To investigate the mechanism by which prolactin (PRL) regulates follicular steroidogenesis in the ovary, we examined the functional roles of PRL in steroidogenesis using rat oocyte/granulosa cell coculture and focusing on the bone morphogenetic protein (BMP) system. The expression of long and short forms of PRL receptor (PRLR) were detected in both oocytes and granulosa cells, and PRL effectively up-regulated PRLR expression in granulosa cells in the presence of FSH. PRL suppressed FSH-induced estradiol production and increased FSH-induced progesterone production in granulosa cells. The PRL effects on FSH-induced progesterone were blocked by coculture with oocytes, implying roles of oocyte-derived factors in suppression of progesterone production in PRL-exposed granulosa cells. In accordance with the data for steroids, FSH-induced aromatase expression was suppressed by PRL, whereas FSH-induced steroidogenic acute regulatory protein, P450scc (P450 side-chain cleavage enzyme), and 3beta-hydroxysteroid dehydrogenase type 2 levels were amplified by PRL. However, forskolin- and N(6),O(2)-dibutyryl cAMP-induced steroid levels and FSH- and forskolin-induced cAMP were not affected by PRL, suggesting that PRL action on FSH-induced steroidogenesis was not due to cAMP-protein kinase A regulation. Treatment with a BMP-binding protein, noggin, facilitated PRL-induced estradiol reduction, and noggin increased PRL-induced progesterone production in FSH-treated granulosa cells cocultured with oocytes, suggesting that endogenous BMPs reduce progesterone but increase estradiol when exposed to high concentrations of PRL. PRL increased the expression of BMP ligands in oocyte/granulosa cell coculture and augmented BMP-induced phosphorylated mothers against decapentaplegic 1/5/8 signaling by reducing inhibitory phosphorylated mothers against decapentaplegic 6 expression through the Janus kinase/signal transducer and activator of transcription (STAT) pathway. In addition to STAT activation, PRL enhanced FSH-induced MAPK phosphorylation in granulosa cells, in which ERK activation was preferentially involved in suppression of FSH-induced estradiol. Furthermore, noggin treatment enhanced PRLR signaling including MAPK and STAT. Considering that BMPs suppressed PRLR in granulosa cells, it is likely that the BMP system in growing follicles plays a key role in antagonizing PRLR signaling actions in the ovary exposed to high concentrations of PRL.

ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs1) is an early immediate gene. We have previously reported ADAMTS1 was strongly induced by hypoxia. In this study, we investigated whether ADAMTS1 promoter-driven reporter signal is detectable by acute hypoxia. We constructed the GFP-expression vector (AHR-GFP) under the control of ADAMTS1 promoter and compared it with the constitutive GFP-expressing vector under the control of cytomegalovirus promoter (CMV-GFP). We transduced AHR-GFP and examined whether GFP signals can be detected under the acute hypoxia. When the human umbilical vein (HUVEC) was transduced under normoxia, there were few GFP signals, while CMV-GFP showed considerable GFP signals. When HUVEC was stimulated with hypoxia, GFP signals from AHR-GFP gene were induced under hypoxic conditions. Notably, the GFP signals peaked at 3h under hypoxia. In ischemic hind limb model, transduced AHR-GFP showed hypoxic induction of GFP signals. In summary, we demonstrated that AHR-system induced reporter gene expression by acute hypoxia, and its induction is transient. This is the first report showing the unique acute hypoxia-activated gene expression system.

