Comparison of peripapillary vascular density as measured by optical coherence tomography angiography between various types of open-angle glaucoma.

PURPOSE: The purpose of this study is to compare the peripapillary vascular density in patients with various types of open-angle glaucoma (primary open-angle glaucoma, pseudoexfoliative glaucoma and pigmentary glaucoma) with healthy subjects by optic coherence tomography angiography (OCTA). METHODS: Twenty-seven eyes with diagnosed POAG, thirty-four eyes with diagnosed PXG, twenty eyes with diagnosed PG and thirty eyes of healthy individuals were included in our study. Peripapillary vessel density measurements were performed with all images; (AI-DD), intra-disc (ID-DD) and peripapillary (PP-DD); measurement of vascular density in the radial peripapillary capillary network was performed by OCTA. The Kruskal-Wallis test and post-hoc test were performed for statistical analysis. RESULTS: AI-DD was 50.76±2.07% in the healthy group, 47.12±2.57% in POAG, 39.71±6.64% in PXG, and 43.37±1.55% in PG. ID-DD was 50.49±3.74% in the healthy group, 49.51±6.83% in POAG, 38.42±13.46% in PXG, and 40.9±4.45% in PG. PP-DD was 51.26±3.12% in the healthy group, 50.13±3.04% in POAG, 42.31±7.31% in PXG, and 47.6±1.40% in PG. While it was found that all image and intra-disc vascular density measurements were statistically significantly lower in the PG and PXG group compared to the healthy group and the POAG (P<0.001), there was no significant difference between PXG and PG or between the control group and POAG. CONCLUSIONS: The lower radial peripapillary capillary density in PXG and PG compared to the POAG and healthy groups suggests that the blood flow around the optic disc is negatively affected in these patients.

Continuous and intermittent parathyroid hormone administration promotes osteogenic differentiation and activity of programmable cells of monocytic origin.

Bone healing deficiencies are challenging for orthopedic practice. The use of stem cells with scaffolds to treat bone tissue losses currently is popular for promoting regeneration of tissue. Programmable cells of monocytic origin (PCMO) may differentiate into three germ layers and may be a promising alternative treatment due to their stem cell-like properties. Parathyroid hormone (PTH) participates in bone metabolism. Intermittent administration of PTH promotes osteogenic activity of mesenchymal stem cdells (MSC). We investigated the osteogenic effects of continuous and intermittent administration of PTH on PCMO. Mononuclear cells were harvested from the peripheral blood of healthy donors. Isolated cells were cultured for six days in a de-differentiation medium. Indirect immunocytochemistry using anti-CD14, anti-CD45 and anti-CD90 primary antibodies, as well as electron microscopy were used to detect PCMO. PCMO then were cultured in an osteogenic differentiation medium supplemented with continuous or intermittent 50 ng/ml PTH. The PTH-free control group (CG), intermittent PTH treated group (IPG) and continuous PTH treated group (CPG) were cultured and assessed for their differentiation into osteogenic lineage cells by indirect immunocytochemistry using anti-collagen I, anti-osteonectin and anti-osteocalcin primary antibodies. Osteoblast-like cells obtained by continuous or intermittent PTH administration exhibited increased levels of collagen I, osteonectin and osteocalcin immunoreactivity. We found that continuous and intermittent PTH administration to PCMO enhanced their differentiation to osteogenic lineage cells and increased osteoblastic activity.

Investigation of the effects of rapamycin on the mTOR pathway and apoptosis in metastatic and non-metastatic human breast cancer cell lines.

