Metabolic biomarkers for non-alcoholic fatty liver disease induced by high-fat diet: In vivo magnetic resonance spectroscopy of hyperpolarized [1-13C] pyruvate.

Hyperpolarized 13C magnetic resonance spectroscopy (MRS) to assess hepatic metabolism in non-alcoholic fatty liver disease (NAFLD) has not been reported. This study searched for cellular metabolism-based biomarkers for NAFLD induced by a high-fat diet (HFD) in rats. Also, correlations of the biomarkers with enzyme levels and histopathology were identified during a 6-week follow-up. Six rats were fed a control diet (CD) and seven rats were fed the HFD for 6 weeks. Hyperpolarized 13C dynamic MRS was performed on rat liver following an injection of hyperpolarized [1-13C] pyruvate. Compared with CD-fed rats, HFD-fed rats showed significant increases in the levels of serum alanine aminotransferase and low-density lipoprotein cholesterol at weeks 4 and 6 of follow-up. After the 6-week HFD, the ratios of [1-13C] alanine/pyruvate and [1-13C] lactate/pyruvate were significantly increased, as were the levels of alanine aminotransferase and lactate dehydrogenase, which are potentially associated with hepatosteatosis. The results implicate [1-13C] alanine and [1-13C] lactate as potentially useful noninvasive biomarkers of hepatosteatosis occurring in NAFLD.

All-Trans Retinoic Acid Increases Aquaporin 3 Expression in Human Vaginal Epithelial Cells.

INTRODUCTION: Water channel aquaporin 3 (AQP3) is an aquaglyceroporin that transports small neutral solutes and water. All-trans retinoic acid (ATRA), a member of the retinoid drug class, acts as a regulator in several biological processes. AIM: To investigate the effect of ATRA on the expression of AQP3 in human vaginal epithelial cells. METHODS: Human vaginal mucosal epithelial cells (CRL2616) were treated with ATRA 0, 0.01, 0.1, and 1 µmol/L for 24 hours to examine the dose-dependent effects of ATRA and with ATRA 1 µmol/L for 0, 3, 6, 12, and 24 hours. MAIN OUTCOME MEASURES: The expression of AQP3 and retinoic acid receptor (RAR) was determined by western blot analysis and reverse transcription polymerase chain reaction. RESULTS: AQP3 was detected in the cell membrane of human vaginal epithelial cells. ATRA increased the protein expression and mRNA levels of AQP3 in a dose-dependent manner (P < .05). ATRA also increased the protein expression of RARα (P < .05). Treatment of CRL2616 cells with an RAR antagonist (Ro 41-5253) significantly decreased AQP3 protein expression (P < .05). CONCLUSION: ATRA mediated by RARα increased AQP3 gene and protein expression in human vaginal mucosal epithelial cells. These results imply that AQP3 regulated by ATRA could play an important role in the mechanism of vaginal lubrication.

Time-course metabolic changes in high-fat diet-induced obesity rats: A pilot study using hyperpolarized (13)C dynamic MRS.

The purpose of this study was to investigate the time-course metabolic changes based on hyperpolarized (13)C magnetic resonance spectroscopy (MRS) in high-fat diet (HFD)-induced obesity rats and the correlation between metabolic and serum enzyme levels. Sprague-Dawley rats were fed either HFD (60% fat) or normal diet (10% fat) for 6weeks. A HyperSense DNP was used to hyperpolarize [1-(13)C] pyruvic acid and the hyperpolarized (13)C MRS was examined every 2weeks in the course of 6weeks using a 3T GE MR750 scanner. The body weight of HFD-induced obese rats was significantly increased compared to normal rats at the 6th week after the onset of feeding (p=0.05). Simultaneously, the HFD-induced obese rats showed significantly increased levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and low-density lipoprotein (LDL)-cholesterol compared to normal rats (p≤0.05). In the dynamic (13)C MR spectra acquired at the 6th week, the obese rats showed significantly increased ratios of [1-(13)C] lactate/[1-(13)C] pyruvate and [1-(13)C] alanine/[1-(13)C] pyruvate (p=0.05). The (13)C spectral outcomes are positively correlated with the enzyme levels of ALT and LDH in the HFD-induced obesity. The [1-(13)C] lactate and [1-(13)C] alanine are potentially considered as noninvasive biomarkers for the HFD-induced obesity.

