In-situ forming biodegradable implants for sustained Fluocinolone acetonide release to the posterior eye: In-vitro and in-vivo investigations in rabbits.

Delivering medication to the posterior segment of the eye presents a significant challenge. Intravitreal injection has emerged as the preferred method for drug delivery to this area. However, current injectable non-biodegradable implants for fluocinolone acetonide (FA) require surgical removal after prolonged drug release, potentially affecting patient compliance. This study aimed to develop an in-situ forming biodegradable implant (ISFBI) optimal formulation containing PLGA504H and PLGA756S (50:50 w/w%) with the additive NMP solvent. The goal was to achieve slow and controlled release of FA over a two-month period with lower burst release, following a single intravitreal injection. Through morphology, rheology, stability and in-vitro release evaluations, the optimal formulation demonstrated low viscosity (0.12-1.25 Pa. s) and sustained release of FA at a rate of 0.36 µg/day from the third day up to two months. Furthermore, histopathology and in-vivo studies were conducted after intravitreal injection of the optimal formulation in rabbits' eye. Pharmacokinetic analysis demonstrated mean residence time (MRT) of 20.02 ± 0.6 days, half-life (t1/2) of 18.80 ± 0.4 days, and clearance (Cl) of 0.29 ± 0.03 ml/h for FA in the vitreous humor, indicating sustained and slow absorption of FA by the targeted retinal tissue from vitrea over the two-month period and eliminating through the anterior section of the eye, as revealed by its presence in the aqueous humor. Additionally, FA exhibited no detection in the blood and no evidence of systemic side effects or damage on the retinal layer and other organs. Based on these findings, it can be concluded that in-situ forming injectable biodegradable PLGA implants can show promise as a long-acting and controlled-release system for intraocular drug delivery.

Does Renal Mass with Venous Thrombosis always Indicate Renal Cell Carcinoma? A Case Series.

Intravascular tumor extension in the major renal veins or their tributaries, as a rare but important clinical entity that can change the disease stage, prognosis, and approach to treatment. There is limited literature on the obstruction of renal vein and IVC by tumor thrombus in other types of renal tumors that are not of RCC type. We presented four different renal tumor cases with the presence of gross renal vein or IVC thrombosis. Although the incidence of renal vein and IVC tumor thrombus might be suggestive of (often diagnosed as) RCC, the possibility of other non-RCC renal tumors should be included in the differential diagnosis.

Cardiac and renal fibrosis and oxidative stress balance in lipopolysaccharide-induced inflammation in male rats.

BACKGROUND: Subclinical inflammation induced by persistent exposure to lipopolysaccharide (LPS) is found in some clinical conditions such as obesity or diabetes. This study aimed to investigate the effect of recurrent LPS exposure on inflammatory markers, oxidative stress balance and cardiac and renal fibrosis in male rats. METHODS: Male Wistar rats were divided into control and LPS-treated. LPS (10 mg/kg/week) was injected intraperitoneally. After 4 weeks, left ventricles and kidneys were homogenized and stained with hematoxylin and eosin (H&E) and Masson trichrome for histological examination. Serum levels of nitrite, interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured and total thiol, malondialdehyde (MDA), superoxide dismutase (SOD) and catalase were evaluated in the heart and kidney homogenates. RESULTS: Serum inflammatory markers were higher in LPS group than control (nitrite: 37.0 ± 2.2 vs. 25.5 ± 1.9 µmol/l; IL-6: 84 ± 3 vs. 98.0 ± 4.4 pg/ml; TNF-α: 75.5 ± 4.9 vs. 85.3 ± 4.7 pg/ml; respectively, P < 0.050). Evaluation of total thiol concentration (heart: 10.0 ± 0.9 vs. 22.5 ± 1.2; kidney: 7.0 ± 0.5 vs. 27.8 ± 3.1 nmol/g tissue, respectively), catalase (heart: 0.18 ± 0.03 vs. 0.66 ± 0.04; kidney: 0.17 ± 0.03 vs. 0.73 ± 0.03, U/g tissue, respectively) and SOD (heart: 8.01 ± 0.70 vs. 12.3 ± 0.4; kidney: 7.02 ± 0.60 vs. 12.0 ± 0.2, U/g tissue, respectively) showed lower levels in LPS-treated group compared to control; while MDA concentration in LPS group was higher than control (P < 0.05). Histopathological examination in LPS-treated group indicated infiltration of inflammatory cells and more collagen deposition in left ventricle wall and kidney compared to control group. CONCLUSION: We concluded that in clinical conditions with chronic LPS, cardiac and renal fibrosis occurs even in absence of preceding tissue injury due to imbalances in oxidative stress.