AIM: The aim of this study was to analyze the effects of rapamycin treatment on apoptosis via mTOR pathway in metastatic and non-metastatic human breast cancer cell lines by immunohistochemical and TUNEL analysis. METHOD: MCF-7 and MDA-MB 231 cell lines were incubated under standard conditions forming Rapamycin and control groups. In immunohistochemical evaluation; mTOR pathway was evaluated with anti-IGF1, anti-PI3K, anti-pAKT1/2/3, anti-mTORC1, anti-mTORC2 and anti-ERK1 antibodies. The effect of apoptosis was also confirmed by TUNEL method. RESULTS: In this study, activation of PI3K/AKT/mTOR and related molecular pathways in the MDA-MB 231 and MCF-7 breast cancer cell line was evaluated and it was observed that these pathways could play a key role in cancer development. Increased apoptotic cells were observed in mTORC1 inhibition by Rapamycin administration. CONCLUSION: Targeting the mTOR pathway in breast cancer treatment may be a treatment option. In addition, the demonstration and confirmation of increased apoptosis in Rapamycin treated groups suggested that Rapamycin, an inhibitor of mTOR, is promising in the treatment of breast cancer (Tab. 2, Fig. 3, Ref. 66).

Effects of caffeic acid phenethyl ester (CAPE) on angiogenesis, apoptosis and oxidative stress in various cancer cell lines.

Cancer is a common cause of death worldwide. Approximately 80% of cancer patients use complementary or alternative medicines for treatment. Caffeic acid phenethyl ester (CAPE), the main active component of propolis, exhibits cytotoxic, antiproliferative and anti-cancer effects. Despite its anticancer effects CAPE exhibits no known harmful effects toward normal cells. We investigated the effects of CAPE on angiogenesis, apoptosis and oxidative stress using MDA MB-231, N2a and COLO 320 cell lines and CAPE treatments at 24 and 48 h. A two dimensional cell culture system was used and the findings were evaluated by an indirect immunohistochemical method and H-scores were calculated. CAPE was effective for all three cancer cell lines. After 24 and 48 h, we found a significant decrease in live cells and increased stress in the cells based on e-NOS and i-NOS levels.

Analysis of corneal astigmatism with NIDEK axial length scan in caucasian cataract surgery candidates.

AIMS AND OBJECTIVES: The aim of the study was to analyze and quantify the pattern of corneal astigmatism in Caucasian cataract surgery patients using a new optical biometer (axial length [AL] Scan, NIDEK Co., Gamagori, Japan). PATIENTS AND METHODS: The datasets of cataract surgery patients acquired between March 1, 2014, and April 15, 2016, were collected and analyzed. The corneal power (flat keratometry, steep keratometry, and mean keratometry), negative cylinder power, and axis of astigmatism were recorded. Keratometry values were optically measured by optical low coherence interferometry (AL-Scan, NIDEK Co., Ltd.,) before cataract extraction. RESULTS: The study comprised 1233 eyes of 838 consecutive cataract candidates with a mean age of 66.8 ± 10.7 years (range 40-97 years). The mean keratometry value and corneal astigmatism were 43.69 ± 1.61 D and 0.84 ± 0.70 D, respectively. Corneal astigmatism of 1.00 D or greater was found in 344 eyes (27.9%), and 548 eyes (44.4%) had against-the-rule astigmatism. A trend toward decreasing J0 and J45 with age was found by linear regression models. The per-year increase in age was associated with a J0 and J45 decrease of 0.002 D and 0.001D, respectively. CONCLUSION: This study provides the distribution of astigmatism axis and power for cataract patients in age subsets from Turkey.

Biocompatibility of vertically aligned multi-walled carbon nanotube scaffolds for human breast cancer cell line MDA-MB-231.