Estrogen Modulates Expression of Tight Junction Proteins in Rat Vagina.

Background. The objectives of this study were to investigate the localization of tight junctions and the modulation of zonula occludens- (ZO-) 1, occludin and claudin-1 expression by estrogen in castrated female rat vagina. Female Sprague-Dawley rats (230-240 g, n = 45) were divided into three groups and subjected to a sham operation (control group, n = 15), bilateral ovariectomy (Ovx group, n = 15), or bilateral ovariectomy followed by daily subcutaneous injection of 17ß-estradiol (50 µg/kg/day, Ovx + Est group, n = 15). The cellular localization and expression of ZO-1, occludin, and claudin-1 were determined in each group by immunohistochemistry and western blot. Results. Expression of ZO-1 was diffuse in all groups, with the highest intensity in the superficial epithelium in the control group. Occludin was localized in the intermediate and basal epithelium. Claudin-1 was most intense in the superficial layer of the vaginal epithelium in the control group. Expression of ZO-1, occludin, and claudin-1 was significantly decreased after ovariectomy and was restored to the level of the control after estrogen replacement. Conclusions. Tight junctions are distinctly localized in rat vagina, and estrogen modulates the expression of tight junctions. Further researches are needed to clarify the functional role of tight junctions in vaginal lubrication.

COMP-angiopoietin-1 promotes cavernous angiogenesis in a type 2 diabetic rat model.

Cartilage oligomeric matrix protein-angiopoietin-1 (COMP-Ang1) is an angiogenic factor for vascular angiogenesis. The aim was to investigate the effect of an intracavernosal injection of COMP-Ang1 on cavernosal angiogenesis in a diabetic rat model. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats made up the experimental group (1 yr old) and Long-Evans Tokushima Otsuka (LETO) rats made up the control group. The experimental group was divided into vehicle only, 10 µg COMP-Ang1, and 20 µg COMP-Ang1. COMP-Ang1 was injected into the corpus cavernosum of the penis. After 4 weeks, the penile tissues of the rats were obtained for immunohistochemistry and Western blot analysis. The immunoreactivity of PECAM-1 and VEGF was increased in the COMP-Ang1 group compared with the vehicle only group. Moreover, the expression of PECAM-1 and VEGF was notably augmented in the 20 µg Comp Ang-1 group. In the immunoblotting study, the expression of PECAM-1 and VEGF protein was significantly less in the OLEFT rats than in the control LETO rats. However, this expression was restored to control level after intracavernosal injection of COMP-Ang1. These results show that an intracavernosal injection of COMP-Ang1 enhances cavernous angiogenesis by structurally reinforcing the cavernosal endothelium.

Effect of hyperglycemia on expression of aquaporins in the rat vagina.

OBJECTIVE: To investigate the effect of hyperglycemia on the expression of the aquaporin (AQP) isoforms in the diabetic rat vagina. METHODS: Female Sprague-Dawley rats (230-240 g, n = 45) were divided into control (n = 10) and experimental (n = 35) groups. Diabetes in the experimental group was induced by intraperitoneal injection of streptozotocin (STZ, 65 mg/kg). STZ-induced diabetic rats were left untreated or given subcutaneous injections of insulin (3 U/d). After 2 and 4 weeks, the blood glucose was measured, and the vaginal blood flow was assessed by Doppler flowmetry. The expression and cellular localization of AQP1 and AQP2 in the rat vagina were determined by Western blot and immunohistochemistry. RESULTS: The vaginal blood flow (mL/min/100 g tissue) after pelvic nerve stimulation was significantly lower in the STZ-induced diabetic rats (21.9 ± 6.5 at 2 weeks and 21 ± 2.8 at 4 weeks) compared with the control group (55.5 ± 8.9 at 2 weeks and 52.9 ± 6.5 at 4 weeks; P < .05). In contrast, the vaginal blood flow response was significantly greater in the insulin-treated diabetic groups (47.7 ± 8.7 at 2 weeks and 47.7 ± 8.4 at 4 weeks) and comparable to that of the control group. The protein expression of AQP2 was significantly lower in the STZ-induced diabetic rats and was restored to the control level after insulin treatment. However, no change was seen in AQP1 expression. Thus, hyperglycemia might cause downregulation of AQP2 expression in the diabetic rat vagina. CONCLUSION: These results suggest that decreased vaginal lubrication in diabetic women might result from changes in aquaporin expression, in addition to a reduction in the vaginal blood flow response.

