Sonochemical synthesis of graphene oxide-Ag2O nanozyme as an oxidize-like mimic for the highly sensitive detection of lithium in blood serum.

Bipolar disorder is commonly treated with lithium carbonate. The concentration of lithium in the blood serum should be closely monitored in patients who require long-term lithium therapy. To date, no colorimetric method of detecting lithium ions has been reported using nanosensors. We have developed a novel chemosensor based on nanozyme (NZ) to address this clinical need. The GO-Ag2O NZs were synthesized by a sonochemical method and used as a colorimetric nanosensor to detect lithium ions in human blood serum (Li (I)). To characterize NZs, various techniques were employed, including XRD, FTIR, TEM, FESEM, EDX, Raman spectroscopy, BET, DLS, Zeta potential, and ICP-OES. According to TEM and FESEM images of GO-Ag2O, the nanoparticles (NPs) of Ag2O are uniformly distributed on the surface of 2D graphene oxide sheets. In addition, silver oxide nanoparticles exhibited a cubic morphology with an average size of 3.5 nm. We have examined the performance of the NZs in an aqueous medium and in human blood serum that contains Li (I). A colorimetric test revealed that NZs synthesized in the presence of ultrasound were more sensitive to Li (I). According to the linearity of the calibration curves' ranges, Li (I) has a limit of detection (LOD) of 0.01 µg/mL. Furthermore, it displayed a linear range between 0 and 12 µg/mL. GO-Ag2O NZs showed noticeable color changes from green to orange after exposure to Li (I). An incubation time of two minutes was found to be the most effective for sensing. This innovative approach provides a reliable method for monitoring lithium levels and ensuring patient safety during long-term lithium therapy for bipolar disorder.

Optical Coherence Tomography Angiography Findings after Acute Intraocular Pressure Elevation in Patients with Diabetes Mellitus versus Healthy Subjects.

Purpose: To assess the changes in optic nerve head and macular microvascular networks after acute intraocular pressure (IOP) rise in healthy eyes versus the eyes of diabetic patients. Methods: In this prospective, interventional, comparative study, 24 eyes of 24 adults including 12 eyes of healthy nondiabetic subjects and 12 eyes with mild or moderate non-proliferative diabetic retinopathy (NPDR) were enrolled. IOP elevation was induced by a suction cup attached to the conjunctiva. IOP and optical coherence tomography angiographic (OCTA) images of the optic disc and macula were obtained before and immediately after the IOP rise. Results: Baseline and post-suction IOPs were not significantly different between the two groups (all P > 0.05). The mean IOP elevation was 13.93 ± 3.41 mmHg among all eyes and was statistically significant as compared to the baseline in both groups (both P < 0.05). After IOP elevation, healthy eyes demonstrated a reduction in the vessel density in the whole image deep and superficial capillary plexuses and parafoveal deep capillary plexus (DCP) (all P < 0.05). In diabetic retinopathy, foveal vessel density at DCP decreased significantly following IOP rise (P = 0.003). In both groups, inside disc vessel density decreased significantly after IOP rise (both P < 0.05), however, no significant change was observed in peripapillary vessel density (both P > 0.05). Conclusion: Acute rise of IOP may induce different levels of microvascular changes in healthy and diabetic eyes. Optic disc microvasculature originating from the posterior ciliary artery may be more susceptible to IOP elevation than that of retinal microvasculature.

Nobiletin prevents amyloid ß1-40-induced cognitive impairment via inhibition of neuroinflammation and oxidative/nitrosative stress.