The aim of the current study was to determine whether the MWCNT-based scaffold has a suitable structure for cell growth and provides a biocompatible environment for human MDA-MB-231 cell lines. MWCNT-based nanostructured scaffolds were produced by plasma-enhanced chemical vapor deposition (PECVD) technique. MDA-MB-231 cells were seeded on MWCNTs-textured silicon scaffolds and on pristine silicon surfaces. After 1 week of culturing, the scaffolds were prepared for SEM analysis and immunocytochemical staining was performed for the two groups (MWCNT scaffold and pristine silicon surface), using MMP-2, MMP-9, PI3K, AKT and NF-κB primary antibodies. SEM analyses showed that the MDA-MB-231 cells better adhered to the MWCNT-based nanostructured scaffold than the pristine silicon surface. Immunohistochemical activity of the MDA-MB-231 cells on both materials has similar staining with anti-AKT MMP-2, MMP-9 and NF-κB primary antibodies. Therefore, the results of the present study suggest that the MWCNT-based scaffolds enhanced cell adhesion to the scaffold and exhibited more biomimetic properties and physiological adaptation with the potential to be used for in vitro metastasis studies of BrCa cell lines.

Assessment of mTOR pathway molecules during implantation in rats.

Mammalian target of rapamycin (mTOR) is a member of the PI3K/Akt/mTOR signaling pathway that participates in cell growth, proliferation, protein synthesis, transcription, angiogenesis, apoptosis and autophagy. We investigated the role of mTOR and other signaling molecules in the rat uterus during implantation. Female pregnant rats were divided into three groups: embryonic days (ED) 4.5, 5.5 and 6.5 according to vaginal smears. Immunohistochemical staining of mTORC1, mTORC2, IGF1, PI3K, pAkt1/2/3, ERK1 and pERK1/2 was performed on formalin fixed, paraffin embedded uterine tissue samples. pAkt1/2/3 and ERK1 also were analyzed using western blotting. We found that PI3K/Akt/mTOR and ERK/pERK were increased during the implantation period. Different amounts of mTORC1, mTORC2, IGF1, PI3K, pAKT1/2/3, ERK1 and pERK1/2 were expressed in luminal epithelium, decidual cells, embryoblast and trophoblast cells during implantation. We suggest that mTOR and associated signaling molecules may participate in implantation.

The effect of local application of insulin-like growth factor for prevention of inner-ear damage caused by electrode trauma.

BACKGROUND: Electrode insertion during cochlear implantation causes cochlear damage and apoptosis. Insulin-like growth factor applied locally was investigated in 21 rats. METHODS: In the sham group, an intracochlear dummy electrode was inserted through the round window. In the control group, after the same insertion procedure, saline-soaked porcine skin gelatine was placed on the round window. In the study group, insulin-like growth factor 1 soaked gelatine was placed on the round window. Auditory brainstem response thresholds were measured and histopathological examination was performed. RESULTS: In the study group, at 2-4 kHz, one rat had deterioration, one showed improvement and the rest had stable thresholds 14 days after intervention. At 6 kHz, four rats showed improvement and the rest remained stable. At 8 kHz, four showed improvement, one had deterioration and two remained stable. In the other groups, hearing loss deteriorated in about half of the rats and remained stable in the rest. The mean post-operative 6 kHz threshold was significantly lower than that immediately after the intervention in the study group, contrary to the other groups. The study group had significantly better mean histopathological grading than the other groups. CONCLUSION: Local insulin-like growth factor 1 application may protect hearing after cochlear implantation.

Effects of high doses of dexamethasone on hemodynamic and immunohistochemical characteristics of acute paraquat intoxication in rat kidneys.

