Characterization and comparison of insulinoma tumor model and pancreatic damage caused by the tumor, and identification of possible markers.

Insulinoma is a neuroendocrine tumor. It arises from the uncontrolled proliferation of pancreatic ß cells. In this study, we created an insulinoma tumor model in nude mice. INS-1 cells were injected in two different ways, subcutaneously (S.C.) or intraperitoneally (I.P.). Body weight, tumor weight, and size were measured. ELISA kits were used analyze to Glucose, insulin, and CA19-9 levels in serum, pancreas, and tumor tissues. KCNN4, KCNK1, GLUT2, IR, HSP70, HSF1, and HSP90 levels were analyzed by western blotting of membrane and/or cytosolic fractions of tumor and pancreas tissue. Tumor formation occurred in nude mice, but it did not occur in Wistar albino rats. The tumor has neuroendocrine cell morphology. Insulin and CA19-9 levels increased in pancreas tissue. In tumor tissue, KCNN4 levels were higher in both membrane and cytosolic fractions, while KCNK1 levels were lower in the membrane fraction of the S.C. group. HSP70 levels were also lower in the S.C. group. In pancreas tissue, KCNK1 levels were lower in the membrane fraction of the S.C. and I.P. groups. GLUT2 levels increased in both groups according to the control group, while IR levels decreased in the S.C. group compared to the control group. However, HSF1 levels increased in the I.P. group, while HSP90 decreased in the S.C. group in pancreatic tissues. The S.C. group is a more suitable insulinoma tumor model. KCNN4, KCNK1, and HSP70 proteins may be important biomarkers in the diagnosis and treatment of insulinoma.

The effect of platelet-rich fibrin, platelet-rich plasma, and concentrated growth factor in the repair of full thickness rotator cuff tears.

BACKGROUND: Rotator cuff lesions rank among the prevalent causes of shoulder pain. Combining surgical interventions with growth factors, scaffolds, and stem cell therapies can effectively decrease the likelihood of rotator cuff repair recurrence. Platelet-rich plasma (PRP), platelet-rich fibrin (PRF), and concentrated growth factor (CGF), isolated from blood and rich in growth factors, have a critical role in cell migration, cell proliferation, and angiogenesis during the tissue regeneration process. Investigations have further substantiated the beneficial impact of PRP and PRF on the biomechanical and histologic attributes of the tendon-bone interface. We aimed to investigate the effectiveness of CGF compared with PRF and PRP in the repair of rotator cuff lesions as a new treatment strategy. METHODS: Incision was performed on both shoulder regions of 21 adult rabbits. After 8 weeks, both shoulders of the rabbits were repaired by suturing. PRF and CGF were administered to 2 separate groups along with the repair. Tissues were collected for biomechanical measurements and histologic evaluations. RESULTS: Histologically, CGF, PRF, and PRP showed similar results to the healthy control group. The level of improvement was significant in the PRF and PRP groups. In the PRF group, the distribution of Ki67 (+), CD31 (+), and CD34 (+) cells was determined intensely in the tendon-bone junction regions. Apoptotic cells increased significantly in the repair group compared with the healthy group, whereas fewer apoptotic cells were found in the PRF-, PRP-, and CGF-applied groups. In the biomechanical results, no statistical difference was recorded among the groups. CONCLUSION: The use of PRF, PRP, and CGF in rotator cuff repair shows promise in shortening the treatment period and preventing the recurrence of rotator cuff lesions.

Kaempferol treatment ameliorates memory impairments in STZ­induced neurodegeneration by acting on reelin signaling.

Many treatment initiatives, like herbal products and their active ingredients, aim to alleviate neurodegeneration to increase cognitive functions. Kaempferol may be a candidate molecule for treating neurodegeneration because of its antioxidant effects. In the present study, we examined the molecular changes associated with kaempferol's memory­enhancing effects on streptozotocin (STZ)­induced neurodegeneration. After intracerebroventricular STZ injection in Long­Evans male rats, intraperitoneal kaempferol was administered for 12 days. The Morris water maze (MWM) was used to measure learning and memory performance in the rats, and proteins related to memory formation were investigated in the hippocampi with western blotting. Kaempferol improved learning performance and memory decline in STZ­treated rats. At the molecular level, STZ­induced neurodegeneration resulted in a decrease in the expression of GAD67, reelin, and phosphorylated­NMDAR. However, kaempferol treatment ameliorated these changes by enhancing their levels similar to the controls. While neither STZ injection nor kaempferol treatment produced any significant change in phosphorylated­CAMKII levels, they increased the expression of klotho and prealbumin. These results show that kaempferol has positive effects on memory loss, affecting synaptic plasticity by ameliorating both the levels and activity of memory­relevant molecules through reelin signaling. In summary, this study provides a guide to future studies by examining in detail the healing effect of kaempferol as a candidate molecule in the treatment of neurodegeneration, such as that observed in Alzheimer's disease.