Alzheimer's disease (AD) is presented as an age-related neurodegenerative disease with multiple cognitive deficits and amyloid ß (Aß) accumulation is the most important involved factor in its development. Nobiletin is a bioflavonoid isolated from citrus fruits peels with anti-inflammatory and anti-oxidative activity as well as anti-dementia property that has shown potency to ameliorate intracellular and extracellular Ab. The aim of the present study was to assess protective effect of nobiletin against Aß1-40-induced cognitive impairment as a consistent model of AD. After bilateral intrahippocampal (CA1 subfield) injection of Aß1-40, rats were treated with nobiletin (10 mg/kg/day; p.o.) from stereotaxic surgery day (day 0) till day + 7. Cognition function was evaluated in a battery of behavioral tasks at week 3 with final assessment of hippocampal oxidative stress and inflammation besides Nissl staining and 3-nitrotyrosine (3-NT) immunohistochemistry. Analysis of behavioral data showed notable and significant improvement of alternation in Y maze test, discrimination ratio in novel object recognition task, and step through latency in passive avoidance test in nobiletin-treated Aß group. Additionally, nobiletin treatment was associated with lower hippocampal levels of MDA and ROS and partial reversal of SOD activity and also improvement of Nrf2 with no significant effect on GSH and catalase. Furthermore, nobiletin attenuated hippocampal neuroinflammation in Aß group as shown by lower tissue levels of TLR4, NF-kB, and TNFa. Histochemical findings showed that nobiletin prevents CA1 neuronal loss in Nissl staining in addition to its alleviation of 3-nitrotyrosine (3-NT) immunoreactivity as a marker of nitrosative stress. Collectively, these findings indicated neuroprotective and anti-dementia potential of nobiletin that is partly attributed to its anti-oxidative, anti-nitrosative, and anti-inflammatory property associated with proper modulation of TLR4/NF-kB/Nrf2 pathways.

Effects of crocin and metformin on methylglyoxal-induced reproductive system dysfunction in diabetic male mice.

OBJECTIVE: This study investigated the effect of crocin in methylglyoxal (MGO)-induced diabetic male mice. METHODS: Seventy 1-month-old male NMRI mice weighing 20-25 g were divided into seven groups (n=10): sham, MGO (600 mg/kg/day), MGO+crocin (15, 30, and 60 mg/kg/day), MGO+metformin (150 mg/kg/day), and crocin (60 mg/kg/day). MGO was administered orally for 30 days. Starting on day 14, after confirming hyperglycemia, metformin and crocin were administered orally. On day 31, plasma and tissue samples were prepared for experimental assessments. RESULTS: Blood glucose and insulin levels in the MGO group were higher than those in the sham group (p<0.001), and decreased in response to metformin (p<0.001) and crocin treatment (not at all doses). Testis width and volume decreased in the MGO mice and improved in the crocin-treated mice (p<0.05), but not in the metformin group. Superoxide dismutase levels decreased in diabetic mice (p<0.05) and malondialdehyde levels increased (p<0.001). Crocin and metformin improved malondialdehyde and superoxide dismutase. Testosterone (p<0.001) and sperm count (p<0.05) decreased in the diabetic mice, and treatment with metformin and crocin recovered these variables. Luteinizing hormone levels increased in diabetic mice (p<0.001) and crocin treatment (but not metformin) attenuated this increase. Seminiferous diameter and height decreased in the diabetic mice and increased in the treatment groups. Vacuoles and ruptures were seen in diabetic testicular tissue, and crocin improved testicular morphology (p<0.01). CONCLUSION: MGO increased oxidative stress, reduced sex hormones, and induced histological problems in male reproductive organs. Crocin and metformin improved the reproductive damage caused by MGO-induced diabetes.

Awareness of Iranian Medical Sciences Students Towards Basic Life Support; a Cross-Sectional study.

INTRODUCTION: Augmentation of the number of trained basic life support (BLS) providers can remarkably reduce the number of cardiac arrest victims. The aim of this study was to evaluate the level of BLS awareness among students of medical sciences in Iran. METHODS: This multicenter cross-sectional study was performed on medical students at the 4 major medical schools in Tehran, the capital of Iran, between Jan 2018 and Feb 2019, using convenience sampling method. The level of medical sciences students' awareness of BLS was measured using an international questionnaire. RESULTS: Finally, 1210 students with the mean age of 21.2 ± 2.3 years completed the survey (79% female). 133 (10.9%) students had CPR experience and none had received any formal training. None of the responders could answer all questions correctly. The mean awareness score of participants was 11.93 ± 2.87 (range: 10.13 -17.25). The awareness score of participants was high in 49 (4.04 %) participants, moderate in 218 (18.01%), and low in 943 (77.93%) of studied cases. CONCLUSION: Based on the findings of this study, more than 70% of the studied medical sciences students obtained a low score on BLS awareness.