Changes in aquaporin (AQP)2 and AQP3 expression in ovariectomized rat urinary bladder: potential implication of water permeability in urinary bladder.

PURPOSE: AQPs have recently been reported to be expressed in rat and human urothelium. The purpose of this study was to investigate the effect of ovariectomy on the expression of AQP2 and AQP3 in rat urothelium. MATERIALS AND METHODS: Female Sprague-Dawley rats were divided into three groups: control, bilateral ovariectomy (Ovx), and bilateral ovariectomy followed by subcutaneous injections of 17ß-estradiol (Ovx + Est). After 4 weeks, urodynamic studies were done to measure the contraction interval and contraction pressure. The expression and cellular localization of AQP2 and AQP3 were determined by Western blot and immunohistochemistry in rat urinary bladder. RESULTS: In cystometrograms, the contraction interval (min) was significantly lower in the Ovx group (2.8 ± 0.32) than in the control group (5.1 ± 0.56) but was increased after estrogen treatment (8.8 ± 0.29). Conversely, the average contraction pressure (mmHg) was higher in the Ovx group (28.2 ± 2.3) than in the control group (22.3 ± 1.06) and decreased after estrogen treatment (23.1 ± 2.02). AQP2 expression was localized in the cytoplasm of the epithelium, whereas AQP3 was found only in the cell membrane of the epithelium. The protein expression of both AQP2 and AQP3 was significantly lower after ovariectomy and was restored to the control levels after 17ß-estradiol treatment. CONCLUSIONS: Hormonal alteration causes a significant change in the expression of AQP2 and AQP3. These findings suggest that AQPs might have a functional role in the detrusor overactivity that occurs in association with hormonal alteration in female rat.

Tear production and ocular surface changes in experimental dry eye after elimination of desiccating stress.

PURPOSE: To investigate the severity and duration of desiccating stress-induced dry eye disease between mice with and without a genetic predisposition to spontaneous autoimmunity. METHODS: Experimental dry eye was induced in 12- to 16-week-old wild-type C57BL/6 and autoimmune NOD.B10.H2(b) mice by subcutaneous injection of scopolamine with exposure to an air draft for 10 days. Tear volume and corneal smoothness were measured at baseline, 5 and 10 days after desiccating stress, and 3, 7, 14, and 28 days after the removal of desiccating stress. Periodic acid-Schiff staining and immunohistochemistry were performed to evaluate the densities of conjunctival goblet cells and CD4(+) T cells in each group. Interleukin (IL)-1ß and IL-6 concentrations in conjunctival tissues were measured by multiplex immunobead assay. RESULTS: Signs of experimental dry eye were noted at 5 and 10 days after desiccating stress in both strains. After the removal of desiccating stress, in C57BL/6 mice, tear production and corneal smoothness improved at 3 and 7 days, respectively, and conjunctival goblet cells and CD4(+) T-cell densities and cytokine levels returned to baseline levels at 14 days. In contrast, in NOD.B10.H2(b) mice, none of the parameters recovered to baseline levels during a period of 28 days after the removal of desiccating stress. CONCLUSIONS: After the removal of desiccating stress in experimental dry eye, tear volume and ocular surface parameters recovered within 2 weeks in C57BL/6 mice, whereas they remained unchanged in NOD mice. In contrast to autoimmune mice, experimental dry eye can be reversed after the elimination of desiccating stress in nonsusceptible mice.

Expression of aquaporin water channels in the vagina in premenopausal women.