Paraquat (1,1'-dimethyl-4,4'-bipyridinium) (PQ), is a nonselective contact herbicide that is highly toxic to humans. The kidney is affected during PQ intoxication. Dexamethasone (Dexa) has anti-inflammatory effects and is used to treat cases of PQ poisoning. We investigated in rat kidney hemodynamic effects and immunohistochemical characteristics of Dexa treatment in acute PQ poisoning. Adult male rats were divided into four groups: 1, untreated control; 2, treated with 100 mg/kg Dexa; 3, treated with 25 mg/kg PQ; 4, treated with PQ + Dexa. Mean arterial pressure (MAP) and heart rate (HR) were recorded during the experimental period (2 h). Tissues were removed after 2 h and immunohistochemistry was performed after 24 h. Paraffin sections of kidney were prepared and anti-cyclo-oxygenase-1 (COX-1), anti-cyclo-oxygenase-2 (COX-2), anti-angiotensin converting enzyme (ACE), anti-aquaporin-1 (AQU-1), anti-vascular cell adhesion molecule (VCAM) primary antibodies were used for immunohistochemical examination. Immunoreactivities were scored as: (1) minimal, (2) weak, (3) mild, (4) moderate, (5) strong and (6) very strong. MAP and HR were measured at 10 min, 20 min, 1 h and 2 h. MAP at 10 and 20 min and 1 h was increased in the Dexa group. HR also was increased in all groups compared to controls at 2 h. Compared to groups 2 and 4, MAP values decreased significantly in group 3 at 1 h. The intensity of all of immunoreactivities was decreased in group 2. In group 3, immunoreactivities of COX-1, COX-2 and ACE were decreased compared to the control and the other groups, whereas AQU-1 and VCAM immunoreactivities were the same as the control group. ACE and VCAM immunoreactivities were decreased in group 4 compared to the control group, while COX-1, COX-2 and AQU-1 immunoreactivities were close to those of the control group. Dexa appears to be useful for treating PQ intoxication.

The expression of Forkhead transcription factors in decidua and placenta in patients with missed abortion.

BACKGROUND: Forkhead transcription factors 3a (FOXO3a) has pleiotropic biological functions in the female reproductive tract. FOXO3a has a function in decidualization, in placental development, and also in inhibition of apoptosis. This study aims to investigate a possible role of FOXO3a in missed abortion. MATERIALS AND METHODS: Decidual and placental tissue samples were obtained from the women with unwanted pregnancy as the control group and with missed abortion as the patient group. Immunohistochemistry technique was utilized to compare FOXO3a expression of the decidual cells in uterine decidual stroma and cytotrophoblast-syncytiotrophoblast cells in placental villous stroma. Immunohistochemistry was evaluated semiquantitatively utilizing the H-score technique. Results: It was demonstrated that H-Scores of FOXO3a expression in both uterine decidual stroma were increased in the missed abortion group (255.83 +/- 12.41) than in the normal pregnancy group (133.33 +/- 17.43). It was also shown that there was no difference between non-decidual area of the endometrium of the normal pregnancy and the missed abortion group (30.33 +/- 4.32; 39.66 +/- 14.30, respectively) and placental villous stroma (13.00 +/- 1.89; 13.00 +/- 1.67, respectively). However, the immunoreactivity of cytotrophoblast and syncytiotrophoblast cells significantly increased in the missed abortion group (18.83+1.47; 322.00 +/- 6.06, respectively) than in the normal pregnancy group (11.00 +/- 1.26; 254.00 +/- 8.17, respectively) (p < 0.05). CONCLUSION: These data support the hypothesis that increased FOXO3a expression in missed abortion may prevent the discharge of dead fetus to maintain decidualization, prevention of oxidative stress, immunomodulation, and inhibition of apoptosis.

The role of hypoxia related angiogenesis in uterine smooth muscle tumors.