Spatial distribution of diabetic capillary non-perfusion.

OBJECTIVE: To evaluate the distribution of capillary non-perfusion (CNP) in superficial and deep capillary plexuses (SCP and DCP) in eyes with diabetic retinopathy (DR). METHODS: In this retrospective case series, macular optical coherence tomography angiography (OCTA) images were obtained from eyes with DR without diabetic macular edema (DME). The area of CNP in SCP and DCP was delineated using an automated approach after excluding the foveal avascular zone (FAZ) and major retinal vessels. The distribution and spatial correlation of the CNP in each layer were analyzed. RESULTS: Forty-three eyes of 27 patients with DR with a mean age of 59.10 ± 9.05 years were included. The mean CNP area in SCP was statistically significantly higher than DCP (0.722 ± 0.437 mm2 vs. 0.184 ± 0.145 mm2 , respectively, p < .001). There was a statistically significant association between mean BCVA (0.28 ± 0.21 logMAR) and CNP area in DCP (p = .01). After automated subtraction of CNP areas in DCP from SCP, 25.43 ± 15.05% of CNP areas in the DCP had co-localized CNP areas in SCP. The CNP percentage was statistically significantly different between the concentric rings on foveal center, both in SCP and in DCP (both p < .001) showing a decreasing trend from the outer ring toward the center. CONCLUSION: In DR, SCP is more ischemic than DCP. This is in contrast to the previously described oxygenation-dependent ischemic cascade following acute retinal vascular occlusions. This study provides further insight into the retinal ischemia in DR.

Intravitreal Injection Of The Granulocyte-Colony Stimulating Factor For The Treatment Of Non-Arteritic Anterior Ischemic Optic Neuropathy: A Pilot Study.

Purpose To investigate the efficacy of intravitreal injection of granulocyte colony-stimulating factor (G-CSF) for the treatment of non-arteritic anterior ischemic optic neuropathy (NAION).Methods: Patients with acute NAION were enrolled in this prospective interventional case series. They received an intravitreal injection of 60 micrograms in 0.1 ml of G-CSF within 2 weeks of the onset of the disease. Visual acuity, visual field, intraocular pressure (IOP), corneal endothelial cell density, and peripapillary retinal nerve fiber layer (RNFL) thickness were recorded before injections and 1 week, 1 month, 3 months, 6 months, and one year after the injections. Full-field electroretinography (ERG) was obtained at the baseline, 1 month, and 12 months post- injections.Results: Fourteen eyes of 14 patients entered the study. Best-corrected visual acuity (BCVA) significantly improved in the first month following injections (p = .007), decreased subsequently, and the final BCVA showed no significant improvement (p = .278) compared to the baseline measurements. A significant decrease in RNFL thickness was observed in all quadrants compared to the baseline measurements. Also, no improvement in the visual field parameters was observed. From the toxicity aspect, no significant changes in the corneal endothelial cell density, IOP, and ERG recordings were observed.Conclusion: Intravitreal injection of G-CSF seems to be safe. The effect may last for one month and then decline.

Acute retinal necrosis: Clinical manifestation and long-term visual outcomes in a series of polymerase chain reaction-positive patients.