Background: High-normal urinary albumin excretion has been reported to have clinical significance with respect to progression of proteinuria and hypertension. Objective: We analysed the effect of cilnidipine (10 mg/day) on morning systolic blood pressure (SBP) and urine albumin-creatinine ratio (UACR) in 16 non-diabetic hypertensive patients with a normal to marginally elevated UACR (mean +/- SD 29.4 +/- 21.7; range 7.5-72.9 mg/g creatinine). Methods: Sequential home BP and UACR data were fitted to a simple exponential function as follows: where y is SBP (mmHg) or UACR (mg/g creatinine); alpha is the extent of the SBP (mmHg)- or UACR (mg/g creatinine)-lowering effect; beta (days) is the time-constant for SBP or UACR decrease; t is the number of days after the start of cilnidipine administration; and gamma is the finally stabilized SBP (mmHg) or UACR (mg/g creatinine). Results: Mean +/- SD morning SBP and UACR decreased by 20.4 +/- 11.4 mmHg and 15.2 +/- 13.1 mg/g creatinine, respectively, as determined by coefficient alpha. The mean +/- SD time-constant for UACR decrease was significantly longer than that for BP decrease (43.5 +/- 22.9 vs 15.4 +/- 7.1 days). UACR reduction correlated with pre-treatment UACR values (correlation coefficient [R] = 0.88, p < 0.01) but not with BP decrease. Conclusions: The present study demonstrated that cilnidipine reduced UACR in hypertensive patients with normal to marginally elevated UACR independent of its BP-lowering effect.

OBJECTIVES: Based on well-established physiological theories, we studied correlations between changes in brachial-ankle pulse wave velocity (baPWV) relative to blood pressure (BP) elevation (elasticity of large-to-medium-sized arteries), and coronary artery disease (CAD). METHODS: The baPWV (in centimeters/second) and BP (in millimeters of mercury) were determined in 101 patients before, during, and/or after a cold pressor test using a volume-plethysmographic system. RESULTS: Significantly higher rates of increase in PWV relative to changes in BP were observed in the CAD(+) group than in the CAD(-) group when mean BP [median (25th-75th percentiles): 14.8 (8.3-24.9) vs. 8.6 (5.7-11.4) cm/s/mmHg, P<0.0001], and systolic [10.1 (6.0-17.5) vs. 6.4 (4.4-10.6) cm/s/mmHg, P=0.0023] and diastolic BP [21.0 (14.0-34.4) vs. 10.8 (6.8-16.1) cm/s/mmHg, P<0.0001] were used as BP indices. Similarly, the rates of increase in baPWV showed a significant correlation with the extent of CAD. The rate of increase in baPWV obtained using the mean, systolic and diastolic BP as indices showed an area under the receiver operating characteristic curve of 0.68-0.76, sensitivity of 65-75%, and specificity of 65-75% for the detection of CAD. The area under the receiver operating characteristic curve, sensitivity, and specificity for the rate of increase were slightly higher than those for baseline baPWV and baseline baPWV/baseline BP ratio, but not to a significant degree. CONCLUSION: The rate of increase in baPWV relative to BP elevation determined by cold pressor test is significantly and moderately correlated with CAD. To identify patients with CAD, the rate of increase in baPWV relative to changes in BP can provide considerable, but limited, information.

PURPOSE: To evaluate and compare the effect of short and long injection durations on aortic, pancreatic and hepatic enhancement in abdominal MDCT. METHODS AND MATERIALS: Triphasic contrast-enhanced CT images (16-MDCT, 1.25-mm collimation, 5-mm thickness, 6.1-s acquisition time for each phase) were obtained with 2mL/kg injection of 300mgI/mL iodine contrast material in 116 patients. Patients were prospectively randomized into two groups: one receiving contrast medium for 25-s injection duration and the other for 35-s injection duration. In both groups, triphasic scans were initiated 5, 15, and 40s after the completion of contrast injection for the first, second and third phases, respectively. CT values (HU) in the abdominal aorta, liver, spleen, pancreas, splenic and superior mesenteric arteries, and veins (splenic, superior mesenteric, portal, and hepatic) were measured. Quantitative and qualitative analysis for the degree of contrast enhancement between the two groups in various organs was compared at each scan phase. RESULTS: The aortic and arterial enhancements in the first-phase scan were higher for the 25-s group than those of the 35-s group (P<.001). Hepatic enhancement was higher for the 35-s group in the first (P<.001) and second (P<.01) phases, but no difference in the third-phase. No difference was found between the groups for the pancreatic enhancement at any phases. Qualitative results were in good agreement with quantitative results. CONCLUSION: Contrast administration with shorter injection duration increased peak aortic and arterial enhancement and contributed to improvement in the quality of CT angiograms, but for the solid abdominal organs 35-s protocol is recommended.