Effect of Angiotensin-converting Enzyme Inhibitor on Cardiac Fibrosis and Oxidative Stress Status in Lipopolysaccharide-induced Inflammation Model in Rats.

BACKGROUND: Renin-angiotensin (Ang)-aldosterone system not only plays a key role in the regulation of circulatory homeostasis, but also it acts as a powerful pro-inflammatory mediator. The aim of this study was to evaluate the effect of captopril (Cap), a known Ang-converting enzyme inhibitor, on inflammation-induced cardiac fibrosis, and heart oxidative stress status in lipopolysaccharide (LPS)-induced inflammation in male rats. METHODS: Fifty male rats were randomly divided into five groups control, LPS (1 mg/kg/day), LPS + Cap 10 mg/kg, LPS + Cap 50 mg/kg and LPS + Cap 100 mg/kg. After 2 weeks, blood samples were taken, and hearts were harvested for evaluation of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and nitric oxide metabolite in serum and tissue hemogenate, histopathology (hematoxylin and eosin and Masson's trichrome) and oxidative stress status. RESULTS: Serum IL-6 and TNF-α concentration were higher in LPS group compared to control and Cap reduced them, significantly. Heart TNF-α and IL-6 contents in LPS group were significantly higher than control (P < 0.05). The administration of Cap significantly decreased inflammatory markers level to control (P < 0.05). The higher levels of malondialdehyde and lower antioxidative markers (total thiol, superoxide dismutase, and catalase) in the heart were observed in LPS group and treatment by Cap improved them, dose-dependently. Histopathological study revealed cardiac fibrosis and more collagen content in LPS group which significantly improved by Cap treatment. CONCLUSIONS: Treatment by Cap reduced cardiac fibrosis possibly through improving oxidative stress status, and it can be considered to increase cardiac compliance in this condition.

The effect of Nigella sativa on inflammation-induced myocardial fibrosis in male rats.

Nigella sativa (NS) (Ranunculaceae) used as a protective and therapeutic traditional medicine. This study evaluates the effect of NS on inflammation-induced myocardial fibrosis, serum and tissue inflammatory markers, and oxidative stress status in male rats. Fifty male Wistar rats were divided into five groups: (1) control; (2) lipopolysaccharide (LPS), 1 mg/kg/day; (3) LPS + NS (hydroalcoholic extract), 100 mg/kg/day; (4) LPS + NS, 200 mg/kg/day; (5) LPS + NS, 400 mg/kg/day (n = 10 in each group). The duration of LPS administration was two weeks. At the end of the experiment, blood samples were taken and ventricles were homogenized and stained for histological evaluation. Serum nitrite levels were lower in LPS group than the control group (22.98 ± 1.03 vs 28.5 ± 0.93 µmol/L), in which they were significantly increased by NS treatment (P < 0.05). Higher levels of heart interlukine-6 (IL-6) and tumor necrosis factor-α (TNF-α) were observed in LPS group compared to the controls (IL-6: 6805 ± 656 vs 4733 ± 691 pg/mL; TNF-α: 6504 ± 501 vs 5309 ± 452 pg/mL), in which they were reduced by NS 400 mg/kg compared to LPS groups (P < 0.05). A significant increment of malondialdehyde and reduction in heart total thiol, superoxide dismutase and catalase concentrations were observed in LPS group (p < 0.05) which significantly restored with treatment by three doses of NS. Histopathological studies showed higher inflammatory cell infiltrates, cardiac fibrosis, and collagen deposition in LPS group, which were reduced by the administration of NS. Treatment by NS reduced myocardial fibrosis in inflammation-induced fibrosis, possibly through improving oxidative/anti-oxidative balance.