INTRODUCTION: Aquaporins (AQPs) are membrane proteins that facilitate water movement across biological membranes. This study builds on a previous report on the distinct localization of AQPs in the rat vagina. AIM: The purposes of this study were to investigate the localization and expression of the AQPs in the vaginal tissue of premenopausal women. METHODS: Anterior vaginal tissue was collected during transvaginal uterine myomectomy or hysterectomy from 10 premenopausal women (mean age, 40 years) for Western blot and immunohistochemistry. MAIN OUTCOME MEASURES: The expression and cellular localization of AQP1-9 were determined in the human vagina by Western blot and immunohistochemistry. RESULTS: Immunolabeling showed that AQP1 was mainly expressed in the capillaries and venules of the vagina, AQP2 was expressed in the cytoplasm of the epithelium, AQP3 was mainly associated with the plasma membrane of the vaginal epithelium, and both AQP5 and AQP6 were expressed in the cytoplasm throughout all vaginal epithelium. Western blot analysis revealed bands at 28 kDa for AQP1, 2, 3, 5, and 6 proteins. However, AQP4, 7, 8, and 9 were not detected. CONCLUSIONS: The distinct localization of AQPs in the human vagina suggests that AQP1, 2, 3, 5, and 6 may play an important role in vaginal lubrication in women.

Distribution of interstitial cells of cajal in menopausal rat urinary bladder showing detrusor overactivity.

PURPOSE: Recent studies have showed that interstitial cells of Cajal (ICCs) are widely distributed in the genitourinary tract and have suggested their involvement in spontaneous electrical activity and muscle contraction. The purposes of this study were to investigate the effect of estrogen on ICCs in rat urinary bladder from the detrusor overactivity induced by ovariectomy. MATERIALS AND METHODS: Female Sprague-Dawley rats (230-240 g, N=60) were divided into three groups: control (N=20), bilateral ovariectomy (Ovx, N=20), and bilateral ovariectomy followed by subcutaneous injections of 17 ß-estradiol (50 mg/kg/day, Ovx + Est, N=20). After 4 weeks, urodynamic studies measuring contraction interval and contraction pressure were done. The cellular localization of ICCs was determined by immunohistochemistry in the rat urinary bladder. RESULTS: Filling cystometry studies demonstrated a reduced interval between voiding contractions and an increased voiding pressure in Ovx group. The approximate the contraction interval (min) was (3.9±0.25) significantly decreased in the Ovx group compared to the control group (6.7±0.15), which was increased after estrogen treatment (9.7±0.22) (p<0.05). Conversely, the average contraction pressures (mmHg) were increased in the Ovx group (28.9±2.1) compared to the control group (21.2±1.45), and decreased after estrogen treatment (24.8±2.21) (p<0.05). The population of c-Kit immunoreactive ICCs was decreased in both the urothelial and muscle layers in Ovx bladders, which increased to the control value after estrogen treatment. CONCLUSIONS: These results demonstrated an decreased immunoreactivity of ICCs in the menopausal rat model and suggest that thedecreased population of ICCs expression may contribute to the modulation of bladder overactivity induced by menopause.

The Expression of AQP1 and eNOS in Menopausal Rat Urinary Bladder.

PURPOSE: Aquaporins (AQPs) have been reported to be expressed in rat and human urothelium. Nitric oxide (NO) is thought to play an important role in the bladder overactivity related to menopause. The purpose of this study was to investigate the effect of hormonal alteration on the expression of AQP1 and eNOS in menopausal rat urinary bladder. MATERIALS AND METHODS: Female Sprague-Dawley rats (230-240 g, N=30) were divided into three groups: control (N=10), bilateral ovariectomy (Ovx, N=10), and bilateral ovariectomy followed by subcutaneous injections of 17ß-estradiol (50 mg/kg/day, Ovx+Est, N=10). After 4 weeks, urodynamic studies measuring the contraction interval and contraction pressure were done. The expression and cellular localization of AQP1 and eNOS were determined by performing Western blotting and immunohistochemistry on the rat urinary bladder. RESULTS: The approximate contraction interval (min) was significantly decreased in the Ovx group (3.9±0.25) compared to the control group (6.7±0.15), and was increased after estrogen treatment (9.7±0.22) (p<0.05). The AQP1 and eNOS immunoreactivities were localized in the same areas: capillaries, arterioles, and venules of the lamina propria. The protein expression of AQP1 was not changed significantly, whereas eNOS expression was significantly decreased in the Ovx group and restored to the control value in the Ovx+Est group. CONCLUSIONS: This study showed that ovariectomy causes a significant change in e-NOS expression without a change in AQP1 in menopausal rat urinary bladder. This may imply that e-NOS has a functional role in the bladder overactivity that occurs in association with menopause.