PURPOSE: To report the clinical spectrum, viral etiologies, therapeutic interventions, timing of rhegmatogenous retinal detachments (RRD), and visual outcomes in acute retinal necrosis (ARN) syndrome in a series of polymerase chain reaction (PCR)-positive eyes. METHODS: From January 2010 to January 2017, consecutive patients with the clinical diagnosis of ARN and a positive aqueous viral PCR were included in this observational, retrospective study. RESULTS: Nineteen eyes found to have a clinical diagnosis of ARN, of which 18 (94.7%) had a positive viral PCR. ARN was unilateral, except in one patient. None of the fellow eyes manifested ARN during follow-up. Varicella-zoster virus (VZV) was detected in 78.0% of ARN eyes. 61.1% of eyes experienced RRD. The median time for the occurrence of RRD was 12 weeks (range: 6-25 weeks) after disease onset. No correlation was found between the etiologic viral agent (VZV vs non-VZV; p = 1.000), extent of retinitis (1-2 quadrant vs 3-4 quadrants; p = 0.326), administration of intravitreal ganciclovir (injected vs not injected; p = 0.332), application of prophylactic laser retinopexy (applied vs not applied; p = 0.326), and subsequent occurrence of RRD.At a 2-year follow-up, visual impairment (VA ⩽ 20/200) and severe visual loss (VA ⩽ light perception) were significantly higher in those complicated by RRD compared to non-RRD eyes (81.8% vs 28.6%; p = 0.047, and 45.4% vs 0.0%; p = 0.004, respectively). CONCLUSION: Aqueous PCR results are highly consistent with the clinical diagnosis of ARN. Regardless of the method of management, the rate of RRD is high and is associated with a poor visual outcome.

Macular microvascular changes after intravitreal bevacizumab injection in diabetic macular edema.

OBJECTIVE: To evaluate the changes in retinal capillary plexus and the choriocapillaris after a single intravitreal injection of bevacizumab in eyes with diabetic macular edema using optical coherence tomography angiography (OCTA). DESIGN: Prospective interventional case series. PARTICIPANTS: Patients having diabetes with centre-involving diabetic macular edema. METHODS: In this prospective interventional case series, eyes with centre-involving diabetic macular edema were enrolled. Vascular density (VD), vascular diameter index (VDI), vascular length density (VLD), foveal avascular zone (FAZ) area, and foveal density (FD)-300 were measured using en face OCTA images before and 1 month after administration of intravitreal bevacizumab. VD and VDI measurements were performed in the superficial capillary plexus (SCP) and deep retinal capillary plexus (DCP) and in the choriocapillaris. Additionally, capillary nonperfusion area (CNPA) was detected automatically based on vessel distance map in 4 concentric rings around the foveal centre. The segmentation error was manually corrected, and the measurements were performed by 2 expert graders. RESULTS: Twenty-three eyes of 19 patients with a mean age of 62.76 ± 6.88 years were included. There were no significant changes in the FAZ area, FD-300, or in the VD of the foveal and parafoveal SCP and DCP. Also, VLD and VDI of the SCP and DCP remained unchanged. The change in the CNPA was not statistically significant. The VD of choriocapillaris increased significantly after injections (p = 0.005). CONCLUSIONS: FAZ area and VD of the retinal capillary plexus remained stable in the short-term period after intravitreal bevacizumab. In addition, the choriocapillaris blood flow improved after bevacizumab injection.

Differential genes expression analysis of invasive aspergillosis: a bioinformatics study based on mRNA/microRNA.