Examination of antimicrobial effect of fluoxetine in experimental sepsis model: An in vivo study.

Since most infectious diseases can develop into sepsis, it is still a major medical problem. Some in-vivo studies showed promising properties of fluoxetine in the treatment of infections. This study aims the antimicrobial effect of fluoxetine on the inflammatory process used in the treatment of sepsis-modeled rats. Besides, to investigate the efficacy of fluoxetine on modifying the antibiotic effect of imipenem in the inflammatory response. An experimental sepsis model was divided into negative control, positive control, fluoxetine 5 mg/kg, imipenem 60 mg/kg, and combined (fluoxetine; imipenem). Procalcitonin (PCT), high-sensitivity C-reactive protein (hs-CRP), lactate, myeloperoxidase activity (MPO), the inflammation markers interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-alfa (TNF-α), and monocyte chemoattractant protein-1 (MCP-1) levels were measured by enzyme-linked immunosorbent assay method. Oxidative stress markers, total oxidant status (TOS), total antioxidant status (TAS), total thiol (TT), and native thiol (NT) were measured using photometric methods. Oxidative stress index (OSI) was calculated according to TAS and TOS levels. The statistical analysis was performed by Statistical Package for Social Sciences version 22.0. After treatment with fluoxetine, imipenem, and combined groups, IL-1ß, IL-6, TNF-α, MPO activity, MCP-1, hs-CRP, PCT, lactate, and the oxidative stress markers OSI, and disulfide levels were decreased (p < 0.05). The TT, NT, and TAS levels significantly statistically increased (p < 0.05). This research demonstrates that fluoxetine has effects as anti-inflammatory and antioxidant, and the combined treatment with antibioticum imipenem indicates positive synergistic effects in the experimental sepsis model.

Targeting breast cancer using pirarubicin-loaded vasoactive intestinal peptide grafted sterically stabilized micelles.

In this study the chemotherapeutic agent Pirarubicin (PRB) which is known for its serious side effects was actively targeted to the breast cancer cells by uploading it to the biocompatible and biodegradable Sterically Stabilized Micelles (SSMs) made of 1,2- Distearoyl- sn- glycero­3- phosphoethanolamine- N- methoxy­ polyethylene glycol 2000 (DSPE-PEG2000) to enhance efficacy and reduce toxicity. Vasoactive intestinal peptide (VIP), the receptors of which are overexpressed on the breast cancer cells, was grafted on the surface of the micelles. To the best of our knowledge this is the first report on active targeting of PRB to tumor site. For this purpose, PRB loaded VIP grafted SSMs (PRB-SSM-VIP) were synthesized and characterized. The in vitro efficiency of PRB-SSM-VIP along with SSM and free PRB was investigated on the MCF-7 breast cancer cells and the in vivo effects were studied on the 4T1 breast cancer bearing nude mice. Solubilizing 300 µg of PRB using 2.81 mg of DSPE-PEG2000 resulted in obtaining monodispersed particles of 12.16 ± 2.7 nm with slow drug release profile. Incorporation of PRB within the hydrophobic DSPE core of SSM was confirmed using differential scanning calorimetry (DSC) and the spherical shape of the synthesized particles was demonstrated using atomic force microscope (AFM). Both in vitro and in vivo studies showed significantly higher activity of PRB-SSM-VIP compared to free PRB. In vivo imaging showed successful accumulation of PRB-SSM-VIP at the tumor site and 98.8% tumor eradication was obtained with no signs of side effects. Current study suggests that SSM-VIP could be used as new drug delivery system for targeting PRB to the breast cancer cells.

Premature Myocardial Infarction: Genetic Variations in SIRT1 Affect Disease Susceptibility.

OBJECTIVES: Premature myocardial infarction (PMI) is an uncommon disease, and its incidence varies between 2% and 10%, rising, depending on genetic susceptibility under the influence of lifestyle. The purpose of this study was to investigate the association between SIRT1 single nucleotide polymorphisms (SNPs), SIRT1, and eNOS (endothelial nitric oxide synthase) protein expressions, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) in young patients with premature ST-elevation myocardial infarction (STEMI). METHODS: Genotyping of the three single-nucleotide polymorphisms (rs7895833 A > G in the promoter region, rs7069102 C > G in intron 4, and rs2273773 C > T in exon 5) in SIRT1 gene was performed in 108 consecutive patients (87.0% were men with a mean age of 40.74 ± 3.82 years) suffering from ST-elevation myocardial infarction at the age of ≤45 and 91 control subjects. RESULTS: The risk for myocardial infarction was increased by 2.31 times in carriers of CC or CG genotypes. SIRT1 protein levels were enhanced and endothelial nitric oxide synthase levels were diminished in ST-elevation myocardial infarction patients regardless of the underlying gene variant. There was no correlation between SIRT1 expression and the amount of endothelial nitric oxide synthase, total antioxidant status, total oxidant status, and oxidative stress index levels in patients and in the control group either. CONCLUSIONS: SIRT1 single-nucleotide polymorphisms were associated with premature myocardial infarction, which affected the SIRT1 and endothelial nitric oxide synthase protein expression, irrespective of the underlying SIRT1 genotype.