Loss of heterozygosity and microsatellite instability as predictive markers among Iranian esophageal cancer patients.

OBJECTIVES: Variation in microsatellite sequences that are dispersed in the genome has been linked to a deficiency in cellular mismatch repair system and defects in several genes of this system are involved in carcinogenesis. Our aim in this study was to illustrate microsatellite DNA alteration in esophageal cancer. MATERIALS AND METHODS: DNA was extracted from formalin fixed paraffin embedded (FFPE) tissues from surgical and matched margin-normal samples. Microsatellite instability (MSI) and loss of heterozygosity (LOH) were studied in 50 cases of esophageal squamous cell carcinoma (ESCC) by amplifying six microsatellite markers: D13S260 (13q12.3), D13S267 (13q12.3), D9S171 (9p21), D2S123 (2p), D5S2501 (5q21) and TP53 (17p13.1) analyzed on 6% denaturing polyacrylamide gel electrophoresis. RESULTS: Statistical analysis indicated a near significant reverse correlation between grade and LOH (P= 0.068, correlation coefficient= -0.272). Specifically, increased LOH in tumor DNA has a significant correlation with increased differentiation from poorly differentiated to well differentiated tumors (P= 0.002 and P= 0.016 respectively). In addition, higher number of chromosomal loci with LOH showed a reverse correlation with lymph node metastasis (P= 0.026, correlation coefficient= -0.485). Furthermore, there was a positive correlation between addiction and MSI (P= 0.026, correlation coefficient= 0.465). CONCLUSION: Microsatellite DNA alterations may be a prognostic tool for detection and the evolution of prognosis in patients with SCC of esophagus. It can be concluded that regional lymph node metastasis would be less likely with increased heterozygote loci and addiction with any of opium, cigarette, water pipe or alcohol can be a susceptibility factor(s) for MSI.

Effects of pomegranate seed oil on oxidative stress markers, serum biochemical parameters and pathological findings in kidney and heart of streptozotocin-induced diabetic rats.

PURPOSE: Oxidative stress due to hyperglycemia is a major cause of diabetes complications. The aim of this study was to evaluate the effects of pomegranate seed oil (PSO) on serum biochemical parameters, cardiomyopathy and nephropathy induced by diabetes mellitus. METHOD: W/A adult rats were divided into four groups (12 each): group 1, received saline (1 mL/kg), group 2, received streptozotocin (STZ, 65 mg/kg, a single dose as i.p.), groups 3 and 4, received STZ + PSO (0.4 and 0.8 mL/kg, daily by gavage, respectively). After three weeks, six rats of each group and one week later the remaining animals were anesthetized, blood samples were taken for measuring serum biochemical parameters. Sections of heart and kidneys were used for histopathological studies and the remaining tissues were homogenized for measuring malondialdehyde (MDA) and total sulfhydryl groups. RESULTS: Significant elevation of serum creatinine and urea, LDL, triglyceride, glucose levels as well as urine markers, MDA levels in tissue homogenates and a significant decrease in total thiol content and serum HDL were observed in STZ-treated group as compared with control group. PSO treatment resulted in a significant decrease in tissue MDA content, serum creatinine and urea levels as well as urine markers as compared with STZ-treated group. Lipid profile was ameliorated with PSO treatment. PSO also significantly reversed STZ-induced depletion in thiol content and histological abnormality. Effect of PSO was more specific at 28th than 21th days of study. CONCLUSION: The results showed that PSO has a protective effect against diabetes complications in rats.

Protective effect of pomegranate seed oil against cisplatin-induced nephrotoxicity in rat.

