tests performed with approximately 200 serum samples taken from animals of different immunity states showed that the protocol with protein 3A as the antigen (3A-ELISA) provided the most reliable differentiation. All the newly developed systems proved to outperform the commercial Chekit FMD-3ABC kit in sensitivity, and 3A-ELISA was no less specific.

[Immunological and biological characteristics of viral isolates of porcine reproductive and respiratory syndrome]. / Immunobiologicheskiie svoistva izoliatov virusa reproduktivnogo i respiratornogo sindroma svinei.

Described in the paper are study results of some immunobiological properties of 5 field isolates of the porcine reproductive and respiratory syndrome virus (PRRSV), obtained in Russia and Byelorussia in 1998-2001. According to the research work, the PRRSV isolates with different immunobiological properties have been circulating in the territories of Russia and Byelorussia. Three of the 4 examined virus isolates were found to be pathogenic and 1 avirulent to pigs.

A Clinical study of Intrauterine Fetal Death / 대한산부인과학회잡지

OBJECTIVE: The purpose of this study was to evaluate intrauterine fetal death and elucidate the etiology of intrauterine fetal death. METHODS: This is a clinical study of 74 cases of intrauterine fetal death (IUFD) among 5,523 deliveries at Soonchunhyang University Hospital during Jan. 1998 to Apr. 2003. RESULTS: The overall incidence of IUFD was 1.34%. And the age distribution of mother with IUFD was between 19 to 44 year old and was highest in the 25 to 29 year old age group (39.1%). The parity of mothers with IUFD was the highest in nulliparous group (78.3%) and there was a decreased tendency with high parity. There were 47 cases (63.5%) with previous history of abortion and 2 cases (2.7%) with previous history of IUFD. The highest incidence rate of IUFD was shown at 20-24 weeks of gestation (48.6%) and in the fetus weighted less than 1,000 gm (59.5%), and the sex ratio of male versus female fetus was 1:1.07. The modes of delivery were labor induction (54.1%), laparotomy (18.9%), spontaneous labor (27.0%). The indication for laparotomy was placental abruption, placenta previa, previous cesarean section state. The etiology factors of IUFD were unexplained causes (55.4%), cord complication (12.2%), placental abruption (9.4%), placenta previa (9.4%) in order. CONCLUSION: The causes of IUFD were unexplained, cord complication, placental abruption in order. So, the proper antenatal care should be taken of fetuses on the basis of risk factors of antepartum and intrapartum.

Renal effects of nabumetone, a COX-2 antagonist: impairment of function in isolated perfused rat kidneys contrasts with preserved renal function in vivo.

The constitutive cyclooxygenase (COX)-1 enzyme has been considered the physiologically important isoform for prostaglandin synthesis in the normal kidney. It has, therefore, been suggested that selective inhibitors of the 'inducible' isoform (COX-2) may be free from renal adverse effects. We studied the renal effects of the predominantly COX-2 antagonist nabumetone in isolated perfused kidneys. As compared with controls, kidneys removed after in vivo administration of oral nabumetone (15 mg/kg) disclosed altered renal function with reduced glomerular filtration rate, filtration fraction, and urine volume and enhanced hypoxic outer medullary tubular damage. By contrast, renal function and morphology were not affected in vivo by nabumetone or its active metabolite 6-methoxy-2-naphthylacetic acid. The latter agent (10-20 mg/kg i.v.) did not significantly alter renal microcirculation, as opposed to a selective substantial reduction in medullary blood flow noted with the nonselective COX inhibitor indomethacin (5 mg/kg i.v.). In a rat model of acute renal failure, induced by concomitant administration of radiocontrast, nitric oxide synthase, and COX inhibitors, the decline in kidney function and the extent of hypoxic medullary damage with oral nabumetone (80 mg/kg) were comparable to a control group, and significantly less than those induced by indomethacin. In rats subjected to daily oral nabumetone for 3 consecutive weeks, renal function and morphology were preserved as well. Both nabumetone and 6-methoxy-2-naphthylacetic acid reduced renal parenchymal prostaglandin E2 to the same extent as indomethacin. It is concluded that while nabumetone adversely affects renal function and may intensify hypoxic medullary damage ex vivo, rat kidneys are not affected by this agent in vivo, both in acute and chronic studies. COX selectivity may not explain the renal safety of nabumetone.

Compensated heart failure predisposes to outer medullary tubular injury: studies in rats.