Heat-shock proteins (Hsps) are molecular chaperones that control protein folding and function. Argonaute 2 (Ago2), the effector in RNA interference (RNAi), is associated with Hsp90; however, its function in RNAi remains elusive. Here we show that Hsp90 is required for Ago2 to receive the small interfering RNA (siRNA) duplex from the RNA-induced silencing complex-loading complex in RNAi, suggesting a model where Hsp90 modifies Ago2 conformation to accommodate the siRNA duplex.

Imbalanced functions between osteoclasts and osteoblasts are involved in inflammatory bone damage. The clinical effectiveness of blocking TNF-alpha in treatment of active rheumatoid arthritis established the significance of TNF-alpha in the pathogenesis. In the present study, we investigated the cellular mechanism by which estrogen and glucocorticoid interact in osteoblastic differentiation regulated by BMP and TNF-alpha using mouse myoblastic C2C12 cells. The expression of estrogen receptors, (ER)alpha and ERbeta, and glucocorticoid receptor (GCR) was significantly increased by BMP-2 treatment regardless of the presence of estradiol and dexamethasone. Estradiol, but not dexamethasone, enhanced BMP-induced Runx2 and osteocalcin expression in C2C12 cells. In addition, TNF-alpha suppressed BMP-2-induced Runx2 and osteocalcin expression, and estradiol and dexamethasone reversed the TNF-alpha effects on BMP-2-induced Runx2 expression. Dexamethasone also abolished osteocalcin expression induced by BMP-2. Interestingly, BMP-2-induced Smad1/5/8 phosphorylation and Id-1 promoter activity were enhanced by estradiol pretreatment. On the other hand, dexamethasone suppressed BMP-2-induced Smad1/5/8 activation. TNF-alpha-induced SAPK/JNK activity was suppressed by estradiol, while NFkappaB phosphorylation was inhibited by dexamethasone. Of note, the inhibitory effects of TNF- on BMP-2-induced Runx2 and osteocalcin expression were reversed by SAPK/JNK inhibition regardless of the presence of estradiol. The estradiol effects that enhance BMP-2-induced Runx2 and osteocalcin mRNA expression were restored by antagonizing ER, and moreover, membrane-impermeable estradiol-BSA failed to enhance the BMP-2-induced osteoblastic differentiation. Thus, estrogen and glucocorticoid are functionally involved in the process of osteoblast differentiation regulated by BMPs and TNF-alpha. BMP-2 increases the sensitivities of ERs and GCR, whereas estrogen and glucocorticoid differentially regulate BMP-Smad and TNF-alpha signaling.

A 76-year-old man developed minimal-change nephrotic syndrome (NS). After treatment with prednisolone failed to induce sustained remission, cyclosporin was added. The NS improved, and prednisolone and cyclosporin doses were gradually decreased. However, he had repeated relapses of the syndrome, and at each relapse, the drug doses were increased. After 5 years, the patient developed left inguinal lymphadenopathy. The histological diagnosis was mixed cellularity classical Hodgkin lymphoma. He received 6 courses of ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine), and mixed cellularity classical Hodgkin lymphoma and NS both showed complete response. Although the association between Hodgkin lymphoma and minimal-change NS is well known, the pathogenesis is unknown. To the best of our knowledge, this is the first case report of minimal-change NS associated with Hodgkin lymphoma in which Hodgkin-Reed-Sternberg cells were immunostained for tumor necrosis factor-alpha (TNF-alpha) clearly demonstrating that Hodgkin-Reed-Sternberg produced TNF-alpha and in which the plasma level of TNF-alpha normalized after improvement of Hodgkin lymphoma by chemotherapy. The production of TNF-alpha by Hodgkin-Reed-Sternberg cells might play a key role as a potential mediator of minimal-change NS.