Effect of estrogen deprivation on the expression of aquaporins and nitric oxide synthases in rat vagina.

INTRODUCTION: The expression of aquaporin (AQP) water channels in rat vagina was recently reported. AIM: The purposes of this study were to investigate the effect of 17beta-estradiol on the expression of the AQP-1 and AQP-2 water channels and nitric oxide synthase (NOS) isoforms in rat vagina. METHODS: Female Sprague-Dawley rats (230-240 g, N = 90) were divided into three groups: control (N = 30), bilateral ovariectomy (N = 30), and bilateral ovariectomy, followed by subcutaneous injections of 17beta-estradiol (50 microg/kg/day, N = 30). After 4 weeks, genital hemodynamics and vaginal secretions were measured after pelvic nerve stimulation, and the animals were then killed. The expression and cellular localization of AQP-1, AQP-2, endothelial NOS (e-NOS), and neuronal NOS (n-NOS) were determined in each group by immunohistochemistry and Western blot. MAIN OUTCOME MEASURES: The expression and cellular localization of AQPs and NOS isoforms after estrogen deprivation. RESULTS: Estimated vaginal secretions (mg, mean +/- standard error) were significantly lower in the ovariectomized group (2.9 +/- 0.62) than in the control group (5.7 +/- 1.25) and returned to the control value in the group after treatment with 17beta-estradiol (6.5 +/- 1.22) (P < 0.05). Both AQP-1 and e-NOS immunoreactivities were localized in the capillaries and venules of the lamina propria of the vagina, and n-NOS was expressed in the nerve fibers of the subepithelial lamina propria. The expression of AQP-2 was localized solely in the superficial layer of the vaginal epithelium. The protein expressions of AQP-2, e-NOS, and n-NOS were significantly lower after ovariectomy and were restored to the control level after 17beta-estradiol treatment. However, there was no significant change in AQP-1 expression. CONCLUSIONS: Decreased vaginal secretion after estrogen deprivation may be partly due to functional changes in both AQPs and NOS isoforms in the vagina. The potential role of AQPs in water transport in the vagina might differ according to the type of AQP.

Expression of aquaporin water channels in rat vagina: potential role in vaginal lubrication.

INTRODUCTION: Aquaporins (AQPs) are membrane proteins that facilitate water movement across biological membranes. There has been little research on the role of AQPs in the female sexual arousal response. AIM: The purposes of this study were to investigate the localization and functional roles of AQP1, AQP2, and AQP3 in rat vagina. METHODS: Female Sprague-Dawley rats (230-240 g, N = 20) were anesthetized. The vaginal branch of the pelvic nerve was stimulated for 60 seconds (10 V, 16 Hz, 0.8 ms), and the animals were sacrificed either immediately or 5 minutes later. The expression and cellular localization of AQP1, 2, and 3 were determined by Western blot and immunohistochemistry of the vagina. The intracellular membrane and plasma membrane fractions of the proteins in vaginal tissue were studied by immunoblot analysis with the differential centrifugation. MAIN OUTCOME MEASURES: The expression and cellular localization of AQPs, and pelvic nerve stimulation induced translocation of AQPs in rat vaginal tissue. RESULTS: Immunolabeling showed that AQP1 was mainly expressed in the capillaries and venules of the vagina. AQP2 was expressed in the cytoplasm of the epithelium, and AQP3 was mainly associated with the plasma membrane of the vaginal epithelium. AQPs were found to be present primarily in the cytosolic fraction of untreated tissues. The translocation of AQP1 and 2 isoforms from the cytosolic compartment to the membrane compartment was observed immediately after nerve stimulation and had declined at 5 minutes after nerve stimulation, while the subcellular localization of AQP3 was not changed by nerve stimulation. CONCLUSIONS: These results showed a distinct localization of AQPs in the rat vagina. Pelvic nerve stimulation modulated short-term translocation of AQP1 and 2. These results imply that AQPs may play an important role in vaginal lubrication.

Polyion complex micelles composed of all-trans retinoic acid and poly (ethylene glycol)-grafted-chitosan.