Invasive aspergillosis is a severe opportunistic infection with high mortality in immunocompromised patients. Recently, the roles of microRNAs have been taken into consideration in the immune system and inflammatory responses. Using bioinformatics approaches, we aimed to study the microRNAs related to invasive aspergillosis to understand the molecular pathways involved in the disease pathogenesis. Data were extracted from the gene expression omnibus (GEO) database. We proposed 3 differentially expressed genes; S100B, TDRD9 and TMTC1 related to pathogenesis of invasive aspergillosis. Using miRWalk 2.0 predictive tool, microRNAs that targeted the selected genes were identified. The roles of microRNAs were investigated by microRNA target prediction and molecular pathways analysis. The significance of combined expression changes in selected genes was analyzed by ROC curves study. Thirty-three microRNAs were identified as the common regulator of S100B, TDRD9 and TMTC1 genes. Several of them were previously reported in the pathogenesis of fungal infections including miR-132. Predicted microRNAs were involved in innate immune response as well as toll-like receptor signaling. Most of the microRNAs were also linked to platelet activation. The ROC chart in the combination mode of S100B/TMTC1, showed the sensitivity of 95.65 percent and the specificity of 69.23 percent. New approaches are needed for rapid and accurate detection of invasive aspergillosis. Given the pivotal signaling pathways involved, predicted microRNAs can be considered as the potential candidates of the disease diagnosis. Further investigation of the microRNAs expression changes and related pathways would lead to identifying the effective biomarkers for IA detection.

Seasonality of Acute Retinal Necrosis.

PURPOSE: To study the seasonal variability in the occurrence of acute retinal necrosis (ARN) in a series of polymerase chain reaction (PCR)-positive patients. METHODS: Consecutive patients clinically diagnosed with ARN and a positive PCR result of aqueous humor during a seven-year period were studied retrospectively. Patients' demographics, causative viral agent(s), and the date of disease onset were extracted from medical records. RESULTS: Twenty eyes of 20 patients were enrolled; the mean age at presentation was 39.6 ± 14.4 (range, 6-62) years. Nine patients were female. The most common causative agent was varicella-zoster virus in 16 patients (80%), followed by herpes simplex virus in two patients (10%). The disease onset was in winter in 10 patients (50%), and the highest incidence was in February (five patients, 25%). The cumulative occurrence of ARN was significantly higher in the first half of the year (winter and spring) compared to the second half of the year (summer and fall) (P = 0.030). In general, seasons with a high incidence of ARN were preceded by cold seasons. CONCLUSION: In our series, we observed seasonal variability in the incidence of ARN, with the highest incidence during winter and spring. However, further epidemiologic studies in different geographical areas are required to elucidate the true seasonal nature of ARN.

Effect of segmentation error correction on optical coherence tomography angiography measurements in healthy subjects and diabetic macular oedema.

PURPOSE: To evaluate the impact of segmentation error on vessel density measurements in healthy eyes and eyes with diabetic macular oedema (DMO). METHODS: In this prospective, comparative, non-interventional study, enface optical coherence tomography angiography (OCTA) images of the macula from healthy eyes and eyes with DMO were acquired. Two expert graders assessed and corrected the segmentation error. The rate of segmentation error and the changes in vessel density and inner retinal thickness after correction of the segmentation error were recorded and compared between the two groups. RESULTS: 20 eyes with DMO and 24 healthy eyes were evaluated. Intergrader agreement was excellent (intraclass correlation coefficient ≥0.9) for all parameters in both groups. The rate of segmentation error was 33% and 100% in healthy and diabetic eyes, respectively (p<0.001). Nine healthy eyes (37.5%) and all eyes with DMO (100%) were noted to exhibit a change in at least one of the foveal or parafoveal vessel density measurements. The rate of any change in foveal and parafoveal vessel densities in both the superficial and deep capillary plexus was statistically significantly higher in the diabetic group (all p<0.001). No statistically significant change was observed in mean vessel density (superficial and deep capillary plexuses) after correction of the segmentation error in healthy and DMO eyes (All p>0.05). However, the mean absolute change in the vessel density measurements was statistically significantly higher in the diabetic group (all p<0.05). The mean absolute change in superficial and deep inner retinal thickness was statistically significantly higher in DMO (p=0.02 and p=0.002, respectively). CONCLUSIONS: In this study, misidentification of retinal layers and consequent vessel density measurement error occurred in all eyes with DMO and in one-third of healthy eyes. The segmentation error should be checked and manually corrected in the OCTA vessel density measurements, especially in the presence of macular oedema.

Safety of the subconjunctival injection of doxycycline in rabbits.