PURPOSE: To compare the results of emulsification of test materials when applying torsional and longitudinal emulsification tips as well as to define emulsification and determine how it occurs by examining the effect of phacoemulsification. SETTING: Miyoshi Eye Hospital, Fukuyama, Japan. METHOD: An ultra-high-speed digital video camera was used to record emulsification of chestnuts and human nuclei in a test chamber, simulating real phacoemulsification. The treatment of the nucleus as a result of phacoemulsification was analyzed visually. RESULTS: Visual comparison of video data showed that the test chamber materials were completely desiccated by the torsional phaco device. Longitudinal phacoemulsification resulted in cutting and coring of the test chamber materials, but not total disruption. CONCLUSIONS: Visual comparisons showed that complete obliteration and desiccation of the nucleus, or emulsification, occurs only with torsional phacoemulsification. These observations open the way to understanding the best approach to new tip design and surgical strategy development. FINANCIAL DISCLOSURE: Neither author has a financial or proprietary interest in any material or method mentioned.

MicroRNAs (miRNAs) are an abundant class of small non-coding RNAs that collectively regulate the expression of a large number of mRNAs by either promoting destabilization or repressing translation, or both. Therefore, they play a major role in shaping the transcriptomes and proteomes of eukaryotic organisms. Typically, animal miRNAs are produced from long primary transcripts with one or more of hairpin structures by two sequential processing reactions: one by Drosha in the nucleus and the other by Dicer in the cytoplasm. However, deviations from this paradigm have been observed: subclasses of miRNAs, which only partially meet the classical definition of a miRNA, are derived by alternative biogenesis pathways, thereby providing an additional level of complexity to miRNA-dependent regulation of gene expression.

OBJECTIVE: To determine how short-latency stretch reflex amplitude in the soleus muscle is modulated by cold stimulation in able-bodied individuals and individuals with complete spinal cord injury. METHODS: An initial 100-s baseline period was followed by 50-s cold stimulation periods. Stretch reflex of the right soleus muscle was elicited for 10-s intervals, while cold stimulation was applied to the left thigh. RESULTS: Peak-to-peak amplitude of the stretch reflex increased significantly during cold stimulation up to 127+/-21% of the baseline in the able-bodied group (n=9, P<0.01). Similarly, stretch reflex increased up to 125+/-11% in a group with injury level at or below thoracic 10 (n=4), although this increase was not significant. On the other hand, stretch reflex decreased significantly down to 78+/-20% in a group with injury level at or above thoracic 6 (n=8, P<0.05). CONCLUSIONS: Effect of afferent inputs induced by cold stimulation on stretch reflex modulation is different depending on the extent of central nervous systems participating in the modulation. SIGNIFICANCE: Our findings provide a better understanding of some basic changes in afferent-efferent spinal reflex pathways which are probably not monosynaptic in nature.

Heat shock protein 25 (Hsp25), which has anti-inflammatory activity, was examined for the relationship of its expression to macrophage appearance in thioacetoamide (TAA)-induced rat acute hepatic lesions. TAA-induced lesions, consisting of hepatocyte coagulation necrosis and reactive macrophages, developed in the centrilobular areas. Macrophages immuno-reacting to ED1 (CD68; exudative macrophages) were mainly seen within the lesions, whereas macrophages reacting to ED2 (CD163; resident macrophages and Kupffer cells), which have abundant cytoplasm, appeared mainly in the periphery of the lesions. Hsp25-immunopositivity was seen in hepatocytes around the lesions in relation to ED1- and ED2-positive macrophages in and around the centrilobular lesions, respectively. Because macrophages appearing in early stages of hepatic lesions produce various pro-inflammatory factors, mRNA expressions of tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant factor-1 (MCP-1) and osteopontin (OPN) were examined in relation to Hsp25 mRNA expression. Hsp25 mRNA expression generally was correlated with TNF-alpha, MCP-1 and OPN expressions, suggesting their direct or indirect association with Hsp25 expression. Thus, Hsp25 might have a cytoprotection function against macrophages appearing in hepatic lesions, and factors produced by macrophages in the very early stages of hepatic lesions may influence Hsp25 expression. Hsp25 expression should be useful as an index of anti-inflammatory action for evaluation of hepatotoxicants in vivo.