Mechanisms of hypoxia-related angiogenesis are important for uterine smooth muscle tumors. Factors that are related to angiogenesis during hypoxia include vascular endothelial growth factor (VEGF), hypoxia inducible factor 1α (HIF1α), T-cell intracellular antigen1 (TIA1), eukaryotic translation initiation factor 2α (eIF2α) and thrombospondin 1 (TSP1). We investigated immunoreactivities of VEGF, HIF1α, TIA1, eIF2α and TSP1 using an indirect immunoperoxidase method for formalin fixed, paraffin embedded tumors that had been diagnosed as leiomyoma (LMY), cellular leiomyoma (CLM) or leiomyosarcoma (LMS). TSP1 immunoreactivity was scored as moderate, mild or minimal, while VEGF, eIF2α and TIA1 immunoreactivities were scored as mild, moderate and strong in LMY, CLM and LMS samples, respectively. HIF1α immunoreactivity was scored as mild to minimal in LMY, CLM and LMS samples, but showed no statistically significant differences among samples. Although angiogenic factors showed strong immunohistochemical staining intensity in LMS, anti-angiogenic factors showed minimal immunohistochemical intensity. There was no difference in HIF-1α immunoreactivity compared to LMY, CLM and LMS samples. We suggest that HIF1α protein synthesis could be suppressed by eIF2α and TIA1. Furthermore, VEGF could be activated by pathways such as COX2, Ras, NF-ĸB or c-myc instead of HIF1α. Angiogenesis could trigger and accelerate tumor development; therefore, anti-angiogenic therapy could be useful for treatment of tumors.

Histopathological and audiological effects of mechanical trauma associated with the placement of an intracochlear electrode, and the benefit of corticosteroid infusion: prospective animal study.

OBJECTIVE: This study aimed to present the histopathological and audiological effects of mechanical trauma associated with the placement of a model electrode in the scala tympani in rats, and the effects of continuous topical corticosteroid application. METHOD: The study comprised three groups of rats. The round window membrane was perforated in all three groups and a model electrode was inserted in the round window. Group one received no further treatments. Groups two and three also had an intrathecal microcatheter compatible with a mini-osmotic pump inserted; in group two this was used to release normal saline and in group three the pump released 400 µg/ml dexamethasone. RESULTS: Dexamethasone infusion given after implantation of the intracochlear model electrode was more effective for preventing hearing loss than the administration of just one dose of dexamethasone. CONCLUSION: The findings suggest that continuous dexamethasone infusion is beneficial for preventing the loss of hair cells and neurons associated with early and late periods of intracochlear electrode trauma.

Effects of acute treatment with dexamethasone on hemodynamic and histopathological changes in rats.

We assessed the time-dependent effects of intraperitoneal (i.p.) and intravenous (i.v.) application of dexamethasone (Dexa) on the mean arterial blood pressure (MAP), heart rate (HR) and total blood volume (TBV). We evaluated also the relation between the effects and immunoreactivities of transforming growth factor-beta (TGF-ß), epithelial nitric oxide synthase (eNOS), interleukin-1 beta (IL1-ß) and vascular endothelial growth factor (VEGF) in rat brain, lung and kidney tissues. Rats were anesthetized and while still breathing spontaneously, a tracheotomy and femoral vein and artery catheterizations were performed. To determine TBV using the hemodilution method, 2 ml albumin-electrolyte solutions were applied by i.v. injection. Group 1 (control group) received a 1 ml bolus injection of physiologic saline, Group 2 received 15 mg/kg and Group 3 received 75 mg/kg Dexa i.p. The hematocrit was measured at 10, 20, 60 and 120 min. For each animal, the values of MAP, HR and TBV were measured within 2 h. For immunohistochemical evaluation, anti-TGF-ß, anti-eNOS, anti-IL1-ß and anti-VEGF primary antibodies were tested using the avidin-biotin-peroxidase method. TBV was decreased in Group 1 and the increase in MAP was statistically significant. HR values increased slightly. None of the values changed significantly in Group 2. Although TBV was unchanged in Group 3, the decrease in MAP was statistically significant. HR values increased, but the increase was not statistically significant. Mild IL1-ß immunoreactivity and moderate TGF-ß, eNOS and VEGF immunoreactivities were observed in the brain, lung and kidney samples in Group 1. Increased eNOS immunoreactivity in the kidney samples were observed in Group 2. eNOS immunoreactivity was as strong in the brain and the kidney samples in Group 3. Decreased VEGF immunoreactivity was observed in the lung and kidney tissues in Group 3. Significantly decreased TGF-ß immunoreactivity was observed in all tissue samples in Group 3. The decreased MAP values in Group 3 differed from those in Groups 1 and 2. Despite increased eNOS immunoreactivity, especially in brain and kidney, the decrease in VEGF immunoreactivity in Group 3, especially lung and kidney, were consistent with a drop in blood pressure.