PURPOSE: Clinical use of cisplatin is limited by its nephrotoxicity. Cisplatin-induced nephrotoxicity is associated with an increase in oxidative stress, leading ultimately to kidney dysfunction. The aim of this study was to investigate the effect of pomegranate seed oil against nephrotoxicity induced by cisplatin in adult rats. METHODS: Animals were divided into four groups. Group I received corn oil (1 mL/kg). Group II received cisplatin (8 mg/kg). Group III and IV received pomegranate seed oil (PSO) 0.4 mL/kg and 0.8 mL/kg one hour before cisplatin injection for 3 days, respectively. Blood samples were collected by cardiac puncture and used for measuring urea and creatinine concentration. Twenty-hour urine samples were collected to measure protein and glucose concentration. The right kidney fixed in formalin for histological examination and the left kidney was homogenized for measurement of malondialdehyde and total sulfhydryl groups. RESULTS: A significant elevation of serum creatinine, urea, urinary glucose, protein concentrations, and non-significant decrease in total thiol content and increase in MDA level in kidney homogenates were observed in cisplatin-treated rats. Also cisplatin reduced animal's body weight. Mild-to-moderate tubular cell necrosis, hyaline casts, and vascular congestion were observed in group II. PSO pre-treatment significantly decreased urinary protein, glucose, and serum creatinine concentration. PSO also caused a decrease in serum urea, renal MDA, and increase in thiol content, but the level of these parameters were not significant. CONCLUSION: The present results suggest that PSO is an effective agent for the prevention of cisplatin-induced renal dysfunction and oxidative damage in rat.

Protective effect of pomegranate seed oil against cisplatin-induced nephrotoxicity in rat.

PURPOSE: Clinical use of cisplatin is limited by its nephrotoxicity. Cisplatin-induced nephrotoxicity is associated with an increase in oxidative stress, leading ultimately to kidney dysfunction. The aim of this study was to investigate the effect of pomegranate seed oil against nephrotoxicity induced by cisplatin in adult rats. METHODS: Animals were divided into four groups. Group I received corn oil (1 mL/kg). Group II received cisplatin (8 mg/kg). Group III and IV received pomegranate seed oil (PSO) 0.4 mL/kg and 0.8 mL/kg one hour before cisplatin injection for 3 days, respectively. Blood samples were collected by cardiac puncture and used for measuring urea and creatinine concentration. Twenty-hour urine samples were collected to measure protein and glucose concentration. The right kidney fixed in formalin for histological examination and the left kidney was homogenized for measurement of malondialdehyde and total sulfhydryl groups. RESULTS: A significant elevation of serum creatinine, urea, urinary glucose, protein concentrations, and non-significant decrease in total thiol content and increase in MDA level in kidney homogenates were observed in cisplatin-treated rats. Also cisplatin reduced animal's body weight. Mild-to-moderate tubular cell necrosis, hyaline casts, and vascular congestion were observed in group II. PSO pre-treatment significantly decreased urinary protein, glucose, and serum creatinine concentration. PSO also caused a decrease in serum urea, renal MDA, and increase in thiol content, but the level of these parameters were not significant. CONCLUSION: The present results suggest that PSO is an effective agent for the prevention of cisplatin-induced renal dysfunction and oxidative damage in rat.

Protective effect of pomegranate seed oil against mercuric chloride-induced nephrotoxicity in rat.

PURPOSE: Heavy metals such as mercury can induce the generation of free radicals and oxidative stress which are associated with tissue injury. The present study was designed to evaluate the protective effect of pomegranate seed oil against HgCl2-induced nephrotoxicity. METHODS: Twenty-four W/A adult rats were randomly divided into four groups. Group I received corn oil (1 mL/kg). Group II received HgCl2 (5 mg/kg) for 3 days. Group III and IV received PSO 0.4 mL/kg and 0.8 mL/kg, respectively one hour before HgCl2 administration for 3 days. Blood samples were taken by cardiac puncture and used for the measurement of urea and creatinine concentration. Twenty-hour-hour urine samples were collected to measure protein and glucose. The right kidney was fixed in formalin for histological examination and the left kidney was homogenized for measuring malondialdehyde (MDA) and total sulfhydryl groups. RESULTS: Significant elevation of serum creatinine and urea levels as well as urine glucose and protein concentrations, a significant decrease in total thiol content and a significant increase in MDA levels in kidney homogenate samples were observed after administration of HgCl2 as compared with control group. PSO pretreatment resulted in a significant decrease in serum creatinine and urea levels as well as urine glucose and protein concentrations when compared with HgCl2 treated (group II). PSO also significantly reversed the HgCl2-induced depletion in thiol content and elevation in MDA content. Histological studies revealed milder kidney lesions in PSO treated groups (groups III and IV) compared to HgCl2 treated group. CONCLUSION: Our results suggest that PSO has a protective effect against HgCl2-induced nephrotoxicity in rats.