Summary Sepsis is a generalized inflammatory disease, caused by the hyperinflammatory response of the host, rather than by invading organisms. Endothelial cells play a crucial role in the pathogenesis of sepsis. In this study, we investigated the effects of interleukin-8 (IL-8), a known neutrophil chemoattractant, on lipopolysaccharide (LPS) -induced reactive oxygen species (ROS) production by endothelial cells, and its significance in the pathogenesis of LPS-mediated sepsis. The results revealed that IL-8 directly induced ROS production in human umbilical vein endothelial cells (HUVECs), and also mediated LPS-induced ROS production by HUVECs. Stimulation of HUVECs by LPS strongly enhanced tissue factor expression, a hallmark of severe sepsis, which was suppressed by IL-8 knockdown. We further discovered that NADPH oxidase (Nox) 1 expression in LPS-stimulated HUVECs was markedly repressed by IL-8 knockdown, and Nox1 knockdown reduced tissue factor expression, suggesting that the LPS/IL-8 signalling in endothelial cells was predominantly mediated by Nox1. In conclusion, LPS stimulation of endothelial cells causes activation of the IL-8-Nox1 axis, enhances the production of ROS, and ultimately contributes to the progression of severe sepsis.

Case 1: An 86-year-old woman had an invasive breast cancer with dermal infiltration. Bone metastases were found in the femur and lumbar vertebrae. She was treated with 2 courses of 70 mg docetaxel(DOC)chemotherapy every 3 weeks, after which the tumor dramatically decreased in size. Following this treatment, she underwent a radical mastectomy. Case 2: An 80-year-old woman had a 10 cm tumor in the right breast. Lung and bone metastases were also found. Two 80 mg courses of DOC reduced the lung and bone metastases, and the size of the breast tumor. She underwent a local excision. Hormonal therapy is a standard treatment for hormone-sensitive breast cancer in elderly patients. It is suitable for patients who have a declining quality of life(QOL), although chemotherapy shows a higher response rate and takes less time than hormonal therapy. However, it is difficult to continue chemotherapy until pCR is achieved, even for chemotherapy-effective patients because the side effects of chemotherapy are severe. Therefore, local excision after chemotherapy is necessary for breast cancer patients to improve their QOL, even if there are distant metastatic lesions.

Alkynyl oxime ether underwent a gold-catalyzed domino reaction involving cyclization and subsequent Claisen-type rearrangement to afford trisubstituted isoxazoles in a direct, efficient, and regioselective manner. The products were successfully applied to the synthesis of unusual heterocycles as an illustration of the potential utility of the reaction.