Matrix metalloproteinase (MMP-2 and MMP-9) and steroid receptor expressions in feline mammary tumors.

We evaluated the presence of estrogen (ER) and progesterone (PR) receptors, and matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) enzymes in 18 feline mammary tubulopapillary carcinomas. Immunohistochemistry was performed to localize ER, PR, MMP-2 and MMP-9 in situ. Western blotting and zymographic analyses also were performed to investigate the presence and activities of MMP-2 and MMP-9 enzymes in fresh tissue homogenates. ER immune expression was detected in five samples (27.7%) and PR was positive in sixteen (88.8%) samples. Diffuse cytoplasmic staining of MMP-2 and MMP-9 in neoplastic mammary epithelial cells, stromal fibroblasts and inflammatory cell was evident. MMP-2 and MMP-9 staining was observed also in metastasizing neoplastic cells within lymphatic vessels. MMP-2 and MMP-9 enzymes and their activities in fresh tumor homogenates were demonstrated by zymography. Comparison of MMP-9 gelatinolytic bands from tumor samples and controls revealed a statistically significant difference. We demonstrated elevated MMP-9 and MMP-2 levels in tumor samples by Western blotting; analysis of protein bands revealed 1.9-to-3 fold increase in MMP-9 in tumor samples and the difference was statistically significant. Our results suggest that the expression of MMP-9 can be an important indicator for tumor progression and the possible metastatic nature of feline tubulopapillary carcinomas.

Effect of GnRH antagonist therapy on the expression of MUC-1 and heparin binding growth factor expression in the endometrium of hyperstimulated rats.

PURPOSE: To determine the effect of GnRH-antagonist therapy on the expression of heparin binding-epidermal growth factor (HB-EGF) and MUC-1 glycoprotein in hyperstimulated rat ovaries. METHODS: 30 female Wistar rats were divided into three groups (control, FSH and FSH+cetrorelix). Control rats were given 0.2 ml oil/saline mixture for four days beginning from the day of estrus. In the second group, 30 IU/ml purified hFSH was injected SC for four days beginning from the day of estrus. The rats of the third group were injected 30 IU FSH for four days and 10 IU cetrorelix SC for three or four days. The rats were sacrificed and the staining intensity of HB-EGF and MUC-1 of the epithelial cells and stromal cells of the endometrium of the rats was calculated by H-score. RESULTS: Slight MUC-1 immunoreactivity was seen in the epithelial and decidual cells of the control and FSH groups. In the FSH+cetrorelix group, moderate MUC-1 immunostaining appeared in the epithelial and desidual cells. In rats in the control and FSH+cetrorelix groups, HB-EGF immunoreactivity in the epithelial cells and decidual cells was moderate. Strong immunoreactivity was seen in the FSH group. When the MUC-1 H-score values were compared statistically with the control and other groups, FSH+cetrorelix immunoreactivity in epithelial and decidual cells were significantly different from control and FSH groups. HB-EGF immunoreactivity of the epithelium and decidua was similar in the control and FSH+cetrorelix groups, but epithelial and decidual immunoreactivity of the FSH group was different from the other two groups. CONCLUSION: Our findings suggest that GnRH antagonists exert direct effects on the expression of HB-EGF and MUC-1 expression in the rat endometrium.

A case of schistosoma reflexum in a cat with chromosomal aberrations.