Purpose: To evaluate the safety of subconjunctival injection of doxycycline in rabbit eyes. Methods: Eight white New Zealand rabbits were selected. Different concentrations of 250 micrograms (µg), 500 µg, 1000 µg, and 2000 µg in 0.1 ml were prepared for subconjunctival injection. Each concentration was injected into the two eyes of each rabbit. For each dose, dextrose was injected in one contralateral eye and the other fellow eye remained non-injected. All rabbits underwent ocular examination in the 1st, 3rd, and 30th day after injection. The rabbits were sacrificed 30 days after injections and the histopathological examination was performed. Results: No obvious change was detected in all four groups from the 1st day to the 3rd day after injection in terms of tearing, hyperaemia, and chemosis. There was no visible sign of inflammation or necrosis, and also no histological change in both clinical and histopathological examinations. Conclusion: Subconjunctival injection of doxycycline with different dosages of 250 to 2000 ug in 0.1cc in rabbit eyes was safe and no clinical or histological changes were observed after one month.

Optical coherence tomography angiography features of optic nerve head drusen and nonarteritic anterior ischemic optic neuropathy.

OBJECTIVE: To evaluate the optic disc microvasculature in optic nerve head drusen (ONHD) vasculature in comparison to acute nonarteritic anterior ischemic optic neuropathy (NAION) and normal eyes using optical coherence tomography angiography (OCT-A). METHODS: Ten eyes with ONHD, 10 eyes with acute NAION, and 10 healthy eyes were included in this prospective, comparative, observational case series. OCT-A imaging was performed on the optic discs. Qualitative grading was performed for dilation and tortuosity of the peripapillary vessels by 3 graders. Quantitative comparison was performed for peripapillary and inside disc vessel densities in nerve head (NH) and retinal peripapillary capillary (RPC) slabs. RESULTS: The intergrader agreement for dilation and tortuosity of the peripapillary vessels was poor (0.313 and 0.182 for vascular dilation in nerve head and radial peripapillary capillary enface images, respectively, and 0.478 and 0.490 for vascular tortuosity in nerve head and radial peripapillary capillary enface images, respectively). In NH en face images, the vessel density measurements were statistically significantly different between the 3 groups (all p < 0.05). In RPC en face images, the vessel density measurements were statistically significantly different between the 3 groups (all p < 0.05) except for nasal peripapillary sector (0.08). CONCLUSION: Despite poor intergrader agreement in qualitative analysis, quantitative OCT-A evaluation may differentiate optic disc edema due to NAION from pseudodisc edema due to ONHD.

TIME COURSE OF CHANGES IN OPTIC DISK NEOVASCULARIZATION AFTER A SINGLE INTRAVITREAL BEVACIZUMAB INJECTION.

PURPOSE: To evaluate the longitudinal changes in optic disk neovascularization (NVD) after intravitreal bevacizumab injection using optical coherence tomography angiography. METHODS: In this prospective, interventional, case series, eyes with NVD secondary to diabetic retinopathy were enrolled. En face optical coherence tomography angiographic images were obtained from the optic disks before and 3 hours, 6 hours, 24 hours, 7 days, and 30 days after intravitreal bevacizumab injection. The size and flow area of the neovascularization were measured by two graders. RESULTS: Eleven eyes of 9 patients with a mean age of 52.11 ± 9.48 years were included. The reduction in the NVD size and flow area was statistically significant at 24 hours, 7 days, and 30 days after injections compared with the baseline measurements (all P < 0.05). The decremental regression in the NVD size and flow area continued during the study course. The changes were not statistically significant in 3-hour and 6-hour measurements (all P > 0.05). CONCLUSION: In this study, statistically significant regression in the NVD size and flow area was observed as early as 24 hours after a single intravitreal bevacizumab injection, with a continued decreasing trend for at least a 1-month period.

Analysis of Differential Expression of microRNAs and Their Target Genes in Prostate Cancer: A Bioinformatics Study on Microarray Gene Expression Data.