Bone morphogenetic proteins (BMPs) have been recognized as crucial molecules in regulating ovarian physiology, with different BMPs having differential actions in FSH-induced estradiol production. To identify the roles of oocyte factors that modulate steroidogenesis controlled by BMPs, we here investigated the effects of FGF-8 in rat granulosa/oocyte co-cultures. FGF-8 potently suppressed FSH-induced estradiol production, but did not affect cAMP-induced estradiol produced by rat granulosa cells. FGF-8 had no effects on progesterone and cAMP production induced by FSH and forskolin. The inhibitory effects of FGF-8 on FSH-induced estradiol production were not altered by BMP-2, -4, -6 or -7. In the presence of FGF-8, BMPs suppressed FSH-induced progesterone by reducing cAMP, suggesting that FGF-8 and BMP independently regulate FSH receptor signaling. Notably, FGF-8 induced ERK and SAPK/JNK phosphorylation in granulosa cells, in which ERK activation was further enhanced by FSH and oocytes. Inhibition of ERK and SAPK/JNK reduced FSH-induced progesterone and cAMP levels, suggesting that the activation of these pathways enhances FSH-induced cAMP signaling. In addition, ERK inhibition upregulated FSH-induced estradiol synthesis, indicating that ERK pathway is also involved in suppressing aromatase activity in granulosa cells. Interestingly, FGF-8 enhanced BMP-induced Smad1/5/8 and Id-1-promoter activities with decreased expression of Smad6/7. Since the SAPK/JNK inhibitor inhibited FGF-8 effects in upregulating Id-1 transcription, SAPK/JNK appears to be involved in the mechanism by which FGF-8 enhances BMP-Smad signaling. Furthermore, in the presence of oocytes, the inhibition of endogenous FGF receptor signaling suppressed FSH- and forskolin-induced progesterone and cAMP, showing that endogenous FGF system is involved in activation of FSH-induced cAMP-PKA signaling via ERK and SAPK/JNK. Thus, the oocyte factor, FGF-8, not only suppresses FSH-induced estradiol production by activating ERK, but also enhances BMP-Smad signaling in granulosa cells. This interaction between FGF-8 and BMPs may play a key role in regulating steroidogenesis through oocyte-granulosa cell communication.

This study aimed to examine the impact of membrane properties on membrane fouling in membrane bioreactor (MBR). Membrane fouling was divided into two categories: physically reversible and irreversible fouling. Membrane properties related to each type of membrane fouling were investigated separately. Five microfiltration (MF) and one ultrafiltration (UF) membranes with different properties (pore size, contact angle, roughness, zeta potential, and pure water permeability) were examined with a laboratory-scale MBR, fed with synthetic wastewater. Two separate experiments were conducted: the first to examine physically reversible fouling, and the second to examine physically irreversible fouling. The correlation between the degree of each type of fouling and membrane properties was studied. High correlation was observed between the degree of physically reversible fouling and roughness (R(2)=0.96). In contrast, with regard to physically irreversible fouling, strong correlation between roughness and degree of membrane fouling can only be found in the case of MF membranes. Except for the membrane with the highest roughness, the degree of physically irreversible fouling can be well correlated with pure water permeability (lower pure water permeability results in higher degree of physically irreversible fouling) including UF membrane. On the basis of the results obtained in this study, it can be concluded that roughness is an important factor in determination of physically reversible fouling regardless of the types of membrane (i.e. MF or UF membranes) and evolutions of physically irreversible fouling can be mitigated when an MBR is operated with membranes with smooth surface and high pure water permeability.

In this study, the fouling potentials of polysaccharides contained in mixed liquor suspension in a membrane bioreactor (MBR) treating municipal wastewater were investigated using lectin affinity chromatography. This investigation was carried out with different membranes to assess the effects of membrane materials on fouling potentials of polysaccharides. In lectin affinity chromatography, some polysaccharides with high affinity to the lectin in the column can be retained in the column. The fouling potentials of the retained polysaccharides were evaluated by dead-end filtration test. Degree of reduction in fouling potential differed considerably when different lectins were used in affinity chromatography indicating that fouling potentials of polysaccharide differed depending on types of polysaccharide. Trends in the reduction of fouling potential were different depending on membrane materials. Characteristics of the organic matter associated with polysaccharides removed by lectins were investigated by means of excitation-emission matrices (EEM). The results of EEM analysis indicate that the characteristics of the organic matter eluted from different lectins were different as long as elution reagents for the lectin were different. Characteristics of the organic matter eluted from the lectins which have the same elution reagent were similar in terms of shapes of EEM fluorescence spectra. However, the trends in reduction of fouling potentials could not be explained by the characteristics of organic matter assessed by EEM analysis. On the basis of the results obtained in this study, it can be concluded that characteristics of membrane and structures or properties of sugar chain would play an important role in determining fouling potentials.