BACKGROUND: Heart failure (HF) is considered a putative factor predisposing to acute renal failure (ARF). Since outer medullary hypoxic injury may play an important role in the pathogenesis of acute tubular necrosis, we explored the impact of experimental HF on the propensity to develop ARF with hypoxic medullary injury following the inhibition of prostaglandin and nitric oxide synthesis. METHODS: Compensated, high-output HF was induced in Sprague-Dawley rats by aorto-caval fistula. At the eighth to ninth postoperative day, the rats were injected with indomethacin and N(omega) nitro-L-arginine methyl ester (L-NAME; ARF protocol) and were sacrificed 24 hours later for morphologic evaluation. RESULTS: Kidney function comparably declined in HF-ARF rats and in control sham operated animals (CTR-ARF). Nevertheless, outer medullary hypoxic damage with medullary thick ascending limb (mTAL) necrosis occurred almost exclusively in the HF-ARF group (11 +/- 4% vs. 0.2 +/- 0.2% of tubules in CTR-ARF, P < 0.03). In a third group of HF animals subjected to vehicles only (HF-Nil), kidney function was preserved and renal morphology remained intact. Papillary-tip necrosis was consistently found in all animals subjected to indomethacin and L-NAME, irrespective of preconditioning. Morphometric evaluation disclosed that HF was not associated with mTAL hypertrophy. CONCLUSIONS: Incipient HF predisposes to hypoxic outer medullary injury, probably reflecting the impact of regional vasoconstrictive stimuli rather than tubular hypertrophy when protective local vasodilating mechanisms are hampered. The presence and extent of outer medullary hypoxic damage cannot be predicted from the functional derangement, which in the experimental settings may also represent prerenal azotemia or papillary damage.

Use of temporal temperature gradient gel electrophoresis to identify flaA and fim3 sequence types in Bordetella bronchiseptica.

AIMS: The aim of this study was to develop an approach to detect variation in the flaA and fim3 genes amongst animal isolates of Bordetella bronchiseptica using temporal temperature gradient gel electrophoresis (TTGE). METHODS AND RESULTS: Amplicons representing three flaA and two fim3 sequence types were subjected to TTGE analysis. It was possible to distinguish clearly between each of the sequence types using TTGE. CONCLUSION: This suggests that TTGE could be a useful tool for studying the epidemiology of B. bronchiseptica. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR amplification coupled to TTGE offers a general method for the rapid screening of large numbers of microbial strains for variations in gene sequences.

Transient urethral obstruction predisposes to ascending pyelonephritis and tubulo-interstitial disease: studies in rats.

Chronic tubulo-interstitial disease, an important cause of end-stage renal disease, often results from the combined effects of a disturbed urinary outflow tract and urinary tract infection. Acute unilateral ureteral obstruction in rats rapidly induces foci of medullary necrosis, confined to the region of the papilla and fornices. This injury may provide a nidus for bacterial invasion and may invoke reactive and regenerative changes, ultimately leading to chronic pyelonephritis and tubulo-interstitial nephropathy. To explore this possibility, adult rats underwent renal morphological evaluation 2-7 days following transient 24-h unilateral ureteral obstruction. In some experiments the bladder was inoculated with bacteria (10(8)-10(9) cfu/ml Escherichia coli in 0.5 ml) after release of ureteral obstruction, with subsequent cultures obtained from the pelvis of both kidneys and from the urinary bladder. Morphologic evaluation of perfusion-fixed kidneys, 2-7 days after the release of 24-h ureteral obstruction disclosed papillary necrosis, urothelial proliferation, marked inner-stripe interstitial expansion, and fibrosis and proximal tubular (S3) dilatation. The lateral (perihilar region) was predominantly affected, with lesions spreading from the fornices. There was some progression of interstitial fibrosis during the postobstructive time course or following more prolonged ureteral obstruction. By contrast, infection hardly contributed to the tubulointerstitial changes. In rats subjected to infection, cultures were positive in all 15 postobstructive kidneys, as opposed to five contralateral kidneys (P < 0.0001). Viable counts from the postobstructive kidney were also higher than those from the contralateral side (79,000 +/- 12,000 vs 2900 +/- 1600 cfu/ml, mean +/- SEM, P < 0.0001), and were comparable to those obtained from the bladder (77,000 +/- 13,000 cfu/ml). We conclude that transient ureteral obstruction predisposes to ascending pyelonephritis and to tubulointerstitial disease. This vulnerability may relate to altered urodynamics and medullary tissue destruction.