Early diagnosis of prostate cancer (PCa) as the second most common cancer in men is not associated with precise and specific results. Thus, alternate methods with high specificity and sensitivity are needed for accurate and timely detection of PCa. MicroRNAs regulate the molecular pathways involved in cancer by targeting multiple genes. The aberrant expression of the microRNAs has been reported in different cancer types including PCa. In this bioinformatics study, we studied differential expression profiles of microRNAs and their target genes in four PCa gene expression omnibus (GEO) databases. PCa diagnostic biomarker candidates were investigated using bioinformatics tools for analysis of gene expression data, microRNA target prediction, pathway and GO annotation, as well as ROC curves. The results of this study revealed significant changes in the expression of 14 microRNAs and 40 relevant target genes, which ultimately composed four combination panels (miR- 375+96+663/ miR- 133b+143- 3p + 205/ C2ORF72 + ENTPD5 + GLYAT11/LAMB3 + NTNG2+TSLP) as candidate biomarkers capable to distinguish between PCa tumor samples and normal prostate tissue samples. These biomarkers may be suggested for a more accurate early diagnosis of PCa patients along with current diagnostic tests.

Aberrant expression of miR-141 and nuclear receptor small heterodimer partner in clinical samples of prostate cancer.

BACKGROUND: Prostate cancer (PCa) is the second most common cancer in men worldwide. Currently, prostate-specific antigen (PSA) test and digital rectal exam are the main screening tests used for PCa diagnosis. However, due to the low specificity of these tests, new alternative biomarkers such as deregulated RNAs and microRNAs have been implemented. OBJECTIVES: Aberrant expressions of small heterodimer partner gene (SHP, NR0B2) and mir-141 are reported in various cancers. The aim of this study was to investigate the SHP and miR-141 expression level in tissue samples of prostate cancer. METHODS: The expression level of SHP gene and miR-141 was assessed by real time PCR and their relative amounts were calculated by the Δ⁢ΔCT method. Also, IHC technique was used to determine the expression level of SHP protein. RESULTS: The miR-141 was significantly up-regulated in the samples of metastatic tumors compared to localized tumor samples (P< 0.001, 31.17-fold change). Tumor samples showed lower SHP mRNA expression levels than BPH samples (p= 0.014, 4.7-fold change). The results of paired t-test analysis showed there was no significant difference between the SHP gene expression in PCa samples and their matched tumor-adjacent normal tissue (p= 0.5). CONCLUSIONS: The data obtained in our study confirm the involvement of miR-141 in PCa progression and metastasis. These effects could be mediated by AR via down-regulation of its co-repressor protein, i.e., SHP.

Identification of Differentially Expressed K-Ras Transcript Variants in Patients With Leiomyoma.

PURPOSE: Molecular studies have demonstrated a wide range of gene expression variations in uterine leiomyoma. The rat sarcoma virus/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase (RAS/RAF/MAPK) is the crucial cellular pathway in transmitting external signals into nucleus. Deregulation of this pathway contributes to excessive cell proliferation and tumorigenesis. The present study aims to investigate the expression profile of the K-Ras transcripts in tissue samples from patients with leiomyoma. METHODS: The patients were leiomyoma cases who had no mutation in mediator complex subunit 12 ( MED12) gene. A quantitative approach has been applied to determine the difference in the expression of the 2 main K-Ras messenger RNA (mRNA) variants. The comparison between gene expression levels in leiomyoma and normal myometrium group was performed using relative expression software tool. RESULTS: The expression of K-Ras4B gene was upregulated in leiomyoma group ( P = .016), suggesting the involvement of K-Ras4B in the disease pathogenesis. Pairwise comparison of the K-Ras4B expression between each leiomyoma tissue and its matched adjacent normal myometrium revealed gene upregulation in 68% of the cases. The expression of K-Ras4A mRNA was relatively upregulated in leiomyoma group ( P = .030). In addition, the mean expression of K-Ras4A gene in leiomyoma tissues relative to normal samples was 4.475 (95% confidence interval: 0.10-20.42; standard error: 0.53-12.67). In total, 58% of the cases showed more than 2-fold increase in K-Ras4A gene expression. CONCLUSION: Our results demonstrated increased expression of both K-Ras mRNA splicing variants in leiomyoma tissue. However, the ultimate result of KRAS expression on leiomyoma development depends on the overall KRAS isoform balance and, consequently, on activated signaling pathways.