The goal of this study is to develop novel types of polyion complex micelles for the drug delivery to brain tumor. Methoxy poly(ethylene glycol) (mPEG)-grafted chitosan (CP) was synthesized in order to make polymeric micelles encapsulating all-trans retinoic acid (ATRA) based on polyion complex formation. Polyion complex micelles were found to have spherical shapes with sizes of about 50 approximately 200 nm. The loading efficiency of micelle was higher than 80% (w/w) for all formulations. 1H nuclear magnetic resonance (NMR) spectra confirmed the formation of polymeric micelles. The CP graft copolymer and ATRA have distinguishing peaks in their 1H NMR spectra. The specific peaks of ATRA disappeared in D2O or DMSO while it appeared at mixtures of D2O/DMSO, indicating that ATRA and chitosan formed ion complex inner-core. In the cell cytotoxicity study using U87MG cells in vitro, polyion complex micelles showed similar cytotoxicity to that of free ATRA. A migration test was performed to investigate the inhibition of tumor cell invasion in vitro. The results suggested that the polyion complex micelles was more effective at inhibiting tumor cell migration than free ATRA.

Expression of Nm23 in gliomas and its effect on migration and invasion in vitro.

BACKGROUND: Despite the well-documented importance of Nm23 in the control of metastasis, there is currently a paucity of data regarding the role of this gene family in the control of glioma invasion. MATERIALS AND METHODS: Nm23-H1 expression in gliomas was assessed via immunohistochemistry, Western blot, RT-PCR and Northern blot analyses. The migration and invasion ability were also investigated in primary glioma culture cells, human glioma cell lines and nm23-H1 transfectant, using an in vitro brain slice invasion model and a simple scratch technique. RESULTS: Although no significant correlations were detected between nm23-H1 expression and pathological grade, the endogenous nm23-H1 expression in gliomas was found to be inversely correlated with their migratory abilities. Additionally, the nm23-H1 transfectant resulted in a reduction of approximately 45% of the migratory ability and suppressed the invasiveness of the parental cell line. CONCLUSION: Our overall findings suggest that nm23-H1 may play an important role in the suppression of glioma invasion and migration.

Preparation of poly(DL-lactide-co-glycolide) microspheres encapsulating all-trans retinoic acid.

Poly(DL-lactide-co-glycolide) (PLGA) microspheres containing all-trans retinoic acid (atRA) were prepared by o/w solvent evaporation method and various preparation parameters, such as poly(vinyl alcohol) (PVA) concentration in aqueous solution, PVA MW, drug weight, solvent, polymer MW, and polymer weight, on the characteristics of microspheres and drug release were investigated. PVA concentration in water phase was a critical factor in making microspheres consistently with smooth surface and round shape. In our study, at least 2% (w/v) of PVA in aqueous solution was necessary for making microspheres with round shape. The particle size of microspheres ranged 10-100 microm. AtRA was slowly released from PLGA microspheres over 30 days. Sterilization of microspheres by ethylene oxide (EO) gas at 37 degrees C did not significantly affect the characteristics of drug release or its morphology. Cell growth inhibition of atRA was affected by preparation process of microspheres rather than the EO-gas sterilization process. These results indicate that PLGA microspheres containing atRA are acceptable for controlled release devices for use in the treatment of brain tumor.

Brain tumor invasion model system using organotypic brain-slice culture as an alternative to in vivo model.