A 2-year-old, female Persian cat was presented with a history of distocia. In her first pregnancy, she had whelped four kittens and had eaten all of them right after parturition. She had mated again with the same tomcat. Well-developed foetuses with weak foetal heart beats were observed in the ultrasonographic examination. En block ovariohysterectomy was performed. Three live and mature foetuses were obtained from the uterus; two of them were female foetuses and had no anatomical problem but the third one exhibiting multiple malformations was a male and diagnosed as 'schistosoma reflexum' (SR). The vertebral column deviated markedly to the right (scoliosis) at thoracolumbar region, and the middle lumbar and the sacral vertebrae were directed dorsocranially (lordosis). The entire small intestine, a part of large intestine, stomach, spleen and the right kidney were displayed out of the body, and it seemed that the listed internal organs were protruded from an abdominal cleft associated with the allantoic membrane. Liver, lungs and heart were hypoplastic. The large intestine was seen to have blind end (atresia recti), but anus was normal. Cerebrum and cerebellum were noticed as normal in sizes. Chromosome preparations from lymphocyte cultures of the foetus showed chromosomal aberrations including chromatid and chromosome breaks, exchange figures, non-homologous pairing, whereas no abnormalities were detected in the chromosome preparations from mother's cultures. This is probably the first case of SR in a cat, which was examined in detail from clinical, pathological, radiological and chromosomal angles.

Nalfurafine prevents 5'-guanidinonaltrindole- and compound 48/80-induced spinal c-fos expression and attenuates 5'-guanidinonaltrindole-elicited scratching behavior in mice.

The aims of the present study were to establish if nalfurafine, a kappa opioid agonist, inhibits compulsive scratching in mice elicited by the s.c. administration (behind the neck) of 5'-guanidinonaltrindole (GNTI), a kappa opioid antagonist; to assess if nalfurafine prevents c-fos expression provoked by GNTI or compound 48/80, two chemically diverse pruritogens; and to distinguish on the basis of neuroanatomy, those neurons in the brainstem activated by either GNTI-induced itch or formalin-induced pain (both compounds given s.c. to the right cheek). Pretreatment of mice with nalfurafine (0.001-0.03 mg/kg s.c.) attenuated GNTI (0.3 mg/kg)-evoked scratching dose-dependently. A standard antiscratch dose of nalfurafine (0.02 mg/kg) had no marked effect on the spontaneous locomotion of mice. Tolerance did not develop to the antiscratch activity of nalfurafine. Both GNTI and compound 48/80 provoked c-fos expression on the lateral side of the superficial layer of the dorsal horn of the cervical spinal cord and pretreating mice with nalfurafine inhibited c-fos expression induced by both pruritogens. In contrast to formalin, GNTI did not induce c-fos expression in the trigeminal nucleus suggesting that pain and itch sensations are projected differently along the sensory trigeminal pathway. Our data indicate that the kappa opioid system is involved, at least in part, in the pathogenesis of itch; and that nalfurafine attenuates excessive scratching and prevents scratch-induced neuronal activity at the spinal level. On the basis of our results, nalfurafine holds promise as a potentially useful antipruritic in human conditions involving itch.

The effects of DBcAMP on the expression of eNOS, iNOS and TGF-beta in rat heart tissue.

Dibutryl (DB) adenosine 3',5'-cyclic monophosphate (cAMP) is an important modulator of physiological functions. To determine the protective effects of DBcAMP on heart tissue, we evaluated changes in immunoreactivity of endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS) and transforming growth factor-beta (TGF-beta) in left cervical vagotomized rats treated with DBcAMP. Male rats were divided into four groups. In Group 1, animals were subjected to a left cervical vagotomy. Group 2 received a 1 ml bolus injection of 15 ml/kg DBcAMP in addition to the left vagotomy. DBcAMP alone was given to Group 3 and Group 4 was the control group. For each animal, mean arterial pressure (MAP) and heart rate (HR) were measured. For indirect immunohistochemistry, anti-eNOS, anti-iNOS, and anti-TGF-beta primary antibodies were used. In Group 1, MAP and HR values decreased slightly. In Groups 2 and 3, DBcAMP induced a statistically significant drop in HR and MAP. In Group 1, strong eNOS, iNOS, and TGF-beta immunoreactivities were observed. Immunostaining intensities decreased in Groups 2 and 3. The results of the study reported here suggest that increased immunoreactivities of eNOS, iNOS, and TGF-beta might contribute to the effects on the heart tissue after left vagotomy and imply that DBcAMP acts on heart tissue via nitric oxide.