Gene expression profile of FVII and AR in primary prostate cancer.

OBJECTIVES: The ectopic expression of coagulation Factor VII has been shown in various cancers. Recently, F7 gene has been identified as a direct target of the androgen receptor in breast cancer. In this study, we examined the mRNA expression of F7 and AR in clinical sample series of prostate cancer and BPH. MATERIAL AND METHODS: All the prostate cancer patients were new cases with no medical history of surgery or chemotherapy. The tissue samples were assigned as either prostate cancer tumor (n= 45) harboring at least 80% tumor cell content, or BPH (n= 36). Relative AR and F7 mRNA expression in each tissue sample was normalized to the mean of the Ct values determined for GAPDH and PSA genes. RESULTS: Mean plasma level of prostate specific antigen (PSA) was 17.82 ± 3.71 ng/ml and 7.71 ± 1.28 ng/ml (Mean ± SEM) in PCa and BPH group, respectively. AR mean expression was up-regulated 22.468 fold in clinical tumor sample cohort (S.E., 0.175-2,916, 95% CI: 0.001-126,764, P= 0.001). The mean expression of F7 gene in tumor tissues relative to PBH samples was 6.981 (S.E., 0.099-413.001, 95% CI: 0.002-34,183, P= 0.012). ANOVA analysis of the gene expression results showed significant correlation between F7 and AR mRNA expression in tumor samples (p< 0.001). CONCLUSIONS: Our study findings suggest a link between FVII and AR in prostate cancer pathogenesis. F7 gene expression could be up-regulated via various AR mediators affecting the promoter region of the F7 gene. Should this be confirmed by further studies, it may be suggested as a potential contributing factor in prostate cancer.

NOD-scid IL2R γnull mice engrafted with human peripheral blood mononuclear cells as a model to test therapeutics targeting human signaling pathways.

BACKGROUND: Animal models of human inflammatory diseases have limited predictive quality for human clinical trials for various reasons including species specific activation mechanisms and the immunological background of the animals which markedly differs from the genetically heterogeneous and often aged patient population. OBJECTIVE: Development of an animal model allowing for testing therapeutics targeting pathways involved in the development of Atopic Dermatitis (AD) with better translatability to the patient. METHODS: NOD-scid IL2R γnull mice engrafted with human peripheral blood mononuclear cells (hPBMC) derived from patients suffering from AD and healthy volunteers were treated with IL-4 and the antagonistic IL-4 variant R121/Y124D (Pitrakinra). Levels of human (h)IgE, amount of B-, T- and plasma- cells and ratio of CD4 : CD8 positive cells served as read out for induction and inhibition of cell proliferation and hIgE secretion. Results were compared to in vitro analysis. RESULTS: hIgE secretion was induced by IL-4 and inhibited by the IL-4 antagonist Pitrakinra in vivo when formulated with methylcellulose. B-cells proliferated in response to IL-4 in vivo; the effect was abrogated by Pitrakinra. IL-4 shifted CD4 : CD8 ratios in vitro and in vivo when hPBMC derived from healthy volunteers were used. Pitrakinra reversed the effect. Human PBMC derived from patients with AD remained inert and engrafted mice reflected the individual responses observed in vitro. CONCLUSION: NOD-scid IL2R γnull mice engrafted with human PBMC reflect the immunological history of the donors and provide a complementary tool to in vitro studies. Thus, studies in this model might provide data with better translatability from bench to bedside.