PURPOSE: The primary cause of local recurrence and therapeutic failure in the treatment of malignant gliomas is the invasion of tumor cells into the surrounding normal brain. While it is known that malignant gliomas infiltrate diffusely into regions of normal brain, it is frequently very difficult to unequivocally identify the solitary invading glioma cell in histopathological preparations, or in experimental glioma models. We have developed an experimental invasion assay system, which allows us to track the solitary invasive glioma cell, using human brain tissue obtained from routine craniotomies for seizures or trauma. METHODS: This tissue is cut into 1-mm thick slices and cultured in the upper chamber of Transwell culture dishes on top of a 0.4- micro m pore size polyester membrane, which is fed on medium provided in the lower chamber. Glioma cells are stably transfected with vectors containing a green fluorescent protein (GFP) cDNA. Stable, high-level expression GFP transfectants were selected by direct visualization under fluorescence microscope. In addition, various tumor spheroids are stained with vital dye, DiI, to track the invading cells. GFP-expressing glioma cells or stained spheroids were then implanted on the center of the brain slice, and the degree of brain tumor invasion into the brain tissue was evaluated at different time points by optical sectioning using a confocal microscope. RESULTS: We observed that GFP-expressing glioma cells or stained spheroids could be readily tracked and followed with this model system. Individual tumor cells that exhibited green or red fluorescence could be identified and their migration path through the brain slices unequivocally followed. CONCLUSION: This experimental invasion system may be of considerable utility in studying the process of brain tumor invasion and in evaluating its invasiveness in individual brain tumor because it not only provides a better representation of extracellular matrix molecules normally encountered by invading glioma cells, but also provides the fluorescent tag applied to the tumor cells.

Diabetes induced alteration of clitoral hemodynamics and structure in the rabbit.

PURPOSE: We investigated the effect of diabetes on clitoral hemodynamics and structures in the rabbit. MATERIALS AND METHODS: A total of 25 New Zealand White female rabbits weighing 3 to 3.5 kg. were divided into 2 groups, including 5 in the control and 20 in the experimental group. Experimental animals received intravenous injection of alloxan hydrochloride (Sigma Chemical Co., St. Louis, Missouri) (100 mg./kg.). The development of diabetes was verified by measuring body weight and blood glucose levels. After 12 weeks clitoral cavernous blood flow in ml. per minute per 100 gm. tissue was measured with a laser Doppler flowmeter. Cross sections of the clitoris were used for histochemistry and histomorphometric image analysis. RESULTS: After 12 weeks 5 animals were included in the diabetes group. Mean baseline flaccid and peak clitoral cavernous blood flow plus or minus standard deviation significantly decreased in the diabetic group compared with the control group (3.9 +/- 1.6 and 5.8 +/- 2.2 versus 7.2 +/- 2.5 and 12.9 +/- 5.8 ml. per minute per 100 gm. tissue, respectively, p <0.05). Histology revealed diffuse clitoral fibrosis in the diabetic group. On histomorphometry the mean proportion of clitoral cavernous smooth muscle in the diabetic group was significantly decreased compared with the control group (51.9% +/- 4.9% versus 62.3% +/- 3.1%, p < 0.05). CONCLUSIONS: These results show that diabetes mellitus produces significant adverse effects on the hemodynamic mechanism of clitoral engorgement and leads to diffuse clitoral cavernous fibrosis. It implies that decreased sexual arousal in diabetic women may result from structural changes in the clitoris.

Cisplatin decreases the abundance of aquaporin water channels in rat kidney.

The present study examined whether the cisplatin induced urinary concentration defect can be related to an altered regulation of aquaporin (AQP) water channels in the kidney. Cisplatin (8 mg/kg) was injected intraperitoneally into male Sprague-Dawley rats. The control group was without cisplatin treatment. Four d later, the expression of AQP1, AQP2, and AQP3 proteins was determined in the kidney. To specify further the primary point of derangement in the pathway that activates the arginine vasopressin-mediated AQP channels, different components of adenylyl cyclase complex were examined separately. The cisplatin treatment caused a polyuric renal failure in association with decreases of free water reabsorption. The expression of AQP1 and AQP2 was decreased in the cortex, the outer medulla, and the inner medulla, whereas that of AQP3 was decreased in the outer medulla and the inner medulla. The expression of AQP2 proteins in the apical membrane-enriched fraction decreased in parallel with that in the subapical vesicle-enriched fraction, indicating a preserved targeting. Immunohistochemistry of the outer medulla also revealed that cisplatin decreased immunoreactivity for AQP1, AQP2, and AQP3. The arginine vasopressin-evoked generation of cyclic adenosine monophosphate was attenuated by cisplatin, being most prominent in the outer medulla. However, the cyclic adenosine monophosphate generation in response to forskolin was not affected, whereas that to sodium fluoride was diminished significantly. Cisplatin also decreased the expression of Gsalpha proteins in the outer medulla and the inner medulla. These results suggest that a reduced expression of AQP water channels accounts at least in part for the cisplatin-induced urinary concentration defect.