Old-world leishmaniasis presenting as a nasal mass.

Mucosal leishmaniasis is a major problem in Latin America but has been rarely noticed in our region. Although there have been a few reports of mucosal involvement especially in the oral cavity from Southwest Iran, yet none have been presented from the Northeast where Leishmania tropica is the major concern. We report a patient with endonasal leishmaniasis due to L. tropica, an extremely rare entity in immunocompetent patients in our region. He presented with a mass in the left nasal vestibule causing a sense of obstruction on the same side, accompanied by occasional rhinorrhea and mild epistaxis. This case exemplifies the need for considering leishmaniasis as a differential diagnosis for nasal obstruction in this endemic area.

Analysis of clinicopathologic correlations in Iranian patients with lupus nephritis.

OBJECTIVE: To determine clinical features of different histopathological presentations in patients with lupus nephritis (LN). METHODS: Clinical and pathological features of 71 biopsy-proven LN patients were analyzed in a cross-sectional study during 2005-2011. RESULTS: Sixty-five women (91.5%) and six men (8.5%) were studied. The mean Activity Index (AI) and Chronicity Index (CI) were 6.2 ± 3.1 and 1.7 ± 1.5, respectively. The most common histopathologic presentation of kidneys was class IV (52.1%). Patients with more advanced International Society of Nephrology and the Renal Pathology Society (ISN/RPS) classes, had longer disease duration (P = 0.007), higher levels of blood urea nitrogen (P = 0.004) and serum creatinine (P = 0.035). The most frequent active lesion seen in renal biopsies was endocapillary hypercellularity (83.1%) while glomerular sclerosis was the most common chronic lesion (52.1%). Patients with chronic lesions, including glomerular sclerosis (P = 0.032), fibrous crescent (P = 0.001), interstitial fibrosis (P = 0.025) and tubular atrophy (P = 0.049) had higher serum creatinine levels. Hypertension was mainly seen in patients who had interstitial fibrosis and tubular atrophy (P = 0.026, 0.002 respectively). Moreover, subjects with renal failure had been more frequently involved with fibrinoid necrosis/karyorrhexis (P = 0.003), interstitial inflammation (P = 0.009), fibrous crescents (P = 0.041), tubular atrophy (P = 0.008) and interstitial fibrosis (P < 0.001). CONCLUSION: We found that both histopathologic classification (ISN/RPS criteria) and histopathologic grading (US National Institutes of Health activity and chronicity indices) correlate to some clinical manifestations of LN. Considering these correlations may help to determine the patients' clinicopathologic status, prognosis and the need to immediate treatment. Nevertheless, it is necessary to clarify the accuracy of these findings in larger-scale prospective studies.

Evaluation of HER2/neu oncoprotein in serum and tissue samples of women with breast cancer: correlation with clinicopathological parameters.