The beneficial effects of subcutaneous Allium cepa injection on random flap survival in rats.

Random flaps are frequently used in the practice of reconstructive surgery. The aim of this experimental study was to investigate the effects of Allium cepa on random flap survival in rats. Fourteen Wistar rats were used for this experimental study. The subjects were divided into experiment and control groups. Rats in the experiment group received daily injections of A. cepa extract for 7 d before the elevation of the flaps. Following the treatment period, elevation and reinsertion of the dorsal flaps were performed. Necrotic and total flaps areas were marked and calculated 7 d after the flap elevation. Histological examinations and microangiography were performed to evaluate the results. The mean value of the proportion of necrotic flap areas to the total flap area was 25.06 and 50.6% in the A. cepa and control group, respectively (p = .0079). In the histological examination, number of vessels identified in the dermis were 23.75 ± 0.7 and 33.75 ± 9 in the A. cepa and control group, respectively (p = .7457). In angiographic images, vessels formations were more noticeable in the A. cepa group. We conclude that preoperative subcutaneous A. cepa injection increases dorsal flap survival in rats.

Preoperative subcutaneous sildenafil injection increases random flap survival in rats.

PURPOSE: To investigate the effect of subcutaneous sildenafil on random flap survival. METHODS: Fourteen Wistar rats, which were divided in to two groups, were used for this experimental study. Rats in the sildenafil group received subcutaneous sildenafil injections daily for seven days before flap elevation. At the end of the treatment period, 9x3 cm dorsal skin flaps were elevated and reinserted back into their place in all of the animals. Necrotic and whole flaps areas were recorded on graph papers. Seven days after the flap elevation samples for histological examination were taken and angiographies were performed to visualize the flap vascularization. RESULTS: The calculated average percentage of necrotic flap areas were 18.29% and 42.26% in the sildenafil and control group respectively.(p=0.0233). In selected angiography images, vessels were found to be more prominent in the sildenafil group. The average number of capillary formations under light microscopy was higher in the sildenafil group (p= 0.0286). CONCLUSION: The subdermal high dose sildenafil has a positive effect on flap survival.

Preoperative subcutaneous sildenafil injection increases random flap survival in rats

Abstract Purpose: To investigate the effect of subcutaneous sildenafil on random flap survival. Methods: Fourteen Wistar rats, which were divided in to two groups, were used for this experimental study. Rats in the sildenafil group received subcutaneous sildenafil injections daily for seven days before flap elevation. At the end of the treatment period, 9x3 cm dorsal skin flaps were elevated and reinserted back into their place in all of the animals. Necrotic and whole flaps areas were recorded on graph papers. Seven days after the flap elevation samples for histological examination were taken and angiographies were performed to visualize the flap vascularization. Results: The calculated average percentage of necrotic flap areas were 18.29% and 42.26% in the sildenafil and control group respectively.(p=0.0233). In selected angiography images, vessels were found to be more prominent in the sildenafil group. The average number of capillary formations under light microscopy was higher in the sildenafil group (p= 0.0286). Conclusion: The subdermal high dose sildenafil has a positive effect on flap survival.

The effects of lipoic acid on redox status in brain regions and systemic circulation in streptozotocin-induced sporadic Alzheimer's disease model.

While the deterioration of insulin-glucose metabolism (IGM), impaired redox homeostasis (IRH), ß-amyloid accumulation was reported in Sporadic Alzheimer's Disease (SAD) model, aforementioned factors related to lipoic acid administration and anthropometric indexes (AIs) are not yet studied with integrative approach. ß-amyloid accumulation, redox homeostasis biomarkers and AIs are investigated in SAD model. Streptozotocin-induced inhibition of insulin-signaling cascade but not GLUT-2 and GLUT-3 transporters takes a role in ß-amyloid accumulation. Inhibition types are related to IRH in cortex, hippocampus and systemic circulation. Lipoic acid (LA) shows both antioxidant and prooxidant effect according to the anatomical location. LA administration also leads to improved AIs during GLUT-2 inhibition and cortical redox status in GLUT-3 inhibited group. Optimal LA action could be possible if its redox behavior is balanced to antioxidant effect. Diagnostic usage of systemic IRH parameters as biomarkers and their possible correlations with deteriorated IGM should be investigated. Graphical abstract ᅟ.