Comparison of the diuretic effects of chemically diverse kappa opioid agonists in rats: nalfurafine, U50,488H, and salvinorin A.

Kappa opioid receptor agonists induce water diuresis in animals and humans. We investigated the effects of s.c. nalfurafine, U50,488H, salvinorin A, and its longer-acting analog, 2-methoxymethyl-salvinorin B (MOM-sal B), on urinary output and sodium excretion over 5 h in euvolemic rats. Nalfurafine (0.005-0.02 mg/kg), U50,488H (0.1-10 mg/kg), and MOM-sal B (0.625-5 mg/kg) induced diuresis dose-dependently. Systemically (0.1-10 mg/kg) or centrally (50 microg, i.c.v.) administered salvinorin A was ineffective. 5'-Guanidinonaltrindole, a kappa receptor antagonist, inhibited nalfurafine- and MOM-sal B-induced diuresis. Nalfurafine and MOM-sal B had no effect on arginine vasopressin levels, measured at 2 h. Tolerance did not develop to the diuresis accompanying subchronic administration of nalfurafine (0.02 mg/kg). On the basis of our work, we (a) promote nalfurafine as a candidate diuretic to relieve water retention and (b) highlight salvinorin A as a kappa agonist that does not cause diuresis, probably because of its short duration of action.

Immunolocalizations of VEGF, its receptors flt-1, KDR and TGF-beta's in epithelial ovarian tumors.

OBJECTIVE: Angiogenesis is an essential factor for growth, differentiation, invasion and metastasis of tumors. In this study, we aimed to evaluate the immunolocalizations of vascular endothelial growth factor (VEGF), its receptors flt-1, KDR/flk-1, and transforming growth factor-beta's (TGF-beta) in epithelial ovarian tumors, utilizing indirect immunohistochemistry to understand the role of the angiogenic events in ovarian neoplasia. METHODS: Tissue blocks from 40 patients who had ovarian pathology (borderline serous-mucinous tumor and malignant serous-mucinous adenocarcinoma of the ovary) were included in this study. All formalin-fixed, paraffin-embedded tissue sections were stained with hematoxylin-eosin or primary antibodies against VEGF, flt-1, KDR/flk-1, TGF-beta1, TGF-beta2 and TGF-beta3 using the avidin-biotin-peroxidase method. H-SCORE, a semi-quantitative grading system, was used to compare immunohistochemical staining intensities. RESULTS: Positive VEGF immunoreactivity was concentrated in the epithelial and stromal parts of all the ovarian samples and the endothelial cells in the stroma were also stained. Increased immunoreactivity of VEGF was observed in malignant ovarian adenocarcinomas compared to the borderline tumors of the ovary. VEGF receptors, flt-1 and KDR/flk-1 immunoreactivities were detected not only in vascular endothelial cells, but also in tumor cells at malignant sites. Immunoreactivities of VEGF and its receptors were coexpressed in tumor cells of the ovarian carcinoma. While immunoreactivities of TGF-beta1 and TGF-beta2 were both overexpressed in malignant ovarian carcinomas, immunoreactivity of TGF-beta3 was still mild. CONCLUSION: Our results suggest that overexpression of VEGF, its receptors flt-1, KDR/flk-1 and TGF-beta interaction may play an important role in the ovarian cancer biology, with potential effects on tumor growth and angiogenesis. New therapeutic strategies using VEGF and TGF-beta antagonists could obtain an additional approach to the treatment ovarian carcinoma by inhibiting angiogenesis.