BACKGROUND: HER2/neu (HER2) is a proto-oncogen of the EGF Receptor family. The assessment of serum HER2 level is useful for predicting the patients' response to chemotherapy or hormonal therapy and selection of proper patients for treatment with Herceptin. We aimed to compare serum HER2 levels with immunohistochemistry in tumoral tissues and investigate correlation between these levels and various prognostic factors. MATERIALS AND METHODS: This cross-sectional study was conducted on 75 patients with breast carcinoma referred to surgical ward of Mashhad Imam Reza's hospital from November 2008 to February 2009. Pre-operative serum samples were collected and stored in -20°C. Surgical samples were investigated for the type of carcinoma, tumor size, lymph node metastasis, stage as well as grade of the tumor. Tissue HER2 over-expression was evaluated by immunohistochemistry (IHC) staining and HER2 levels were studied by ELISA method. Statistical analysis was performed by SPSS software. RESULTS: Serum HER2 cut-off level was 18.4 ng/ml; 46.7% of patients were serum HER2-positive and 43% were IHC positive. There was a high statistical correlation between these two parameters (P=0.018). Statistically, there was no significant correlation between serum HER2 and age, tumor size, stage, grade and metastatic lymph nodes (P>0.05). CONCLUSION: Serum HER2 level assay can be considered as a complementary method besides tissue methods.

Clinicopathological significance of E-cadherin, ß-catenin and p53 expression in gastric adenocarinoma.

BACKGROUND: E-cadherin/catenin complexes exert a role in cell adhesion. ß-catenin is a key player in Wnt signaling pathway in gastric cancer. P53 is a tumor suppressor gene which also regulates apoptosis. We assessed the expression of E-cadherin, ß-catenin and p53 in gastric adenocarcinoma, and their correlations with clinicopathological features. METHODS: Fifty six formalin-fixed, paraffin-embedded archival specimens of gastric adenocarcinoma were randomly included as cases. Adjacent tumor-free gastric mucosa of different premalignant stages was obtained from the cases. Immunohistochemical staining was performed to assess E-cadherin, ß-catenin and p53 expression. RESULTS: All chronic atrophic gastritis and intestinal metaplasia revealed normal membranous staining. Only one patient with dysplasia had abnormal expression of E-cadherin and ß-Catenin. Abnormal E-cadherin, ß-catenin and p53 expression was found in 50%, 48.2% and 76.8% of cancer specimens respectively. Abnormal expression of E-cadherin was significantly correlated with aberrant ß-catenin expression. Abnormal E-cadherin and ß-catenin expression were significantly correlated with depth of tumor invasion and advanced gastric cancer (p < 0.05), lower degree of differentiation and diffused tumor type (p < 0.001). Node metastasis was not influenced by abnormal expression of E-cadherin and ß-catenin. P53 was not associated with clinicopathological variables. CONCLUSIONS: Abnormal expression of the E-cadherin and ß-catenin were associated with each other and influenced by histogenesis of gastric cancer and malignant behavior of tumor but not significant in premalignant lesions. They are more frequent in diffuse type and associated with advanced gastric cancer. P53 alterations are more frequent in the Iranian population compared with others.

Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran.

AIM: To detect tumor-associated DNA changes in stool samples among Iranian patients with colorectal cancer (CRC) compared to healthy individuals using BAT-26, p16 hypermethylation and long DNA markers. METHODS: Stool DNA was isolated from 45 subjects including 25 CRC patients and 20 healthy individuals using a new, fast and easy extraction method. Long DNA associated with tumor was detected using polymerase chain reaction method. Microsatellite studies were performed utilizing denaturating polyacrylamide gel to determine the instability of BAT-26. Methylation status of p16 promoter was analyzed using methylation-specific PCR (MSP). RESULTS: The results showed a significant difference in existence of long DNA (16 in patients vs 1 in controls, P < 0.001) and p16 (5 in patients vs none in controls, P = 0.043) in the stool samples of two groups. Long DNA was detected in 64% of CRC patients; whereas just one of the healthy individuals was positive for Long DNA. p16 methylation was found in 20% of patients and in none of healthy individuals. Instability of BAT-26 was not detected in any of stool samples. CONCLUSION: We could detect colorectal cancer related genetic alterations by analyzing stool DNA with a sensitivity of 64% and 20% and a specificity of 95% and 100% for Long DNA and p16 respectively. A non-invasive molecular stool-based DNA testing can provide a screening strategy in high-risk individuals. However, additional testing on more samples is necessary from Iranian subjects to determine the exact specificity and sensitivity of these markers.